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1. Compound ID: 135
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-4)-b-D-ManpNAc3NAmA-(1-4)-b-D-ManpNAc3NAcA-(1-3)-a-D-FucpNAc-(1- |
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Structure type: polymer chemical repeating unit
Aglycon: core
Compound class: O-polysaccharide
Contained glycoepitopes: IEDB_2275410
The structure is contained in the following publication(s):
- Article ID: 27
Burrows LL, Charter DF, Lam JS "Molecular characterization of the Pseudomonas aeruginosa serotype O5 (PAO1) B-band lipopolysaccharide gene cluster" -
Molecular Microbiology 22 (1996) 481-495
Pseudomonas aeruginosa co-expresses A-band lipopolysaccharide (LPS), a homopolymer of rhamnose, and B-band LPS, a heteropolymer with a repeating unit of 2-5 sugars which is the serotype-specific antigen. The gene clusters for A- and B-band biosynthesis in P. aeruginosa O5 (strain PAO1) have been cloned previously. Here we report the DNA sequence and molecular analysis of the B-band O-antigen biosynthetic cluster. Sixteen open reading frames (ORFs) thought to be involved in synthesis of the O5 O antigen were identified, including wzz (rol), wzy (rfc), and wbpA-wbpN. A further 3 ORFs not thought to be involved with LPS synthesis were identified (hisH, hisF, and uvrB). Most of the wbp genes are found only in serotypes O2, O5, O16, O18, and O20, which form a chemically and structurally related O-antigen serogroup. In contrast, wbpM and wbpN are common to all 20 serotypes of P. aeruginosa. Although wbpM is not serogroup-specific, knockout mutations confirmed it is necessary for O5 O-antigen biosynthesis. A novel insertion sequences, IS 1209, is present at the junction between the serogroup-specific and non-specific regions. We have predicted the functions of the proteins encoded in the wbp cluster based on their homologies to those in the databases, and provide a proposed pathway of P. aeruginosa O5 O-antigen biosynthesis
Lipopolysaccharide, biosynthesis, LPS, gene, genetics, characterization, serotype, B-band, Pseudomonas, Pseudomonas aeruginosa, molecular, cluster, gene cluster, O-polysaccharide
NCBI PubMed ID: 8939432Journal NLM ID: 8712028Publisher: Blackwell Publishing
Correspondence: jlam@micro.uoguelph.ca
Institutions: Department of Microbiology, University of Guelph, Ontario, Canada
Methods: DNA sequencing
- Article ID: 1168
Sadovskaya I, Brisson J, Thibault P, Richards JC, Lam JS, Altman E "Structural characterization of the outer core and the O-chain linkage region of lipopolysaccharide from Pseudomonas aeruginosa serotype O5" -
European Journal of Biochemistry 267 (2000) 1640-1650
The point of attachment of the O-chain in the outer core region of Pseudomonas aeruginosa serotype O5 lipopolysaccharide (LPS) was determined following a detailed analysis of the extended core oligosaccharide, containing one trisaccharide O-chain repeating unit, present in both the wild-type strain PAO1 and O-chain deficient mutant strains AK1401 and PAO-rfc. The structure of the extended core oligosaccharide was determined by various mass spectrometric methods as well as one-dimensional and two-dimensional NMR spectroscopy. Furthermore, the one-dimensional analogues of NOESY and TOCSY experiments were applied to confirm the structure of the outer core region in the O-chain polysaccharide. In both the extended core oligosaccharide and the core of the smooth LPS, a loss of one of the β-glucosyl residues and the translocation of the α-rhamnosyl residue, followed by the attachment of the first O-chain repeating unit was observed. This process is complicated and could involve two distinct rhamnosyltransferases, one with α-1,6-linkage specificity and another with α-1,3-linkage specificity. It is also plausible that an α-1,3 rhamnosyltransferase facilitates the addition of the 'new' α-rhamnosyl residue that will act as a receptor for the attachment of the single O-antigen repeating unit in the LPS of the semi-rough mutant. The 2-amino-2-deoxy-fucosyl residue of the first O-chain repeating unit directly attached to the core was found to have a β-anomeric configuration instead of an α configuration, characteristic for this residue as a component of the O-chain polysaccharide. The results of this study provide the first example of the mechanistic implications of the structure of the outer core region in a fully assembled O-chain containing LPS, differing from the O-chain deficient rough LPS
Lipopolysaccharide, LPS, oligosaccharide, core, strain, structural, characterization, serotype, analysis, Pseudomonas, Pseudomonas aeruginosa, core oligosaccharide, wild type, mutant, linkage, core region, region, elucidation, O-chain, O-chain linkage region
NCBI PubMed ID: 10712594Journal NLM ID: 0107600Publisher: Oxford, UK: Blackwell Science Ltd. on behalf of the Federation of European Biochemical Societies
Correspondence: eleonora.altman@nrc.ca
Institutions: Institute for Biological Sciences, National Research Council of Canada, Ottawa, Ontario K1A 0R6, Canada, Department of Microbiology, College of Biological Sciences, University of Guelph, Ontario N1G 2W1, Canada
Methods: 13C NMR, 1H NMR, NMR-2D, ESI-MS
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2. Compound ID: 159
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a-D-GlcpN-(1-7)-+
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a-D-GlcpA-(1-2)-L-gro-a-D-manHepp-(1-3)-+
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a-D-GalpNA-(1-6)-+ |
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L-gro-a-D-manHepp-(1-4)-+ | | EtN-(1--P--4)--+
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a-D-GlcpNAc-(1-4)-b-D-ManpNAc3NAcA-(1-3)-b-L-FucpNAc4NMe-(1-6)-a-D-GlcpN-(1-4)-b-D-Glcp-(1-4)-L-gro-a-D-manHepp-(1-5)-a-Kdop-(2--/lipid A/ |
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Structure type: oligomer
Aglycon: lipid A
Compound class: LPS
Contained glycoepitopes: IEDB_115136,IEDB_120354,IEDB_123890,IEDB_130650,IEDB_137340,IEDB_137777,IEDB_140087,IEDB_140088,IEDB_140090,IEDB_140630,IEDB_141807,IEDB_142488,IEDB_146664,IEDB_151531,IEDB_2189047,IEDB_2275071,IEDB_2275073,IEDB_2275074,IEDB_983931,SB_192
The structure is contained in the following publication(s):
- Article ID: 36
Caroff M, Brisson J, Martin A, Karibian D "Structure of the Bordetella pertussis 1414 endotoxin" -
FEBS Letters 477 (2000) 8-14
The endotoxin (lipopolysaccharide) of Bordetella pertussis, the agent of whooping cough, consists of a lipid A linked to a highly branched dodecasaccharide containing several acid and amino sugars. The elucidation of the polysaccharide structure was accomplished by first analyzing the structures of fragments obtained by hydrolysis and nitrous deamination and then piecing the fragments together. The fine structure of the antigenic distal pentasaccharide, presented here, was determined by chemical analyses as well as by high-resolution nuclear magnetic resonance and mass spectrometry. The complete structure was reconstituted and confirmed by matrix-assisted laser desorption/ionization mass spectrometry. The following structure was derived from the combined experimental data:The detailed structure combined with previously reported serological data now allows the synthesis of its epitopes for potential vaccines
structure, Bordetella, Bordetella pertussis, endotoxin, pertussis
NCBI PubMed ID: 10899302Publication DOI: 10.1016/s0014-57930001720-8Journal NLM ID: 0155157Publisher: Elsevier
Correspondence: martine.caroff@bbmpc.u-psud.fr
Institutions: Equipe Endotoxines, UMR 8619 du Centre National de la Recherche Scientifique, Biochimie, Universite de Paris- Sud, F-91405, Orsay, France
Methods: 13C NMR, 1H NMR, NMR-2D, conformation analysis, GC, b-elimination, Smith degradation, MALDI-MS
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3. Compound ID: 171
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a-D-GlcpN-(1-7)-+
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a-D-GlcpA-(1-2)-L-gro-a-D-manHepp-(1-3)-+
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a-D-GalpNA-(1-6)-+ |
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L-gro-a-D-manHepp-(1-4)-+ | | EtN-(1-0)-?%P-4)-+
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a-D-GlcpNAc-(1-4)-b-D-ManpNAc3NAcA-(1-3)-b-L-FucpNAc4NMe-(1-6)-a-D-GlcpN-(1-4)-b-D-Glcp-(1-4)-L-gro-a-D-manHepp-(1-5)-a-Kdop-(2--/lipid A/ |
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Structure type: oligomer
Aglycon: lipid A
Compound class: LPS
Contained glycoepitopes: IEDB_115136,IEDB_120354,IEDB_123890,IEDB_130650,IEDB_137340,IEDB_137777,IEDB_140087,IEDB_140088,IEDB_140090,IEDB_140630,IEDB_141807,IEDB_142488,IEDB_146664,IEDB_151531,IEDB_2189047,IEDB_2275071,IEDB_2275073,IEDB_2275074,IEDB_983931,SB_192
The structure is contained in the following publication(s):
- Article ID: 37
Caroff M, Karibian D "Structure of bacterial lipopolysaccharides" -
Carbohydrate Research 338(23) (2003) 2431-2447
Bacterial lipopolysaccharides are the major components of the outer surface of Gram-negative bacteria They are often of interest in medicine for their immunomodulatory properties. In small amounts they can be beneficial, but in larger amounts they may cause endotoxic shock. Although they share a common architecture, their structural details exert a strong influence on their activity. These molecules comprise: a lipid moiety, called lipid A, which is considered to be the endotoxic component, a glycosidic part consisting of a core of approximately 10 monosaccharides and, in 'smooth-type' lipopolysaccharides, a third region, named O-chain, consisting of repetitive subunits of one to eight monosaccharides responsible for much of the immunospecificity of the bacterial cell.
Lipopolysaccharide, structure, core, lipid A, endotoxin, O-chains
NCBI PubMed ID: 14670707Publication DOI: 10.1016/j.carres.2003.07.010Journal NLM ID: 0043535Publisher: Elsevier
Correspondence: martine.carloff@bbmpc.u-psud.fr
Institutions: Equipe Endotoxines, UMR 8619 du Centre National de la Recherche Scientifique, IBBMC, Université de Paris-Sud, F-Orsay, France
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4. Compound ID: 901
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-4)-b-D-ManpNAc3NAmA-(1-4)-b-D-ManpNAc3NAcA-(1-3)-b-D-FucpNAc-(1- |
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Structure type: polymer chemical repeating unit
Compound class: O-polysaccharide, O-antigen
The structure is contained in the following publication(s):
- Article ID: 254
Hatano K, Pier GB "Complex serology and immune response of mice to variant high-molecular-weight O polysaccharide isolated from Pseudomonas aeruginosa serogroup O2 strains" -
Infection and Immunity 66(8) (1998) 3719-3726
The O antigen of the Pseudomonas aeruginosa lipopolysaccharide is the optimal target for protective antibodies, but the unusual and complex nature of their sugar substituents has made it difficult to define the range of these structures needed in an effective vaccine. Most clinical isolates of P. aeruginosa can be classified into 10 O-antigen serogroups, but slight chemical differences among O polysaccharides within a serogroup give rise to subtype epitopes. These epitopes could impact the reactivity of O-antigen-specific antibodies, as well as the susceptibility of a target strain to protective, opsonic antibodies. To define parameters of serogroup and subtype-epitope immunogenicity, antigenicity, and surface expression on P. aeruginosa cells, we prepared high-molecular-weight O-polysaccharide vaccines from strains of P. aeruginosa serogroup O2, for which eight structurally variant O antigens expressing six defined subtype epitopes (O2a to O2f) have been identified. A complex pattern of immune responses to these antigens was observed following vaccination of mice. The high-molecular-weight O polysaccharides were generally more immunogenic at low doses (1 and 10 microg) than at a high dose (50 microg) and usually elicited antibodies that opsonized the homologous strain for phagocytic killing. Some of the individual polysaccharides elicited cross-opsonic antibodies to a variable number of strains that express all of the defined serogroup O2 subtype epitopes. Combination into one vaccine of two antigens that individually elicited cross-reactive opsonic antibodies to most members of the O2 serogroup inhibited, instead of enhanced, the production of antibodies broadly reactive with most serogroup O2 subtype strains. Thus, immune responses to P. aeruginosa O antigens may be restricted to a limited range of epitopes on structurally complex O antigens, and combining multiple related antigens into a single vaccine formulation may inhibit the production of those antibodies best able to protect against most P. aeruginosa strains within a given O-antigen serogroup.
Lipopolysaccharide, Pseudomonas, Pseudomonas aeruginosa, O-polysaccharide, serology, serogroup, immune response
NCBI PubMed ID: 9673254Journal NLM ID: 0246127Publisher: American Society for Microbiology
Correspondence: gpier@channing.harvard.edu
Institutions: Charming Laboratory, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115-5804., Charming Laboratory, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School,Boston, Massachusetts 02115-5804.
- Article ID: 337
Newton GJ, Daniels C, Burrows LL, Kropinski AM, Clarke AJ, Lam JS "Three-component-mediated serotype conversion in Pseudomonas aeruginosa by bacteriophage D3" -
Molecular Microbiology 39(5) (2001) 1237-1247
Bacteriophage D3 is capable of lysogenizing Pseudomonas aeruginosa PAO1 (serotype O5), converting the O-antigen from O5 to O16 and O-acetylating the N-acetylfucosamine moiety. To investigate the mechanism of lysogenic conversion, a 3.6 kb fragment from the D3 genome was isolated capable of mediating serotypic conversion identical to the D3 lysogen strain (AK1380). The PAO1 transformants containing this 3.6 kb of D3 DNA exhibited identical lipopolysaccharide (LPS) banding patterns to serotype O16 in silver-stained SDS-PAGE gels and displayed reactivity to an antibody specific for O-acetyl groups. Further analysis led to the identification of three open reading frames (ORFs) required for serotype conversion: an α-polymerase inhibitor (iap); an O-acetylase (oac); and a β-polymerase (wzyβ). The α-polymerase inhibitor (Iap) is capable of inhibiting the assembly of the serotype-specific O5 B-band LPS and allows the phage-encoded β-polymerase (Wzyβ) to form new β-linked B-band LPS. The D3 phage also alters the LPS by the addition of O-acetyl groups to the FucNAc residue in the O-antigen repeat unit by the action of the D3 O-acetylase (Oac). These three components form a simple yet elegant system by which bacteriophage D3 is capable of altering the surface of P. aeruginosa PAO1
serotype, Pseudomonas, Pseudomonas aeruginosa, bacteriophage, conversion, serotype conversion
NCBI PubMed ID: 11251840Journal NLM ID: 8712028Publisher: Blackwell Publishing
Correspondence: jlam@uoguelph.ca
Institutions: Department of Microbiology, University of Guelph, Guelph, Ontario, N1G 2W1, Canada, The Centre for Infection and Biomaterials Research, Toronto General Hospital, Toronto, Ontario, M5G 2C4, Canada, Department of Microbiology and Immunology, Queen's University, Kingston, Ontario, K7L 3N6, Canada
Methods: PCR, SDS-PAGE, DNA techniques, Western blotting, quantification of O-acetylation
- Article ID: 1412
de Kievit TR, Staples T, Lam JS "Pseudomonas aeruginosa rfc genes of serotypes O2 and O5 could complement O-polymerase-deficient semi-rough mutants of either serotype" -
FEMS Microbiology Reviews 147(2) (1997) 251-257
Using a gene-replacement strategy and a mutated copy of the Pseudomonas aeruginosa O5 rfc gene, we were able to generate a rfc mutant in P. aeruginosa serotype O2. This mutant, which exhibits the semi-rough (SR) LPS phenotype, was used to isolate the O2 rfc gene. Mobilization of the O2 and O5 rfc genes into SR mutants of the heterologous serotype resulted in 'cross-polymerization' of O-repeat units, indicating that the genes are functionally exchangeable. Analysis of the nucleotide sequence of the rfc genes revealed that the two Rfc proteins are identical. The results of this study have enabled us to propose the linkage catalyzed by the O5 O-polymerase enzyme.
Lipopolysaccharide, LPS, gene, serotype, Pseudomonas, Pseudomonas aeruginosa, mutant, mutants, Serotypes, O-antigen polymerase, rfc, complement, semirough
NCBI PubMed ID: 9119201Journal NLM ID: 8902526Publisher: Oxford University Press
Correspondence: jlam@micro.uoguelph.ca
Institutions: Department of Microbiology, University of Guelph, Guelph, Canada
- Article ID: 1778
Knirel YA, Kochetkov NK "The structure of lipopolysaccharides of gram-negative bacteria. III. The structure of O-antigens: A review" -
Biochemistry (Moscow) 59(12) (1994) 1325-1383
This review summarizes data on the composition and structure of the O-antigens, the polysaccharide chains of the outer-membrane lipopolysaccharides (LPS) of Gram-negative bacteria defining the immunospecificity of these microbial cells. Special reference is given to some structural features of the O-antigens, such as the presence of unique monosaccharides and noncarbohydrate components, masked regularity, and the occurrence in one microorganism of LPS with structurally different polysaccharide chains. Antigenic relationships between microorganisms belonging to different taxonomic groups are discussed.
structure, O-antigen, chemical composition, bacterial lipopolysaccharides, Salmonella livingstone C1
NCBI PubMed ID: 7533007Journal NLM ID: 0376536Publisher: Nauka/Interperiodica
Institutions: Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia
- Article ID: 2409
Knirel YA, Paramonov NA, Vinogradov EV, Shashkov AS, Dmitriev BA, Kochetkov NK, Kholodkova EV, Stanislavsky ES "Somatic antigens of Pseudomonas aeruginosa. The structure of O-specific polysaccharide chains of lipopolysaccharides of P. aeruginosa O3 (Lányi), O25 (Wokatsch) and Fisher immunotypes 3 and 7" -
European Journal of Biochemistry 167 (1987) 549-561
O-specific polysaccharides, obtained on mild acid degradation of lipopolysacchrides of the serologically related strains Pseudomonas aeruginosa O3 (Lányi classification), O25 (Wokatsch classification) and immunotypes 3 and 7 (Fisher classification), are built up of trisaccharide repeating units involving 2-acetamido-2,6-dideoxy-D-galactose (N-acetyl-D-fucosamine), 2,3-diacetamido-2,3-dideoxy-D-mannuronic acid or 2,3-diacetamido-2,3-dideoxy-L-guluronic acid and 3-acetamidino-2-acetamido-2,3-dideoxy-D-mannuronic acid or 3-acetamidino-2-acetamido-2,3-dideoxy-L-guluronic acid. Lányi O3(a),3d,3f and Wokatsch O25 polysaccharides contain also O-acetyl groups. On the basis of solvolysis with anhydrous hydrogen fluoride, resulting in trisaccharide fragments with N-acetylfucosamine residue at the reducing terminus, chemical modifications of the acetamidino group (alkaline hydrolysis to the acetamido group or reductive deamination to the ethylamino group), as well as analysis by 1H-NMR (including nuclear Overhauser effect experiments) and 13C-NMR spectroscopy, and fast-atom bombardment mass spectrometry, it was concluded that the repeating units of the polysaccharides have the following structures: (Formula: see text) where HexNAcAmA = α-L-GulNAcAmA (approximately 70%) or β-D-ManNacAMA (approximately 30%). Lányi O3(a),3d,3f polysaccharide involves two types of repeating units, which differ from each other only in the configuration at C-5 of the 3-acetamidino-2-acetamido-2,3-dideoxyuronic acid residue. Lányi O3(a),3c,O3a,3d,3e and Fisher immunotypes 3 and 7 polysaccharides contain, together with the major repeating units shown above, a small proportion of units in which the derivative of α-L-guluronic acid is replaced by the corresponding β-D-manno isomer. The data obtained provide the opportunity to substantiate the serological interrelations between these strains of P. aeruginosa by the presence in the O-specific polysaccharides of common monosaccharides or disaccharide fragments. The distinctions between them stem from the presence or absence of the O-acetyl group, a different configuration of the glycosidic linkage of the N-acetylfucosamine residue and/or a different configuration at C-5 of one or both derivatives of diaminouronic acids.
NCBI PubMed ID: 3115777Journal NLM ID: 0107600Publisher: Oxford, UK: Blackwell Science Ltd. on behalf of the Federation of European Biochemical Societies
Institutions: N.D. Zelinsky Institute of Organic Chemistry, Academy of Sciences of the USSR, Moscow, Russia
Methods: 13C NMR, 1H NMR, FAB-MS
- Article ID: 3207
Knirel YA, Bystrova OV, Kocharova NA, Zähringer U, Pier GB "Conserved and variable structural features of the Pseudomonas aeruginosa lipopolysaccharide" -
Journal of Endotoxin Research 12(6) (2006) 324-336
The review is devoted to recent progress in the structural elucidation of the lipopolysaccharide of the bacterium Pseudomonas aeruginosa, including O-antigen biological repeats, core oligosaccharide, and lipid A. Data on biosynthesis, genetics and serology of the lipopolysaccharide isolated from various P. aeruginosa O-serogroups are discussed in relation to the chemical structures.
O-antigen, Pseudomonas aeruginosa, lipid A, core oligosaccharide, lipopolysaccharide structure, serologic activity
NCBI PubMed ID: 17254386Publication DOI: 10.1179/096805106X118906Journal NLM ID: 9433350Publisher: Maney Publishing
Correspondence: knirel@ioc.ac.ru
Institutions: N.D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia, Research Center Borstel, Leibniz-Center for Medicine and Biosciences, Borstel, Germany, Channing Laboratory, Brigham and Women’s Hospital, Harvard Medical School, Boston, Massachusetts, USA
Methods: NMR, chemical methods, MS, serological methods, genetic methods
- Article ID: 3391
Kaluzny K, Abeyrathne PD, Lam JS "Coexistence of Two Distinct Versions of O-Antigen Polymerase, Wzy-Alpha and Wzy-Beta, in Pseudomonas aeruginosa Serogroup O2 and Their Contributions to Cell Surface Diversity" -
Journal of Bacteriology 189(11) (2007) 4141-4152
Assembly of B-band LPS in Pseudomonas aeruginosa follows a Wzy-dependent pathway, requiring the O-antigen polymerase Wzy, and other proteins. The peptide sequences of the wzyα product from strains of serotypes O2, O5, and O16 are identical; but the O-units in O5 are α-glycosidic-linked, while those in O2 and O16 are β-linked. We hypothesized that a derivative of the D3 bacteriophage wzyβ is present in the chromosomes of O2 and O16, and that this gene is responsible for the β-linkage. By a combination of PCR and primer walking, wzyβ of both serotypes have been amplified and cloned. Both are identical, but only share 87.42% sequence identity with their xenolog in D3. A chromosomal knockout mutant of O16 wzyβ was made and it produces rough LPS devoid of B-band O antigen. The cloned wzyβ is capable of complementing the O16 wzyβ mutant, as well as cross-complementing a wzyα knockout mutant. However, in the latter case, the restored O-antigen was β-linked. Using RT-PCR, we showed that wzyα was transcribed in O2 and O16 strains and was functional since both of these genes could complement the wzyα mutant of O5. With the co-existence of wzyα and wzyβ in O2 and O16 and the B-band O polysaccharides in these being β-linked, we hypothesized that iap, an inhibitor of alpha-polymerase gene, must be present in these serotypes. Indeed, through PCR, TOPO cloning and nucleotide sequencing results, we verified the presence of iap in both O2 and O16 serotypes
serotype, Pseudomonas aeruginosa, O-polysaccharide, serogroup, sequencing, O-antigen polymerase, rough LPS
NCBI PubMed ID: 17384183Journal NLM ID: 2985120RPublisher: American Society for Microbiology
Correspondence: jlam@uoguelph.ca
Institutions: University of Guelph, Department of Molecular and Cellular Biology, Guelph, Ontario, Canada N1G 2W1
Methods: serological methods, genetic methods
- Article ID: 3711
King JD, Kocincova D, Westman EL, Lam JS "Lipopolysaccharide biosynthesis in Pseudomonas aeruginosa" -
Innate Immunity 15(5) (2009) 261-312
Pseudomonas aeruginosa causes serious nosocomial infections, and an important virulence factor produced by this organism is lipopolysaccharide (LPS). This review summarizes knowledge about biosynthesis of all three structural domains of LPS - lipid A, core oligosaccharide, and O polysaccharides. In addition, based on similarities with other bacterial species, this review proposes new hypothetical pathways for unstudied steps in the biosynthesis of P. aeruginosa LPS. Lipid A biosynthesis is discussed in relation to Escherichia coli and Salmonella, and the biosyntheses of core sugar precursors and core oligosaccharide are summarised. Pseudomonas aeruginosa attaches a Common Polysaccharide Antigen and O-Specific Antigen polysaccharides to lipid A-core. Both forms of O polysaccharide are discussed with respect to their independent synthesis mechanisms. Recent advances in understanding O-polysaccharide biosynthesis since the last major review on this subject, published nearly a decade ago, are highlighted. Since P. aeruginosa O polysaccharides contain unusual sugars, sugar-nucleotide biosynthesis pathways are reviewed in detail. Knowledge derived from detailed studies in the O5, O6 and O11 serotypes is applied to predict biosynthesis pathways of sugars in poorly-studied serotypes, especially O1, O4, and O13/O14. Although further work is required, a full understanding of LPS biosynthesis in P. aeruginosa is almost within reach.
Lipopolysaccharide, core, O-antigen, Pseudomonas aeruginosa, lipid A
NCBI PubMed ID: 19710102Publication DOI: 10.1177/1753425909106436Journal NLM ID: 101469670Publisher: Sage Publications
Correspondence: jlam@uoguelph.ca
Institutions: Department of Molecular and Cellular Biology, University of Guelph, Guelph, Ontario, Canada
- Article ID: 4062
Lam JS, Taylor VL, Islam ST, Hao Y, Kocincova D "Genetic and Functional Diversity of Pseudomonas aeruginosa Lipopolysaccharide" -
Frontiers in Microbiology 2 (2011) 118
Lipopolysccharide (LPS) is an integral component of the Pseudomonas aeruginosa cell envelope, occupying the outer leaflet of the outer membrane in this Gram-negative opportunistic pathogen. It is important for bacterium-host interactions and has been shown to be a major virulence factor for this organism. Structurally, P. aeruginosa LPS is composed of three domains, namely, lipid A, core oligosaccharide, and the distal O antigen (O-Ag). Most P. aeruginosa strains produce two distinct forms of O-Ag, one a homopolymer of D-rhamnose that is a common polysaccharide antigen (CPA, formerly termed A band), and the other a heteropolymer of three to five distinct (and often unique dideoxy) sugars in its repeat units, known as O-specific antigen (OSA, formerly termed B band). Compositional differences in the O units among the OSA from different strains form the basis of the International Antigenic Typing Scheme for classification via serotyping of different strains of P. aeruginosa. The focus of this review is to provide state-of-the-art knowledge on the genetic and resultant functional diversity of LPS produced by P. aeruginosa. The underlying factors contributing to this diversity will be thoroughly discussed and presented in the context of its contributions to host-pathogen interactions and the control/prevention of infection.
Lipopolysaccharide, biosynthesis, virulence, serotyping, bacteriophage, motility, seroconversion, nucleotide sugars
NCBI PubMed ID: 21687428Publication DOI: 10.3389/fmicb.2011.00118Journal NLM ID: 101548977Publisher: Lausanne: Frontiers Research Foundation
Correspondence: jlam@uoguelph.ca
Institutions: Department of Molecular and Cellular Biology, University of Guelph Guelph, ON, Canada
- Article ID: 4684
Islam ST, Lam JS "Synthesis of bacterial polysaccharides via the Wzx/Wzy-dependent pathway" -
Canadian Journal of Microbiology 60(11) (2014) 697-716
The surfaces of bacteria mediate a multitude of functions in the environment and in an infected host, including adhesion to both biotic and abiotic substrata, motility, immune system interaction and (or) activation, biofilm formation, and cell-cell communication, with many of these features directly influenced by cell-surface glycans. In both Gram-negative and Gram-positive bacteria, the majority of cell-surface polysaccharides are produced via the Wzx/Wzy-dependent assembly pathway; these glycans include heteropolymeric O-antigen, enterobacterial common antigen, exopolysaccharide, spore coat, and capsule in diverse bacteria. The key components of this assembly pathway are the integral inner membrane Wzx flippase, Wzy polymerase, and Wzz chain-length regulator proteins, which until recently have resisted detailed structural and functional characterization. In this review, we have provided a comprehensive synthesis of the latest structural and mechanistic data for each protein, as well as an examination of substrate specificity for each assembly step and complex formation between the constituent proteins. To complement the unprecedented explosion of genomic-sequencing data for bacteria, we have also highlighted both classical and state-of-the-art methods by which encoded Wzx, Wzy, and Wzz proteins can be reliably identified and annotated, using the model Gram-negative bacterium Pseudomonas aeruginosa as an example data set. Lastly, we outline future avenues of research, with the aim of stimulating researchers to take the next steps in investigating the function of, and interplay between, the constituents of this widespread assembly scheme.
Membrane Proteins, lipopolysaccharide (LPS), Wzx flippase, Wzy polymerase, Wzz polysaccharide copolymerase
NCBI PubMed ID: 25358682Publication DOI: 10.1139/cjm-2014-0595Journal NLM ID: 0372707Publisher: National Research Council of Canada
Correspondence: sislam@imm.cnrs.fr; jlam@uoguelph.ca
Institutions: Department of Molecular and Cellular Biology, University of Guelph, Guelph, ON N1G 2W1, Canada
Methods: SDS-PAGE, genetic methods, cloning
- Article ID: 4932
Taylor VL, Hoage JF, Thrane SW, Huszczynski SM, Jelsbak L, Lam JS "A Bacteriophage-Acquired O-Antigen Polymerase (Wzyb) from P. aeruginosa Serotype O16 Performs a Varied Mechanism Compared to Its Cognate Wzya" -
Frontiers in Microbiology 7 (2016) 393
Pseudomonas aeruginosa is a Gram-negative bacterium that produces highly varied lipopolysaccharide (LPS) structures. The O antigen (O-Ag) in the LPS is synthesized through the Wzx/Wzy-dependent pathway where lipid-linked O-Ag repeats are polymerized by Wzy. Horizontal-gene transfer has been associated with O-Ag diversity. The O-Ag present on the surface of serotypes O5 and O16, differ in the intra-molecular bonds, alpha and beta, respectively; the latter arose from the action of three genes in a serotype converting unit acquired from bacteriophage D3, including a beta-polymerase (Wzyβ). To further our understanding of O-polymerases, the inner membrane (IM) topology of Wzyβ was determined using a dual phoA-lacZα reporter system wherein random 3' gene truncations were localized to specific loci with respect to the IM by normalized reporter activities as determined through the ratio of alkaline phosphatase activity to β-galactosidase activity. The topology of Wzyβ developed through this approach was shown to contain two predominant periplasmic loops, PL3 (containing an RX10G motif) and PL4 (having an O-Ag ligase superfamily motif), associated with inverting glycosyltransferase reaction. Through site-directed mutagenesis and complementation assays, residues Arg(254), Arg(270), Arg(272), and His(300) were found to be essential for Wzyβ function. Additionally, like-charge substitutions, R254K and R270K, could not complement the wzyβ knockout, highlighting the essential guanidium side group of Arg residues. The O-Ag ligase domain is conserved among heterologous Wzy proteins that produce β-linked O-Ag repeat units. Taking advantage of the recently obtained whole-genome sequence of serotype O16 a candidate promoter was identified. Wzyβ under its native promoter was integrated in the PAO1 genome, which resulted in simultaneous production of α- and β-linked O-Ag. These observations established that members of Wzy-like family consistently exhibit a dual-periplasmic loops topology, and identifies motifs that are plausible to be involved in enzymatic activities. Based on these results, the phage-derived Wzyβ utilizes a different reaction mechanism in the P. aeruginosa host to avoid self-inhibition during serotype conversion.
Lipopolysaccharide, serotype, Pseudomonas aeruginosa, glycosyltransferase, bacteriophage, O-antigen biosynthesis, polymerase
NCBI PubMed ID: 27065964Publication DOI: 10.3389/fmicb.2016.00393Journal NLM ID: 101548977Publisher: Lausanne: Frontiers Research Foundation
Correspondence: jlam@uoguelph.ca
Institutions: Department of Molecular and Cellular Biology, University of Guelph Guelph, ON, Canada, Department of Systems Biology, Technical University of Denmark Kongens Lyngby, Denmark
Methods: PCR, SDS-PAGE, Western blotting, genetic methods, enzymatic assay
- Article ID: 6301
Qin CJ, Ding MR, Tian GZ, Zou XP, Fu JJ, Hu J, Yin J "Chemical approaches towards installation of rare functional groups in bacterial surface glycans" -
Chinese Journal of Natural Medicines = Zhongguo Tianran Yaowu 20(6) (2022) 401-420
Bacterial surface glycans perform a diverse and important set of biological roles, and have been widely used in the treatment of bacterial infectious diseases. The majority of bacterial surface glycans are decorated with diverse rare functional groups, including amido, acetamidino, carboxamido and pyruvate groups. These functional groups are thought to be important constituents for the biological activities of glycans. Chemical synthesis of glycans bearing these functional groups or their variants is essential for the investigation of structure-activity relationships by a medicinal chemistry approach. To date, a broad choice of synthetic methods is available for targeting the different rare functional groups in bacterial surface glycans. This article reviews the structures of naturally occurring rare functional groups in bacterial surface glycans, and the chemical methods used for installation of these groups.
chemical synthesis, acetamidino group, amido group, bacterial surface glycan, carboxamido group, pyruvyl ketal
NCBI PubMed ID: 35750381Publication DOI: 10.1016/S1875-5364(22)60177-8Journal NLM ID: 101504416Publisher: Beijing: Science Press; Elsevier
Correspondence: J. Yin
Institutions: Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, China, Wuxi School of Medicine, Jiangnan University, Wuxi, China
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5. Compound ID: 962
|
-4)-b-D-ManpNAc3NAmA-(1-4)-b-D-ManpNAc3NAcA-(1-3)-b-D-FucpNAc4Ac-(1- |
Show graphically |
Structure type: polymer chemical repeating unit
Compound class: O-polysaccharide, O-antigen
The structure is contained in the following publication(s):
- Article ID: 254
Hatano K, Pier GB "Complex serology and immune response of mice to variant high-molecular-weight O polysaccharide isolated from Pseudomonas aeruginosa serogroup O2 strains" -
Infection and Immunity 66(8) (1998) 3719-3726
The O antigen of the Pseudomonas aeruginosa lipopolysaccharide is the optimal target for protective antibodies, but the unusual and complex nature of their sugar substituents has made it difficult to define the range of these structures needed in an effective vaccine. Most clinical isolates of P. aeruginosa can be classified into 10 O-antigen serogroups, but slight chemical differences among O polysaccharides within a serogroup give rise to subtype epitopes. These epitopes could impact the reactivity of O-antigen-specific antibodies, as well as the susceptibility of a target strain to protective, opsonic antibodies. To define parameters of serogroup and subtype-epitope immunogenicity, antigenicity, and surface expression on P. aeruginosa cells, we prepared high-molecular-weight O-polysaccharide vaccines from strains of P. aeruginosa serogroup O2, for which eight structurally variant O antigens expressing six defined subtype epitopes (O2a to O2f) have been identified. A complex pattern of immune responses to these antigens was observed following vaccination of mice. The high-molecular-weight O polysaccharides were generally more immunogenic at low doses (1 and 10 microg) than at a high dose (50 microg) and usually elicited antibodies that opsonized the homologous strain for phagocytic killing. Some of the individual polysaccharides elicited cross-opsonic antibodies to a variable number of strains that express all of the defined serogroup O2 subtype epitopes. Combination into one vaccine of two antigens that individually elicited cross-reactive opsonic antibodies to most members of the O2 serogroup inhibited, instead of enhanced, the production of antibodies broadly reactive with most serogroup O2 subtype strains. Thus, immune responses to P. aeruginosa O antigens may be restricted to a limited range of epitopes on structurally complex O antigens, and combining multiple related antigens into a single vaccine formulation may inhibit the production of those antibodies best able to protect against most P. aeruginosa strains within a given O-antigen serogroup.
Lipopolysaccharide, Pseudomonas, Pseudomonas aeruginosa, O-polysaccharide, serology, serogroup, immune response
NCBI PubMed ID: 9673254Journal NLM ID: 0246127Publisher: American Society for Microbiology
Correspondence: gpier@channing.harvard.edu
Institutions: Charming Laboratory, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115-5804., Charming Laboratory, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School,Boston, Massachusetts 02115-5804.
- Article ID: 337
Newton GJ, Daniels C, Burrows LL, Kropinski AM, Clarke AJ, Lam JS "Three-component-mediated serotype conversion in Pseudomonas aeruginosa by bacteriophage D3" -
Molecular Microbiology 39(5) (2001) 1237-1247
Bacteriophage D3 is capable of lysogenizing Pseudomonas aeruginosa PAO1 (serotype O5), converting the O-antigen from O5 to O16 and O-acetylating the N-acetylfucosamine moiety. To investigate the mechanism of lysogenic conversion, a 3.6 kb fragment from the D3 genome was isolated capable of mediating serotypic conversion identical to the D3 lysogen strain (AK1380). The PAO1 transformants containing this 3.6 kb of D3 DNA exhibited identical lipopolysaccharide (LPS) banding patterns to serotype O16 in silver-stained SDS-PAGE gels and displayed reactivity to an antibody specific for O-acetyl groups. Further analysis led to the identification of three open reading frames (ORFs) required for serotype conversion: an α-polymerase inhibitor (iap); an O-acetylase (oac); and a β-polymerase (wzyβ). The α-polymerase inhibitor (Iap) is capable of inhibiting the assembly of the serotype-specific O5 B-band LPS and allows the phage-encoded β-polymerase (Wzyβ) to form new β-linked B-band LPS. The D3 phage also alters the LPS by the addition of O-acetyl groups to the FucNAc residue in the O-antigen repeat unit by the action of the D3 O-acetylase (Oac). These three components form a simple yet elegant system by which bacteriophage D3 is capable of altering the surface of P. aeruginosa PAO1
serotype, Pseudomonas, Pseudomonas aeruginosa, bacteriophage, conversion, serotype conversion
NCBI PubMed ID: 11251840Journal NLM ID: 8712028Publisher: Blackwell Publishing
Correspondence: jlam@uoguelph.ca
Institutions: Department of Microbiology, University of Guelph, Guelph, Ontario, N1G 2W1, Canada, The Centre for Infection and Biomaterials Research, Toronto General Hospital, Toronto, Ontario, M5G 2C4, Canada, Department of Microbiology and Immunology, Queen's University, Kingston, Ontario, K7L 3N6, Canada
Methods: PCR, SDS-PAGE, DNA techniques, Western blotting, quantification of O-acetylation
- Article ID: 1778
Knirel YA, Kochetkov NK "The structure of lipopolysaccharides of gram-negative bacteria. III. The structure of O-antigens: A review" -
Biochemistry (Moscow) 59(12) (1994) 1325-1383
This review summarizes data on the composition and structure of the O-antigens, the polysaccharide chains of the outer-membrane lipopolysaccharides (LPS) of Gram-negative bacteria defining the immunospecificity of these microbial cells. Special reference is given to some structural features of the O-antigens, such as the presence of unique monosaccharides and noncarbohydrate components, masked regularity, and the occurrence in one microorganism of LPS with structurally different polysaccharide chains. Antigenic relationships between microorganisms belonging to different taxonomic groups are discussed.
structure, O-antigen, chemical composition, bacterial lipopolysaccharides, Salmonella livingstone C1
NCBI PubMed ID: 7533007Journal NLM ID: 0376536Publisher: Nauka/Interperiodica
Institutions: Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia
- Article ID: 1836
Knirel YA, Vinogradov EV, Paramonov NA, Shashkov AS, Kochetkov NK, Stanislavsky ES, Mashilova GM "Antigenic polysaccharides of bacteria. 16. Structure of O-specific polysaccharide chain of Pseudomonas aeruginosa O25 (Wokatsch) lipopolysaccharide" -
Bioorganicheskaya Khimia = Bioorganic Chemistry [Russian] 12(9) (1986) 1263-1267
O-Specific polysaccharide built up of trisaccharide repeating units containing 3-acetamidino-2-acetamido-2,3-dideoxy-D-mannuronic acid (ManNAcAmA), 2,3-diacetamido-2,3-dideoxy-D-mannuronic acid (Man(NAc)2A), N-acetyl-D-fucosamine (FucNAc), and O-acetyl group was obtained on mild acid hydrolysis of P. aeruginosa O25 (Wokatsch classification) lipopolysaccharide. Basing on de-O-acetylation of polysaccharide with aqueous triethylamine accompanied by hydrolysis of acetamidino group to acetamido group, as well as on the 1H and 13C NMR data, the following structure of the repeating unit of the polysaccharide was established: (Formula: see text) P. aeruginosa O25 polysaccharide has the same carbohydrate skeleton as that of P. aeruginosa O3a,b (Lányi classification) and differs from the latter only by the presence of the O-acetyl group at position 4 of N-acetylfucosamine.
NCBI PubMed ID: 2430584Journal NLM ID: 7804941Publisher: Moskva: Nauka
Institutions: N.D. Zelinsky Institute of Organic Chemistry, Academy of Sciences of the USSR, Moscow, Russia
Methods: 13C NMR
- Article ID: 2409
Knirel YA, Paramonov NA, Vinogradov EV, Shashkov AS, Dmitriev BA, Kochetkov NK, Kholodkova EV, Stanislavsky ES "Somatic antigens of Pseudomonas aeruginosa. The structure of O-specific polysaccharide chains of lipopolysaccharides of P. aeruginosa O3 (Lányi), O25 (Wokatsch) and Fisher immunotypes 3 and 7" -
European Journal of Biochemistry 167 (1987) 549-561
O-specific polysaccharides, obtained on mild acid degradation of lipopolysacchrides of the serologically related strains Pseudomonas aeruginosa O3 (Lányi classification), O25 (Wokatsch classification) and immunotypes 3 and 7 (Fisher classification), are built up of trisaccharide repeating units involving 2-acetamido-2,6-dideoxy-D-galactose (N-acetyl-D-fucosamine), 2,3-diacetamido-2,3-dideoxy-D-mannuronic acid or 2,3-diacetamido-2,3-dideoxy-L-guluronic acid and 3-acetamidino-2-acetamido-2,3-dideoxy-D-mannuronic acid or 3-acetamidino-2-acetamido-2,3-dideoxy-L-guluronic acid. Lányi O3(a),3d,3f and Wokatsch O25 polysaccharides contain also O-acetyl groups. On the basis of solvolysis with anhydrous hydrogen fluoride, resulting in trisaccharide fragments with N-acetylfucosamine residue at the reducing terminus, chemical modifications of the acetamidino group (alkaline hydrolysis to the acetamido group or reductive deamination to the ethylamino group), as well as analysis by 1H-NMR (including nuclear Overhauser effect experiments) and 13C-NMR spectroscopy, and fast-atom bombardment mass spectrometry, it was concluded that the repeating units of the polysaccharides have the following structures: (Formula: see text) where HexNAcAmA = α-L-GulNAcAmA (approximately 70%) or β-D-ManNacAMA (approximately 30%). Lányi O3(a),3d,3f polysaccharide involves two types of repeating units, which differ from each other only in the configuration at C-5 of the 3-acetamidino-2-acetamido-2,3-dideoxyuronic acid residue. Lányi O3(a),3c,O3a,3d,3e and Fisher immunotypes 3 and 7 polysaccharides contain, together with the major repeating units shown above, a small proportion of units in which the derivative of α-L-guluronic acid is replaced by the corresponding β-D-manno isomer. The data obtained provide the opportunity to substantiate the serological interrelations between these strains of P. aeruginosa by the presence in the O-specific polysaccharides of common monosaccharides or disaccharide fragments. The distinctions between them stem from the presence or absence of the O-acetyl group, a different configuration of the glycosidic linkage of the N-acetylfucosamine residue and/or a different configuration at C-5 of one or both derivatives of diaminouronic acids.
NCBI PubMed ID: 3115777Journal NLM ID: 0107600Publisher: Oxford, UK: Blackwell Science Ltd. on behalf of the Federation of European Biochemical Societies
Institutions: N.D. Zelinsky Institute of Organic Chemistry, Academy of Sciences of the USSR, Moscow, Russia
Methods: 13C NMR, 1H NMR, FAB-MS
- Article ID: 3207
Knirel YA, Bystrova OV, Kocharova NA, Zähringer U, Pier GB "Conserved and variable structural features of the Pseudomonas aeruginosa lipopolysaccharide" -
Journal of Endotoxin Research 12(6) (2006) 324-336
The review is devoted to recent progress in the structural elucidation of the lipopolysaccharide of the bacterium Pseudomonas aeruginosa, including O-antigen biological repeats, core oligosaccharide, and lipid A. Data on biosynthesis, genetics and serology of the lipopolysaccharide isolated from various P. aeruginosa O-serogroups are discussed in relation to the chemical structures.
O-antigen, Pseudomonas aeruginosa, lipid A, core oligosaccharide, lipopolysaccharide structure, serologic activity
NCBI PubMed ID: 17254386Publication DOI: 10.1179/096805106X118906Journal NLM ID: 9433350Publisher: Maney Publishing
Correspondence: knirel@ioc.ac.ru
Institutions: N.D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia, Research Center Borstel, Leibniz-Center for Medicine and Biosciences, Borstel, Germany, Channing Laboratory, Brigham and Women’s Hospital, Harvard Medical School, Boston, Massachusetts, USA
Methods: NMR, chemical methods, MS, serological methods, genetic methods
- Article ID: 3391
Kaluzny K, Abeyrathne PD, Lam JS "Coexistence of Two Distinct Versions of O-Antigen Polymerase, Wzy-Alpha and Wzy-Beta, in Pseudomonas aeruginosa Serogroup O2 and Their Contributions to Cell Surface Diversity" -
Journal of Bacteriology 189(11) (2007) 4141-4152
Assembly of B-band LPS in Pseudomonas aeruginosa follows a Wzy-dependent pathway, requiring the O-antigen polymerase Wzy, and other proteins. The peptide sequences of the wzyα product from strains of serotypes O2, O5, and O16 are identical; but the O-units in O5 are α-glycosidic-linked, while those in O2 and O16 are β-linked. We hypothesized that a derivative of the D3 bacteriophage wzyβ is present in the chromosomes of O2 and O16, and that this gene is responsible for the β-linkage. By a combination of PCR and primer walking, wzyβ of both serotypes have been amplified and cloned. Both are identical, but only share 87.42% sequence identity with their xenolog in D3. A chromosomal knockout mutant of O16 wzyβ was made and it produces rough LPS devoid of B-band O antigen. The cloned wzyβ is capable of complementing the O16 wzyβ mutant, as well as cross-complementing a wzyα knockout mutant. However, in the latter case, the restored O-antigen was β-linked. Using RT-PCR, we showed that wzyα was transcribed in O2 and O16 strains and was functional since both of these genes could complement the wzyα mutant of O5. With the co-existence of wzyα and wzyβ in O2 and O16 and the B-band O polysaccharides in these being β-linked, we hypothesized that iap, an inhibitor of alpha-polymerase gene, must be present in these serotypes. Indeed, through PCR, TOPO cloning and nucleotide sequencing results, we verified the presence of iap in both O2 and O16 serotypes
serotype, Pseudomonas aeruginosa, O-polysaccharide, serogroup, sequencing, O-antigen polymerase, rough LPS
NCBI PubMed ID: 17384183Journal NLM ID: 2985120RPublisher: American Society for Microbiology
Correspondence: jlam@uoguelph.ca
Institutions: University of Guelph, Department of Molecular and Cellular Biology, Guelph, Ontario, Canada N1G 2W1
Methods: serological methods, genetic methods
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6. Compound ID: 963
|
-4)-b-D-ManpNAc3NAmA-(1-4)-a-L-GulpNAc3NAcA-(1-3)-b-D-FucpNAc-(1- |
Show graphically |
Structure type: polymer chemical repeating unit
Compound class: O-polysaccharide, O-antigen, LPS
The structure is contained in the following publication(s):
- Article ID: 254
Hatano K, Pier GB "Complex serology and immune response of mice to variant high-molecular-weight O polysaccharide isolated from Pseudomonas aeruginosa serogroup O2 strains" -
Infection and Immunity 66(8) (1998) 3719-3726
The O antigen of the Pseudomonas aeruginosa lipopolysaccharide is the optimal target for protective antibodies, but the unusual and complex nature of their sugar substituents has made it difficult to define the range of these structures needed in an effective vaccine. Most clinical isolates of P. aeruginosa can be classified into 10 O-antigen serogroups, but slight chemical differences among O polysaccharides within a serogroup give rise to subtype epitopes. These epitopes could impact the reactivity of O-antigen-specific antibodies, as well as the susceptibility of a target strain to protective, opsonic antibodies. To define parameters of serogroup and subtype-epitope immunogenicity, antigenicity, and surface expression on P. aeruginosa cells, we prepared high-molecular-weight O-polysaccharide vaccines from strains of P. aeruginosa serogroup O2, for which eight structurally variant O antigens expressing six defined subtype epitopes (O2a to O2f) have been identified. A complex pattern of immune responses to these antigens was observed following vaccination of mice. The high-molecular-weight O polysaccharides were generally more immunogenic at low doses (1 and 10 microg) than at a high dose (50 microg) and usually elicited antibodies that opsonized the homologous strain for phagocytic killing. Some of the individual polysaccharides elicited cross-opsonic antibodies to a variable number of strains that express all of the defined serogroup O2 subtype epitopes. Combination into one vaccine of two antigens that individually elicited cross-reactive opsonic antibodies to most members of the O2 serogroup inhibited, instead of enhanced, the production of antibodies broadly reactive with most serogroup O2 subtype strains. Thus, immune responses to P. aeruginosa O antigens may be restricted to a limited range of epitopes on structurally complex O antigens, and combining multiple related antigens into a single vaccine formulation may inhibit the production of those antibodies best able to protect against most P. aeruginosa strains within a given O-antigen serogroup.
Lipopolysaccharide, Pseudomonas, Pseudomonas aeruginosa, O-polysaccharide, serology, serogroup, immune response
NCBI PubMed ID: 9673254Journal NLM ID: 0246127Publisher: American Society for Microbiology
Correspondence: gpier@channing.harvard.edu
Institutions: Charming Laboratory, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115-5804., Charming Laboratory, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School,Boston, Massachusetts 02115-5804.
- Article ID: 337
Newton GJ, Daniels C, Burrows LL, Kropinski AM, Clarke AJ, Lam JS "Three-component-mediated serotype conversion in Pseudomonas aeruginosa by bacteriophage D3" -
Molecular Microbiology 39(5) (2001) 1237-1247
Bacteriophage D3 is capable of lysogenizing Pseudomonas aeruginosa PAO1 (serotype O5), converting the O-antigen from O5 to O16 and O-acetylating the N-acetylfucosamine moiety. To investigate the mechanism of lysogenic conversion, a 3.6 kb fragment from the D3 genome was isolated capable of mediating serotypic conversion identical to the D3 lysogen strain (AK1380). The PAO1 transformants containing this 3.6 kb of D3 DNA exhibited identical lipopolysaccharide (LPS) banding patterns to serotype O16 in silver-stained SDS-PAGE gels and displayed reactivity to an antibody specific for O-acetyl groups. Further analysis led to the identification of three open reading frames (ORFs) required for serotype conversion: an α-polymerase inhibitor (iap); an O-acetylase (oac); and a β-polymerase (wzyβ). The α-polymerase inhibitor (Iap) is capable of inhibiting the assembly of the serotype-specific O5 B-band LPS and allows the phage-encoded β-polymerase (Wzyβ) to form new β-linked B-band LPS. The D3 phage also alters the LPS by the addition of O-acetyl groups to the FucNAc residue in the O-antigen repeat unit by the action of the D3 O-acetylase (Oac). These three components form a simple yet elegant system by which bacteriophage D3 is capable of altering the surface of P. aeruginosa PAO1
serotype, Pseudomonas, Pseudomonas aeruginosa, bacteriophage, conversion, serotype conversion
NCBI PubMed ID: 11251840Journal NLM ID: 8712028Publisher: Blackwell Publishing
Correspondence: jlam@uoguelph.ca
Institutions: Department of Microbiology, University of Guelph, Guelph, Ontario, N1G 2W1, Canada, The Centre for Infection and Biomaterials Research, Toronto General Hospital, Toronto, Ontario, M5G 2C4, Canada, Department of Microbiology and Immunology, Queen's University, Kingston, Ontario, K7L 3N6, Canada
Methods: PCR, SDS-PAGE, DNA techniques, Western blotting, quantification of O-acetylation
- Article ID: 1778
Knirel YA, Kochetkov NK "The structure of lipopolysaccharides of gram-negative bacteria. III. The structure of O-antigens: A review" -
Biochemistry (Moscow) 59(12) (1994) 1325-1383
This review summarizes data on the composition and structure of the O-antigens, the polysaccharide chains of the outer-membrane lipopolysaccharides (LPS) of Gram-negative bacteria defining the immunospecificity of these microbial cells. Special reference is given to some structural features of the O-antigens, such as the presence of unique monosaccharides and noncarbohydrate components, masked regularity, and the occurrence in one microorganism of LPS with structurally different polysaccharide chains. Antigenic relationships between microorganisms belonging to different taxonomic groups are discussed.
structure, O-antigen, chemical composition, bacterial lipopolysaccharides, Salmonella livingstone C1
NCBI PubMed ID: 7533007Journal NLM ID: 0376536Publisher: Nauka/Interperiodica
Institutions: Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia
- Article ID: 1833
Knirel YA, Paramonov NA, Vinogradov EV, Shashkov AS, Dmitriev BA, Kochetkov NK "Structure of Pseudomonas aeruginosa immunotype 3 O-specific polysaccharide: revision of the structure of acetamidino derivative of 2,3-diamino-2,3-dideoxy-D-mannuronic acid" -
Bioorganicheskaya Khimia = Bioorganic Chemistry [Russian] 12(7) (1986) 995-997
O-Specific side chain of P. aeruginosa immunotype 3 lipopolysaccharide is composed of N-acetyl-D-fucosamine (FucNAc), 2,3-diacetamido-2,3-dideoxy-L-guluronic acid (GulN2Ac2A) and 3-acetamidino = 2-acetamido = 2,3 = dideoxy = D-mannuronic acid (ManNAcAmA). The latter sugar is identified on the basis of solvolysis with anhydrous hydrogen fluoride, 13C NMR spectroscopy and fast-atom bombardment mass spectrometry analysis, as well as of reactions of acetamidino function (alkaline hydrolysis to acetamido group and reductive deamination to ethylamino group). Earlier, in the course of investigation of P. aeruginosa O3 lipopolysaccharides, the structure of 1-methyl-2-imidazoline was erroneously ascribed to the acetamidino group. The following structure was established for the repeating unit of immunotype 3 polysaccharide which is identical to P. aeruginosa O3(a),3c polysaccharide: →4)-β-D-ManNAcAmA-(1→4)-α-L-GulN2Ac2A-(1→3)-β-D-FucNac-(1→.
NCBI PubMed ID: 2429671Journal NLM ID: 7804941Publisher: Moskva: Nauka
Institutions: N.D. Zelinsky Institute of Organic Chemistry, Academy of Sciences of the USSR, Moscow, Russia
Methods: 13C NMR
- Article ID: 1843
Elkin YN, Knirel YA, Vinogradov EV, Paramonov NA, Troshkov ML, Aminev "Fast-atom-bombardment mass-spectra of aminooligosaccharides" -
Bioorganicheskaya Khimia = Bioorganic Chemistry [Russian] 12 (1986) 1658-1661
Journal NLM ID: 7804941Publisher: Moskva: Nauka
Institutions: N.D. Zelinsky Institute of Organic Chemistry, Academy of Sciences of the USSR, Moscow, Russia
Methods: FAB-MS
- Article ID: 1844
Knirel YA, Kocharova NA, Vinogradov EV, Paramonov NA, Dmitriev BA, Kochetkov NK, Stanislavsky ES, Kholodkova EV "Antigenic polysaccharides of bacteria. 21. Structure of O-specific polysaccharide chains and serological specificity of lipopolysaccharides of seven Pseudomonas aeruginosa immunotypes" -
Bioorganicheskaya Khimia = Bioorganic Chemistry [Russian] 13 (1987) 88-96
On mild acid degradation on lipopolysaccharides of seven Pseudomonas aeruginosa immunotypes, O-specific polysaccharides were obtained and their structures established. A peculiar feature of the polysaccharides is the presence of various, mostly acidic, mono- and diaminosugars, many of which have not previously been found in nature. The absence of serological cross-reactions (inhibition of passive haemagglutination) between lipopolysaccharides of seven immunotypes correlates with the absence of any common oligosaccharide fragments in their O-specific chains. The data obtained revealed structural and serological interrelations between O-antigens of seven immunotypes and P. aeruginosa O-serotypes, and showed that immunotypes 1 and 7 should be included into the serological classification scheme as individual O-serotypes.
NCBI PubMed ID: 2436629Journal NLM ID: 7804941Publisher: Moskva: Nauka
Institutions: N.D. Zelinsky Institute of Organic Chemistry, Academy of Sciences of the USSR, Moscow, Russia
- Article ID: 2409
Knirel YA, Paramonov NA, Vinogradov EV, Shashkov AS, Dmitriev BA, Kochetkov NK, Kholodkova EV, Stanislavsky ES "Somatic antigens of Pseudomonas aeruginosa. The structure of O-specific polysaccharide chains of lipopolysaccharides of P. aeruginosa O3 (Lányi), O25 (Wokatsch) and Fisher immunotypes 3 and 7" -
European Journal of Biochemistry 167 (1987) 549-561
O-specific polysaccharides, obtained on mild acid degradation of lipopolysacchrides of the serologically related strains Pseudomonas aeruginosa O3 (Lányi classification), O25 (Wokatsch classification) and immunotypes 3 and 7 (Fisher classification), are built up of trisaccharide repeating units involving 2-acetamido-2,6-dideoxy-D-galactose (N-acetyl-D-fucosamine), 2,3-diacetamido-2,3-dideoxy-D-mannuronic acid or 2,3-diacetamido-2,3-dideoxy-L-guluronic acid and 3-acetamidino-2-acetamido-2,3-dideoxy-D-mannuronic acid or 3-acetamidino-2-acetamido-2,3-dideoxy-L-guluronic acid. Lányi O3(a),3d,3f and Wokatsch O25 polysaccharides contain also O-acetyl groups. On the basis of solvolysis with anhydrous hydrogen fluoride, resulting in trisaccharide fragments with N-acetylfucosamine residue at the reducing terminus, chemical modifications of the acetamidino group (alkaline hydrolysis to the acetamido group or reductive deamination to the ethylamino group), as well as analysis by 1H-NMR (including nuclear Overhauser effect experiments) and 13C-NMR spectroscopy, and fast-atom bombardment mass spectrometry, it was concluded that the repeating units of the polysaccharides have the following structures: (Formula: see text) where HexNAcAmA = α-L-GulNAcAmA (approximately 70%) or β-D-ManNacAMA (approximately 30%). Lányi O3(a),3d,3f polysaccharide involves two types of repeating units, which differ from each other only in the configuration at C-5 of the 3-acetamidino-2-acetamido-2,3-dideoxyuronic acid residue. Lányi O3(a),3c,O3a,3d,3e and Fisher immunotypes 3 and 7 polysaccharides contain, together with the major repeating units shown above, a small proportion of units in which the derivative of α-L-guluronic acid is replaced by the corresponding β-D-manno isomer. The data obtained provide the opportunity to substantiate the serological interrelations between these strains of P. aeruginosa by the presence in the O-specific polysaccharides of common monosaccharides or disaccharide fragments. The distinctions between them stem from the presence or absence of the O-acetyl group, a different configuration of the glycosidic linkage of the N-acetylfucosamine residue and/or a different configuration at C-5 of one or both derivatives of diaminouronic acids.
NCBI PubMed ID: 3115777Journal NLM ID: 0107600Publisher: Oxford, UK: Blackwell Science Ltd. on behalf of the Federation of European Biochemical Societies
Institutions: N.D. Zelinsky Institute of Organic Chemistry, Academy of Sciences of the USSR, Moscow, Russia
Methods: 13C NMR, 1H NMR, FAB-MS
- Article ID: 3207
Knirel YA, Bystrova OV, Kocharova NA, Zähringer U, Pier GB "Conserved and variable structural features of the Pseudomonas aeruginosa lipopolysaccharide" -
Journal of Endotoxin Research 12(6) (2006) 324-336
The review is devoted to recent progress in the structural elucidation of the lipopolysaccharide of the bacterium Pseudomonas aeruginosa, including O-antigen biological repeats, core oligosaccharide, and lipid A. Data on biosynthesis, genetics and serology of the lipopolysaccharide isolated from various P. aeruginosa O-serogroups are discussed in relation to the chemical structures.
O-antigen, Pseudomonas aeruginosa, lipid A, core oligosaccharide, lipopolysaccharide structure, serologic activity
NCBI PubMed ID: 17254386Publication DOI: 10.1179/096805106X118906Journal NLM ID: 9433350Publisher: Maney Publishing
Correspondence: knirel@ioc.ac.ru
Institutions: N.D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia, Research Center Borstel, Leibniz-Center for Medicine and Biosciences, Borstel, Germany, Channing Laboratory, Brigham and Women’s Hospital, Harvard Medical School, Boston, Massachusetts, USA
Methods: NMR, chemical methods, MS, serological methods, genetic methods
- Article ID: 3391
Kaluzny K, Abeyrathne PD, Lam JS "Coexistence of Two Distinct Versions of O-Antigen Polymerase, Wzy-Alpha and Wzy-Beta, in Pseudomonas aeruginosa Serogroup O2 and Their Contributions to Cell Surface Diversity" -
Journal of Bacteriology 189(11) (2007) 4141-4152
Assembly of B-band LPS in Pseudomonas aeruginosa follows a Wzy-dependent pathway, requiring the O-antigen polymerase Wzy, and other proteins. The peptide sequences of the wzyα product from strains of serotypes O2, O5, and O16 are identical; but the O-units in O5 are α-glycosidic-linked, while those in O2 and O16 are β-linked. We hypothesized that a derivative of the D3 bacteriophage wzyβ is present in the chromosomes of O2 and O16, and that this gene is responsible for the β-linkage. By a combination of PCR and primer walking, wzyβ of both serotypes have been amplified and cloned. Both are identical, but only share 87.42% sequence identity with their xenolog in D3. A chromosomal knockout mutant of O16 wzyβ was made and it produces rough LPS devoid of B-band O antigen. The cloned wzyβ is capable of complementing the O16 wzyβ mutant, as well as cross-complementing a wzyα knockout mutant. However, in the latter case, the restored O-antigen was β-linked. Using RT-PCR, we showed that wzyα was transcribed in O2 and O16 strains and was functional since both of these genes could complement the wzyα mutant of O5. With the co-existence of wzyα and wzyβ in O2 and O16 and the B-band O polysaccharides in these being β-linked, we hypothesized that iap, an inhibitor of alpha-polymerase gene, must be present in these serotypes. Indeed, through PCR, TOPO cloning and nucleotide sequencing results, we verified the presence of iap in both O2 and O16 serotypes
serotype, Pseudomonas aeruginosa, O-polysaccharide, serogroup, sequencing, O-antigen polymerase, rough LPS
NCBI PubMed ID: 17384183Journal NLM ID: 2985120RPublisher: American Society for Microbiology
Correspondence: jlam@uoguelph.ca
Institutions: University of Guelph, Department of Molecular and Cellular Biology, Guelph, Ontario, Canada N1G 2W1
Methods: serological methods, genetic methods
- Article ID: 3711
King JD, Kocincova D, Westman EL, Lam JS "Lipopolysaccharide biosynthesis in Pseudomonas aeruginosa" -
Innate Immunity 15(5) (2009) 261-312
Pseudomonas aeruginosa causes serious nosocomial infections, and an important virulence factor produced by this organism is lipopolysaccharide (LPS). This review summarizes knowledge about biosynthesis of all three structural domains of LPS - lipid A, core oligosaccharide, and O polysaccharides. In addition, based on similarities with other bacterial species, this review proposes new hypothetical pathways for unstudied steps in the biosynthesis of P. aeruginosa LPS. Lipid A biosynthesis is discussed in relation to Escherichia coli and Salmonella, and the biosyntheses of core sugar precursors and core oligosaccharide are summarised. Pseudomonas aeruginosa attaches a Common Polysaccharide Antigen and O-Specific Antigen polysaccharides to lipid A-core. Both forms of O polysaccharide are discussed with respect to their independent synthesis mechanisms. Recent advances in understanding O-polysaccharide biosynthesis since the last major review on this subject, published nearly a decade ago, are highlighted. Since P. aeruginosa O polysaccharides contain unusual sugars, sugar-nucleotide biosynthesis pathways are reviewed in detail. Knowledge derived from detailed studies in the O5, O6 and O11 serotypes is applied to predict biosynthesis pathways of sugars in poorly-studied serotypes, especially O1, O4, and O13/O14. Although further work is required, a full understanding of LPS biosynthesis in P. aeruginosa is almost within reach.
Lipopolysaccharide, core, O-antigen, Pseudomonas aeruginosa, lipid A
NCBI PubMed ID: 19710102Publication DOI: 10.1177/1753425909106436Journal NLM ID: 101469670Publisher: Sage Publications
Correspondence: jlam@uoguelph.ca
Institutions: Department of Molecular and Cellular Biology, University of Guelph, Guelph, Ontario, Canada
- Article ID: 4062
Lam JS, Taylor VL, Islam ST, Hao Y, Kocincova D "Genetic and Functional Diversity of Pseudomonas aeruginosa Lipopolysaccharide" -
Frontiers in Microbiology 2 (2011) 118
Lipopolysccharide (LPS) is an integral component of the Pseudomonas aeruginosa cell envelope, occupying the outer leaflet of the outer membrane in this Gram-negative opportunistic pathogen. It is important for bacterium-host interactions and has been shown to be a major virulence factor for this organism. Structurally, P. aeruginosa LPS is composed of three domains, namely, lipid A, core oligosaccharide, and the distal O antigen (O-Ag). Most P. aeruginosa strains produce two distinct forms of O-Ag, one a homopolymer of D-rhamnose that is a common polysaccharide antigen (CPA, formerly termed A band), and the other a heteropolymer of three to five distinct (and often unique dideoxy) sugars in its repeat units, known as O-specific antigen (OSA, formerly termed B band). Compositional differences in the O units among the OSA from different strains form the basis of the International Antigenic Typing Scheme for classification via serotyping of different strains of P. aeruginosa. The focus of this review is to provide state-of-the-art knowledge on the genetic and resultant functional diversity of LPS produced by P. aeruginosa. The underlying factors contributing to this diversity will be thoroughly discussed and presented in the context of its contributions to host-pathogen interactions and the control/prevention of infection.
Lipopolysaccharide, biosynthesis, virulence, serotyping, bacteriophage, motility, seroconversion, nucleotide sugars
NCBI PubMed ID: 21687428Publication DOI: 10.3389/fmicb.2011.00118Journal NLM ID: 101548977Publisher: Lausanne: Frontiers Research Foundation
Correspondence: jlam@uoguelph.ca
Institutions: Department of Molecular and Cellular Biology, University of Guelph Guelph, ON, Canada
- Article ID: 4684
Islam ST, Lam JS "Synthesis of bacterial polysaccharides via the Wzx/Wzy-dependent pathway" -
Canadian Journal of Microbiology 60(11) (2014) 697-716
The surfaces of bacteria mediate a multitude of functions in the environment and in an infected host, including adhesion to both biotic and abiotic substrata, motility, immune system interaction and (or) activation, biofilm formation, and cell-cell communication, with many of these features directly influenced by cell-surface glycans. In both Gram-negative and Gram-positive bacteria, the majority of cell-surface polysaccharides are produced via the Wzx/Wzy-dependent assembly pathway; these glycans include heteropolymeric O-antigen, enterobacterial common antigen, exopolysaccharide, spore coat, and capsule in diverse bacteria. The key components of this assembly pathway are the integral inner membrane Wzx flippase, Wzy polymerase, and Wzz chain-length regulator proteins, which until recently have resisted detailed structural and functional characterization. In this review, we have provided a comprehensive synthesis of the latest structural and mechanistic data for each protein, as well as an examination of substrate specificity for each assembly step and complex formation between the constituent proteins. To complement the unprecedented explosion of genomic-sequencing data for bacteria, we have also highlighted both classical and state-of-the-art methods by which encoded Wzx, Wzy, and Wzz proteins can be reliably identified and annotated, using the model Gram-negative bacterium Pseudomonas aeruginosa as an example data set. Lastly, we outline future avenues of research, with the aim of stimulating researchers to take the next steps in investigating the function of, and interplay between, the constituents of this widespread assembly scheme.
Membrane Proteins, lipopolysaccharide (LPS), Wzx flippase, Wzy polymerase, Wzz polysaccharide copolymerase
NCBI PubMed ID: 25358682Publication DOI: 10.1139/cjm-2014-0595Journal NLM ID: 0372707Publisher: National Research Council of Canada
Correspondence: sislam@imm.cnrs.fr; jlam@uoguelph.ca
Institutions: Department of Molecular and Cellular Biology, University of Guelph, Guelph, ON N1G 2W1, Canada
Methods: SDS-PAGE, genetic methods, cloning
- Article ID: 4932
Taylor VL, Hoage JF, Thrane SW, Huszczynski SM, Jelsbak L, Lam JS "A Bacteriophage-Acquired O-Antigen Polymerase (Wzyb) from P. aeruginosa Serotype O16 Performs a Varied Mechanism Compared to Its Cognate Wzya" -
Frontiers in Microbiology 7 (2016) 393
Pseudomonas aeruginosa is a Gram-negative bacterium that produces highly varied lipopolysaccharide (LPS) structures. The O antigen (O-Ag) in the LPS is synthesized through the Wzx/Wzy-dependent pathway where lipid-linked O-Ag repeats are polymerized by Wzy. Horizontal-gene transfer has been associated with O-Ag diversity. The O-Ag present on the surface of serotypes O5 and O16, differ in the intra-molecular bonds, alpha and beta, respectively; the latter arose from the action of three genes in a serotype converting unit acquired from bacteriophage D3, including a beta-polymerase (Wzyβ). To further our understanding of O-polymerases, the inner membrane (IM) topology of Wzyβ was determined using a dual phoA-lacZα reporter system wherein random 3' gene truncations were localized to specific loci with respect to the IM by normalized reporter activities as determined through the ratio of alkaline phosphatase activity to β-galactosidase activity. The topology of Wzyβ developed through this approach was shown to contain two predominant periplasmic loops, PL3 (containing an RX10G motif) and PL4 (having an O-Ag ligase superfamily motif), associated with inverting glycosyltransferase reaction. Through site-directed mutagenesis and complementation assays, residues Arg(254), Arg(270), Arg(272), and His(300) were found to be essential for Wzyβ function. Additionally, like-charge substitutions, R254K and R270K, could not complement the wzyβ knockout, highlighting the essential guanidium side group of Arg residues. The O-Ag ligase domain is conserved among heterologous Wzy proteins that produce β-linked O-Ag repeat units. Taking advantage of the recently obtained whole-genome sequence of serotype O16 a candidate promoter was identified. Wzyβ under its native promoter was integrated in the PAO1 genome, which resulted in simultaneous production of α- and β-linked O-Ag. These observations established that members of Wzy-like family consistently exhibit a dual-periplasmic loops topology, and identifies motifs that are plausible to be involved in enzymatic activities. Based on these results, the phage-derived Wzyβ utilizes a different reaction mechanism in the P. aeruginosa host to avoid self-inhibition during serotype conversion.
Lipopolysaccharide, serotype, Pseudomonas aeruginosa, glycosyltransferase, bacteriophage, O-antigen biosynthesis, polymerase
NCBI PubMed ID: 27065964Publication DOI: 10.3389/fmicb.2016.00393Journal NLM ID: 101548977Publisher: Lausanne: Frontiers Research Foundation
Correspondence: jlam@uoguelph.ca
Institutions: Department of Molecular and Cellular Biology, University of Guelph Guelph, ON, Canada, Department of Systems Biology, Technical University of Denmark Kongens Lyngby, Denmark
Methods: PCR, SDS-PAGE, Western blotting, genetic methods, enzymatic assay
- Article ID: 5760
Dobrochaeva K, Khasbiulina N, Shilova N, Antipova N, Obukhova P, Galanina O, Blixt O, Kunz H, Filatov A, Knirel Y, Le Pendu J, Khaidukov S, Bovin N "Specificity of human natural antibodies referred to as anti-Tn" -
Molecular Immunology 120 (2020) 74-82
To understand the role of human natural IgM known as antibodies against the carbohydrate epitope Tn, the antibodies were isolated using GalNAcα-Sepharose affinity chromatography, and their specificity was profiled using microarrays (a glycan array printed with oligosaccharides and bacterial polysaccharides, as well as a glycopeptide array), flow cytometry, and inhibition ELISA. The antibodies bound a restricted number of GalNAcα-terminated oligosaccharides better than the parent monosaccharide, e.g., 6-O-Su-GalNAcα and GalNAcα1-3Galβ1-3(4)GlcNAcβ. The binding with several bacterial polysaccharides that have no structural resemblance to the affinity ligand GalNAcα was quite unexpected. Given that GalNAcα is considered the key fragment of the Tn antigen, it is surprising that these antibodies bind weakly GalNAcα-OSer and do not bind a wide variety of GalNAcα-OSer/Thr-containing mucin glycopeptides. At the same time, we have observed specific binding to cells having Tn-positive glycoproteins containing similar glycopeptide motifs in a conformationally rigid macromolecule. Thus, specific recognition of the Tn antigen apparently requires that the naturally occurring "anti-Tn" IgM recognize a complex epitope comprising the GalNAcα as an essential component and a fairly long amino acid sequence where the amino acids adjacent to GalNAcα do not contact the antibody paratope; i.e., the antibodies recognize a spatial epitope or a molecular pattern rather than a classical continuous sequence. In addition, we have not found any increase in the binding of natural antibodies when GalNAcα residues were clustered. These results may help in further development of anticancer vaccines based on synthetic Tn constructs.
cancer, glycans, natural antibodies, anti-glycan antibodies, Tn antigen
NCBI PubMed ID: 32087569Publication DOI: 10.1016/j.molimm.2020.02.005Journal NLM ID: 7905289Publisher: Elsevier
Correspondence: professorbovin@yandex.ru
Institutions: Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 16/10 Miklukho-Maklaya, Moscow, Russian Federation, Semiotik LLC, 16/10 Miklukho-Maklaya, Moscow, Russian Federation, National Medical Research Center for Obstetrics, Gynecology and Perinatology Named after Academician V.I. Kulakov of the Ministry of Healthcare of Russian Federation, Moscow, Russian Federation, National Research University Higher School of Economics, Moscow, Russian Federation, Department of Chemistry, Chemical Biology, University of Copenhagen, Thorvaldsensvej 40, 1871 Frederiksberg C, Denmark, Institut Fur Organische Chemie, Johannes Gutenberg-Universitat Mainz, Duesbergweg 10-14, D-55128, Mainz, Germany, Institute of Immunology, Federal Medical-Biological Agency of Russia, Moscow, Russian Federation, University of Nantes, Inserm, U892 IRT UN, 8 Quai MonCousu, BP70721 Nantes, FR 44007, France
Methods: ELISA, affinity chromatography, flow cytometry analysis, printed glycan array (PGA) analysis, FACS assay
- Article ID: 6301
Qin CJ, Ding MR, Tian GZ, Zou XP, Fu JJ, Hu J, Yin J "Chemical approaches towards installation of rare functional groups in bacterial surface glycans" -
Chinese Journal of Natural Medicines = Zhongguo Tianran Yaowu 20(6) (2022) 401-420
Bacterial surface glycans perform a diverse and important set of biological roles, and have been widely used in the treatment of bacterial infectious diseases. The majority of bacterial surface glycans are decorated with diverse rare functional groups, including amido, acetamidino, carboxamido and pyruvate groups. These functional groups are thought to be important constituents for the biological activities of glycans. Chemical synthesis of glycans bearing these functional groups or their variants is essential for the investigation of structure-activity relationships by a medicinal chemistry approach. To date, a broad choice of synthetic methods is available for targeting the different rare functional groups in bacterial surface glycans. This article reviews the structures of naturally occurring rare functional groups in bacterial surface glycans, and the chemical methods used for installation of these groups.
chemical synthesis, acetamidino group, amido group, bacterial surface glycan, carboxamido group, pyruvyl ketal
NCBI PubMed ID: 35750381Publication DOI: 10.1016/S1875-5364(22)60177-8Journal NLM ID: 101504416Publisher: Beijing: Science Press; Elsevier
Correspondence: J. Yin
Institutions: Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, China, Wuxi School of Medicine, Jiangnan University, Wuxi, China
Expand this compound
Collapse this compound
7. Compound ID: 964
|
-4)-b-D-ManpNAc3NAmA-(1-4)-b-D-ManpNAc3NAcA-(1-3)-a-D-FucpNAc-(1- |
Show graphically |
Structure type: polymer chemical repeating unit
Trivial name: B band polysaccharide, B-band polysaccharide
Compound class: O-polysaccharide, O-antigen
Contained glycoepitopes: IEDB_2275410
The structure is contained in the following publication(s):
- Article ID: 254
Hatano K, Pier GB "Complex serology and immune response of mice to variant high-molecular-weight O polysaccharide isolated from Pseudomonas aeruginosa serogroup O2 strains" -
Infection and Immunity 66(8) (1998) 3719-3726
The O antigen of the Pseudomonas aeruginosa lipopolysaccharide is the optimal target for protective antibodies, but the unusual and complex nature of their sugar substituents has made it difficult to define the range of these structures needed in an effective vaccine. Most clinical isolates of P. aeruginosa can be classified into 10 O-antigen serogroups, but slight chemical differences among O polysaccharides within a serogroup give rise to subtype epitopes. These epitopes could impact the reactivity of O-antigen-specific antibodies, as well as the susceptibility of a target strain to protective, opsonic antibodies. To define parameters of serogroup and subtype-epitope immunogenicity, antigenicity, and surface expression on P. aeruginosa cells, we prepared high-molecular-weight O-polysaccharide vaccines from strains of P. aeruginosa serogroup O2, for which eight structurally variant O antigens expressing six defined subtype epitopes (O2a to O2f) have been identified. A complex pattern of immune responses to these antigens was observed following vaccination of mice. The high-molecular-weight O polysaccharides were generally more immunogenic at low doses (1 and 10 microg) than at a high dose (50 microg) and usually elicited antibodies that opsonized the homologous strain for phagocytic killing. Some of the individual polysaccharides elicited cross-opsonic antibodies to a variable number of strains that express all of the defined serogroup O2 subtype epitopes. Combination into one vaccine of two antigens that individually elicited cross-reactive opsonic antibodies to most members of the O2 serogroup inhibited, instead of enhanced, the production of antibodies broadly reactive with most serogroup O2 subtype strains. Thus, immune responses to P. aeruginosa O antigens may be restricted to a limited range of epitopes on structurally complex O antigens, and combining multiple related antigens into a single vaccine formulation may inhibit the production of those antibodies best able to protect against most P. aeruginosa strains within a given O-antigen serogroup.
Lipopolysaccharide, Pseudomonas, Pseudomonas aeruginosa, O-polysaccharide, serology, serogroup, immune response
NCBI PubMed ID: 9673254Journal NLM ID: 0246127Publisher: American Society for Microbiology
Correspondence: gpier@channing.harvard.edu
Institutions: Charming Laboratory, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115-5804., Charming Laboratory, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School,Boston, Massachusetts 02115-5804.
- Article ID: 337
Newton GJ, Daniels C, Burrows LL, Kropinski AM, Clarke AJ, Lam JS "Three-component-mediated serotype conversion in Pseudomonas aeruginosa by bacteriophage D3" -
Molecular Microbiology 39(5) (2001) 1237-1247
Bacteriophage D3 is capable of lysogenizing Pseudomonas aeruginosa PAO1 (serotype O5), converting the O-antigen from O5 to O16 and O-acetylating the N-acetylfucosamine moiety. To investigate the mechanism of lysogenic conversion, a 3.6 kb fragment from the D3 genome was isolated capable of mediating serotypic conversion identical to the D3 lysogen strain (AK1380). The PAO1 transformants containing this 3.6 kb of D3 DNA exhibited identical lipopolysaccharide (LPS) banding patterns to serotype O16 in silver-stained SDS-PAGE gels and displayed reactivity to an antibody specific for O-acetyl groups. Further analysis led to the identification of three open reading frames (ORFs) required for serotype conversion: an α-polymerase inhibitor (iap); an O-acetylase (oac); and a β-polymerase (wzyβ). The α-polymerase inhibitor (Iap) is capable of inhibiting the assembly of the serotype-specific O5 B-band LPS and allows the phage-encoded β-polymerase (Wzyβ) to form new β-linked B-band LPS. The D3 phage also alters the LPS by the addition of O-acetyl groups to the FucNAc residue in the O-antigen repeat unit by the action of the D3 O-acetylase (Oac). These three components form a simple yet elegant system by which bacteriophage D3 is capable of altering the surface of P. aeruginosa PAO1
serotype, Pseudomonas, Pseudomonas aeruginosa, bacteriophage, conversion, serotype conversion
NCBI PubMed ID: 11251840Journal NLM ID: 8712028Publisher: Blackwell Publishing
Correspondence: jlam@uoguelph.ca
Institutions: Department of Microbiology, University of Guelph, Guelph, Ontario, N1G 2W1, Canada, The Centre for Infection and Biomaterials Research, Toronto General Hospital, Toronto, Ontario, M5G 2C4, Canada, Department of Microbiology and Immunology, Queen's University, Kingston, Ontario, K7L 3N6, Canada
Methods: PCR, SDS-PAGE, DNA techniques, Western blotting, quantification of O-acetylation
- Article ID: 913
Lam JS, Rocchetta HL, Burrows LL "Glycosyltransferases of Pseudomonas aeruginosa that assemble the O antigens of A band and B band lipopolysaccharide" -
Journal of Endotoxin Research (1999) 96-101
Pseudomonas aeruginosa produces two forms of lipopolysaccharide (LPS) designated A band and B band. The O-polysaccharide region of A band is a conserved D-rhamnan polymer arranged alfa1-2,alfa1-3,alfa1-3, while B band is serotype-specific with differences in the O-antigenic region dividing P. aeruginosa into 20 stereotypes. The B band O-antigen unit of serotype O5 is [-4)bDMan(2NAc3N)A(1-4)bDMan(2NAc3NAc)A(1-3)aDFuc2NAc]. The glycosidic structure of LPS molecules specified by the action of dedicated glycosyltransferases. The wbp clusters of A band and B band (serotype O5) were each found to contain three genes coding for putative glycosyltransferases: wbpX, wbpY, wbpZ, and wbpH, wbpJ, wbpL, respectively. To examine the role of these potential transferases in LPS assembly, chromosomal mutaions were generated within all 6 genes. LPS analysis reveals that wbpX, wbpY and wbpZ mutants express and A-B+ phenotype, while wbpH and wbpJ metanst are A+B-. Interstingly, mutations in wbpL, and Escherichia coli wecA homologue, abrogates both A band and B band LPS synthesis, Based on amino acid homologies, O-polysaccharide structures and LPS phenotypes of transferase mutants, we propose and assembly scheme for these two LPS molecules.
Lipopolysaccharide, biosynthesis, genetic, antigen, characterization, O-antigen, B-band, O antigen, Pseudomonas, Pseudomonas aeruginosa, antigens, O antigens, O-antigens, glycosyltransferases, biochemical, review, glycosyltransferase, operon, A-band
Publication DOI: 10.1177/09680519990050011001Journal NLM ID: 9433350Publisher: Maney Publishing
Correspondence: jlam@uoguelph.ca
Institutions: Canadian Bacterial Diseases Network, Department of Microbiology, University of Gueoph, Gueoph, Ontario, Canada
- Article ID: 1395
Burrows LL, Pigeon KE, Lam JS "Psudomonas aeruginosa B-band lipopolysaccharide genes wbpA and wbpI and their Escherichia coli homologues wecC and wecB are not functionally interchangeable" -
FEMS Microbiology Reviews 189 (2000) 135-141
The O antigen unit of Pseudomonas aeruginosa serotype O5 is a complex trisaccharide containing 2-acetamido-3-acetiminido-2,3-dideoxy- L-D-mannuronic acid, 2-acetimido-3-acetimido-2,3-dideoxy-L-D-mannuronic acid, and 2-acetimido-2,6-deoxy-L-D-galactosamine. Specific knockout mutations in the putative UDP-D-N-acetylglucosamine (UDP-D-GlcNAc) epimerase gene, wbpI, or the putative UDP-D-Nacetylmannosamine dehydrogenase gene, wbpA, resulted in strains that no longer produced B-band lipopolysaccharide, confirming the essential roles of these genes in B-band O antigen synthesis. Despite approximately 50% similarity of wbpI and wbpA to the Escherichia coli genes wecB (rffE) and wecC (rffD) involved in enterobacterial common antigen synthesis, cross-complementation experiments were not successful. These results imply that the P. aeruginosa UDP-D-GlcNAc precursor may be di-N-acetylated prior to further modification, preventing the E. coli enzymes from recognizing it as a substrate.
Lipopolysaccharide, gene, B-band, Escherichia, Escherichia coli, WbpA, WbpI
NCBI PubMed ID: 10930727Journal NLM ID: 8902526Publisher: Oxford University Press
Correspondence: jlam@uoguelph.ca
Institutions: Department of Microbiology, University of Guelph, Gueph, Canada
Methods: genetic methods
- Article ID: 2409
Knirel YA, Paramonov NA, Vinogradov EV, Shashkov AS, Dmitriev BA, Kochetkov NK, Kholodkova EV, Stanislavsky ES "Somatic antigens of Pseudomonas aeruginosa. The structure of O-specific polysaccharide chains of lipopolysaccharides of P. aeruginosa O3 (Lányi), O25 (Wokatsch) and Fisher immunotypes 3 and 7" -
European Journal of Biochemistry 167 (1987) 549-561
O-specific polysaccharides, obtained on mild acid degradation of lipopolysacchrides of the serologically related strains Pseudomonas aeruginosa O3 (Lányi classification), O25 (Wokatsch classification) and immunotypes 3 and 7 (Fisher classification), are built up of trisaccharide repeating units involving 2-acetamido-2,6-dideoxy-D-galactose (N-acetyl-D-fucosamine), 2,3-diacetamido-2,3-dideoxy-D-mannuronic acid or 2,3-diacetamido-2,3-dideoxy-L-guluronic acid and 3-acetamidino-2-acetamido-2,3-dideoxy-D-mannuronic acid or 3-acetamidino-2-acetamido-2,3-dideoxy-L-guluronic acid. Lányi O3(a),3d,3f and Wokatsch O25 polysaccharides contain also O-acetyl groups. On the basis of solvolysis with anhydrous hydrogen fluoride, resulting in trisaccharide fragments with N-acetylfucosamine residue at the reducing terminus, chemical modifications of the acetamidino group (alkaline hydrolysis to the acetamido group or reductive deamination to the ethylamino group), as well as analysis by 1H-NMR (including nuclear Overhauser effect experiments) and 13C-NMR spectroscopy, and fast-atom bombardment mass spectrometry, it was concluded that the repeating units of the polysaccharides have the following structures: (Formula: see text) where HexNAcAmA = α-L-GulNAcAmA (approximately 70%) or β-D-ManNacAMA (approximately 30%). Lányi O3(a),3d,3f polysaccharide involves two types of repeating units, which differ from each other only in the configuration at C-5 of the 3-acetamidino-2-acetamido-2,3-dideoxyuronic acid residue. Lányi O3(a),3c,O3a,3d,3e and Fisher immunotypes 3 and 7 polysaccharides contain, together with the major repeating units shown above, a small proportion of units in which the derivative of α-L-guluronic acid is replaced by the corresponding β-D-manno isomer. The data obtained provide the opportunity to substantiate the serological interrelations between these strains of P. aeruginosa by the presence in the O-specific polysaccharides of common monosaccharides or disaccharide fragments. The distinctions between them stem from the presence or absence of the O-acetyl group, a different configuration of the glycosidic linkage of the N-acetylfucosamine residue and/or a different configuration at C-5 of one or both derivatives of diaminouronic acids.
NCBI PubMed ID: 3115777Journal NLM ID: 0107600Publisher: Oxford, UK: Blackwell Science Ltd. on behalf of the Federation of European Biochemical Societies
Institutions: N.D. Zelinsky Institute of Organic Chemistry, Academy of Sciences of the USSR, Moscow, Russia
Methods: 13C NMR, 1H NMR, FAB-MS
- Article ID: 3207
Knirel YA, Bystrova OV, Kocharova NA, Zähringer U, Pier GB "Conserved and variable structural features of the Pseudomonas aeruginosa lipopolysaccharide" -
Journal of Endotoxin Research 12(6) (2006) 324-336
The review is devoted to recent progress in the structural elucidation of the lipopolysaccharide of the bacterium Pseudomonas aeruginosa, including O-antigen biological repeats, core oligosaccharide, and lipid A. Data on biosynthesis, genetics and serology of the lipopolysaccharide isolated from various P. aeruginosa O-serogroups are discussed in relation to the chemical structures.
O-antigen, Pseudomonas aeruginosa, lipid A, core oligosaccharide, lipopolysaccharide structure, serologic activity
NCBI PubMed ID: 17254386Publication DOI: 10.1179/096805106X118906Journal NLM ID: 9433350Publisher: Maney Publishing
Correspondence: knirel@ioc.ac.ru
Institutions: N.D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia, Research Center Borstel, Leibniz-Center for Medicine and Biosciences, Borstel, Germany, Channing Laboratory, Brigham and Women’s Hospital, Harvard Medical School, Boston, Massachusetts, USA
Methods: NMR, chemical methods, MS, serological methods, genetic methods
- Article ID: 3391
Kaluzny K, Abeyrathne PD, Lam JS "Coexistence of Two Distinct Versions of O-Antigen Polymerase, Wzy-Alpha and Wzy-Beta, in Pseudomonas aeruginosa Serogroup O2 and Their Contributions to Cell Surface Diversity" -
Journal of Bacteriology 189(11) (2007) 4141-4152
Assembly of B-band LPS in Pseudomonas aeruginosa follows a Wzy-dependent pathway, requiring the O-antigen polymerase Wzy, and other proteins. The peptide sequences of the wzyα product from strains of serotypes O2, O5, and O16 are identical; but the O-units in O5 are α-glycosidic-linked, while those in O2 and O16 are β-linked. We hypothesized that a derivative of the D3 bacteriophage wzyβ is present in the chromosomes of O2 and O16, and that this gene is responsible for the β-linkage. By a combination of PCR and primer walking, wzyβ of both serotypes have been amplified and cloned. Both are identical, but only share 87.42% sequence identity with their xenolog in D3. A chromosomal knockout mutant of O16 wzyβ was made and it produces rough LPS devoid of B-band O antigen. The cloned wzyβ is capable of complementing the O16 wzyβ mutant, as well as cross-complementing a wzyα knockout mutant. However, in the latter case, the restored O-antigen was β-linked. Using RT-PCR, we showed that wzyα was transcribed in O2 and O16 strains and was functional since both of these genes could complement the wzyα mutant of O5. With the co-existence of wzyα and wzyβ in O2 and O16 and the B-band O polysaccharides in these being β-linked, we hypothesized that iap, an inhibitor of alpha-polymerase gene, must be present in these serotypes. Indeed, through PCR, TOPO cloning and nucleotide sequencing results, we verified the presence of iap in both O2 and O16 serotypes
serotype, Pseudomonas aeruginosa, O-polysaccharide, serogroup, sequencing, O-antigen polymerase, rough LPS
NCBI PubMed ID: 17384183Journal NLM ID: 2985120RPublisher: American Society for Microbiology
Correspondence: jlam@uoguelph.ca
Institutions: University of Guelph, Department of Molecular and Cellular Biology, Guelph, Ontario, Canada N1G 2W1
Methods: serological methods, genetic methods
- Article ID: 3711
King JD, Kocincova D, Westman EL, Lam JS "Lipopolysaccharide biosynthesis in Pseudomonas aeruginosa" -
Innate Immunity 15(5) (2009) 261-312
Pseudomonas aeruginosa causes serious nosocomial infections, and an important virulence factor produced by this organism is lipopolysaccharide (LPS). This review summarizes knowledge about biosynthesis of all three structural domains of LPS - lipid A, core oligosaccharide, and O polysaccharides. In addition, based on similarities with other bacterial species, this review proposes new hypothetical pathways for unstudied steps in the biosynthesis of P. aeruginosa LPS. Lipid A biosynthesis is discussed in relation to Escherichia coli and Salmonella, and the biosyntheses of core sugar precursors and core oligosaccharide are summarised. Pseudomonas aeruginosa attaches a Common Polysaccharide Antigen and O-Specific Antigen polysaccharides to lipid A-core. Both forms of O polysaccharide are discussed with respect to their independent synthesis mechanisms. Recent advances in understanding O-polysaccharide biosynthesis since the last major review on this subject, published nearly a decade ago, are highlighted. Since P. aeruginosa O polysaccharides contain unusual sugars, sugar-nucleotide biosynthesis pathways are reviewed in detail. Knowledge derived from detailed studies in the O5, O6 and O11 serotypes is applied to predict biosynthesis pathways of sugars in poorly-studied serotypes, especially O1, O4, and O13/O14. Although further work is required, a full understanding of LPS biosynthesis in P. aeruginosa is almost within reach.
Lipopolysaccharide, core, O-antigen, Pseudomonas aeruginosa, lipid A
NCBI PubMed ID: 19710102Publication DOI: 10.1177/1753425909106436Journal NLM ID: 101469670Publisher: Sage Publications
Correspondence: jlam@uoguelph.ca
Institutions: Department of Molecular and Cellular Biology, University of Guelph, Guelph, Ontario, Canada
- Article ID: 4062
Lam JS, Taylor VL, Islam ST, Hao Y, Kocincova D "Genetic and Functional Diversity of Pseudomonas aeruginosa Lipopolysaccharide" -
Frontiers in Microbiology 2 (2011) 118
Lipopolysccharide (LPS) is an integral component of the Pseudomonas aeruginosa cell envelope, occupying the outer leaflet of the outer membrane in this Gram-negative opportunistic pathogen. It is important for bacterium-host interactions and has been shown to be a major virulence factor for this organism. Structurally, P. aeruginosa LPS is composed of three domains, namely, lipid A, core oligosaccharide, and the distal O antigen (O-Ag). Most P. aeruginosa strains produce two distinct forms of O-Ag, one a homopolymer of D-rhamnose that is a common polysaccharide antigen (CPA, formerly termed A band), and the other a heteropolymer of three to five distinct (and often unique dideoxy) sugars in its repeat units, known as O-specific antigen (OSA, formerly termed B band). Compositional differences in the O units among the OSA from different strains form the basis of the International Antigenic Typing Scheme for classification via serotyping of different strains of P. aeruginosa. The focus of this review is to provide state-of-the-art knowledge on the genetic and resultant functional diversity of LPS produced by P. aeruginosa. The underlying factors contributing to this diversity will be thoroughly discussed and presented in the context of its contributions to host-pathogen interactions and the control/prevention of infection.
Lipopolysaccharide, biosynthesis, virulence, serotyping, bacteriophage, motility, seroconversion, nucleotide sugars
NCBI PubMed ID: 21687428Publication DOI: 10.3389/fmicb.2011.00118Journal NLM ID: 101548977Publisher: Lausanne: Frontiers Research Foundation
Correspondence: jlam@uoguelph.ca
Institutions: Department of Molecular and Cellular Biology, University of Guelph Guelph, ON, Canada
- Article ID: 4684
Islam ST, Lam JS "Synthesis of bacterial polysaccharides via the Wzx/Wzy-dependent pathway" -
Canadian Journal of Microbiology 60(11) (2014) 697-716
The surfaces of bacteria mediate a multitude of functions in the environment and in an infected host, including adhesion to both biotic and abiotic substrata, motility, immune system interaction and (or) activation, biofilm formation, and cell-cell communication, with many of these features directly influenced by cell-surface glycans. In both Gram-negative and Gram-positive bacteria, the majority of cell-surface polysaccharides are produced via the Wzx/Wzy-dependent assembly pathway; these glycans include heteropolymeric O-antigen, enterobacterial common antigen, exopolysaccharide, spore coat, and capsule in diverse bacteria. The key components of this assembly pathway are the integral inner membrane Wzx flippase, Wzy polymerase, and Wzz chain-length regulator proteins, which until recently have resisted detailed structural and functional characterization. In this review, we have provided a comprehensive synthesis of the latest structural and mechanistic data for each protein, as well as an examination of substrate specificity for each assembly step and complex formation between the constituent proteins. To complement the unprecedented explosion of genomic-sequencing data for bacteria, we have also highlighted both classical and state-of-the-art methods by which encoded Wzx, Wzy, and Wzz proteins can be reliably identified and annotated, using the model Gram-negative bacterium Pseudomonas aeruginosa as an example data set. Lastly, we outline future avenues of research, with the aim of stimulating researchers to take the next steps in investigating the function of, and interplay between, the constituents of this widespread assembly scheme.
Membrane Proteins, lipopolysaccharide (LPS), Wzx flippase, Wzy polymerase, Wzz polysaccharide copolymerase
NCBI PubMed ID: 25358682Publication DOI: 10.1139/cjm-2014-0595Journal NLM ID: 0372707Publisher: National Research Council of Canada
Correspondence: sislam@imm.cnrs.fr; jlam@uoguelph.ca
Institutions: Department of Molecular and Cellular Biology, University of Guelph, Guelph, ON N1G 2W1, Canada
Methods: SDS-PAGE, genetic methods, cloning
- Article ID: 4932
Taylor VL, Hoage JF, Thrane SW, Huszczynski SM, Jelsbak L, Lam JS "A Bacteriophage-Acquired O-Antigen Polymerase (Wzyb) from P. aeruginosa Serotype O16 Performs a Varied Mechanism Compared to Its Cognate Wzya" -
Frontiers in Microbiology 7 (2016) 393
Pseudomonas aeruginosa is a Gram-negative bacterium that produces highly varied lipopolysaccharide (LPS) structures. The O antigen (O-Ag) in the LPS is synthesized through the Wzx/Wzy-dependent pathway where lipid-linked O-Ag repeats are polymerized by Wzy. Horizontal-gene transfer has been associated with O-Ag diversity. The O-Ag present on the surface of serotypes O5 and O16, differ in the intra-molecular bonds, alpha and beta, respectively; the latter arose from the action of three genes in a serotype converting unit acquired from bacteriophage D3, including a beta-polymerase (Wzyβ). To further our understanding of O-polymerases, the inner membrane (IM) topology of Wzyβ was determined using a dual phoA-lacZα reporter system wherein random 3' gene truncations were localized to specific loci with respect to the IM by normalized reporter activities as determined through the ratio of alkaline phosphatase activity to β-galactosidase activity. The topology of Wzyβ developed through this approach was shown to contain two predominant periplasmic loops, PL3 (containing an RX10G motif) and PL4 (having an O-Ag ligase superfamily motif), associated with inverting glycosyltransferase reaction. Through site-directed mutagenesis and complementation assays, residues Arg(254), Arg(270), Arg(272), and His(300) were found to be essential for Wzyβ function. Additionally, like-charge substitutions, R254K and R270K, could not complement the wzyβ knockout, highlighting the essential guanidium side group of Arg residues. The O-Ag ligase domain is conserved among heterologous Wzy proteins that produce β-linked O-Ag repeat units. Taking advantage of the recently obtained whole-genome sequence of serotype O16 a candidate promoter was identified. Wzyβ under its native promoter was integrated in the PAO1 genome, which resulted in simultaneous production of α- and β-linked O-Ag. These observations established that members of Wzy-like family consistently exhibit a dual-periplasmic loops topology, and identifies motifs that are plausible to be involved in enzymatic activities. Based on these results, the phage-derived Wzyβ utilizes a different reaction mechanism in the P. aeruginosa host to avoid self-inhibition during serotype conversion.
Lipopolysaccharide, serotype, Pseudomonas aeruginosa, glycosyltransferase, bacteriophage, O-antigen biosynthesis, polymerase
NCBI PubMed ID: 27065964Publication DOI: 10.3389/fmicb.2016.00393Journal NLM ID: 101548977Publisher: Lausanne: Frontiers Research Foundation
Correspondence: jlam@uoguelph.ca
Institutions: Department of Molecular and Cellular Biology, University of Guelph Guelph, ON, Canada, Department of Systems Biology, Technical University of Denmark Kongens Lyngby, Denmark
Methods: PCR, SDS-PAGE, Western blotting, genetic methods, enzymatic assay
- Article ID: 5029
Olszak T, Shneider MM, Latka A, Maciejewska B, Browning C, Sycheva LV, Comer JE, Danis-Wlodarczyk K, Senchenkova SN, Shashkov AS, Gula G, Arabski M, Wasik S, Miroshnikov KA, Lavigne R, Leiman PG, Knirel YA, Drulis-Kawa Z "The O-specific polysaccharide lyase from the phage LKA1 tailspike reduces Pseudomonas virulence" -
Scientific Reports 7(1) (2017) 16302
Pseudomonas phage LKA1 of the subfamily Autographivirinae encodes a tailspike protein (LKA1gp49) which binds and cleaves B-band LPS (O-specific antigen, OSA) of Pseudomonas aeruginosa PAO1. The crystal structure of LKA1gp49 catalytic domain consists of a beta-helix, an insertion domain and a C-terminal discoidin-like domain. The putative substrate binding and processing site is located on the face of the beta-helix whereas the C-terminal domain is likely involved in carbohydrates binding. NMR spectroscopy and mass spectrometry analyses of degraded LPS (OSA) fragments show an O5 serotype-specific polysaccharide lyase specificity. LKA1gp49 reduces virulence in an in vivo Galleria mellonella infection model and sensitizes P. aeruginosa to serum complement activity. This enzyme causes biofilm degradation and does not affect the activity of ciprofloxacin and gentamicin. This is the first comprehensive report on LPS-degrading lyase derived from a Pseudomonas phage. Biological properties reveal a potential towards its applications in antimicrobial design and as a microbiological or biotechnological tool.
genetic, X-ray, O-specific polysaccharide, Enzymes, lyase, bacteriophage, Pseudomonas phage
Publication DOI: 10.1038/s41598-017-16411-4Journal NLM ID: 101563288Publisher: London: Nature Publishing Group
Correspondence: Zuzanna Drulis-Kawa
Institutions: N.D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia, Institute of Genetics and Microbiology, University of Wroclaw, Wroclaw, 51-148, Poland, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, Russia, University of Texas Medical Branch, Department of Biochemistry and Molecular Biology, Sealy Center for Structural Biology and Molecular Biophysics, Galveston, TX, 77555-0647, USA, Vertex Pharmaceuticals (Europe) Ltd, Abingdon, Oxfordshire, OX14 4RW, UK, Affinivax Inc., Cambridge, MA, USA, Laboratory of Gene Technology, KU Leuven, Leuven, 3001, Belgium, Department of Biochemistry and Genetics, Institute of Biology, The Jan Kochanowski University in Kielce, Kielce, 25-406, Poland, Department of Molecular Physics, Institute of Physics, The Jan Kochanowski University in Kielce, Kielce, 25-406, Poland
Methods: 13C NMR, 1H NMR, NMR-2D, X-ray, SDS-PAGE, sugar analysis, mild acid hydrolysis, biological assays, genetic methods, GPC, enzyme assay, cloning, crystallization, phage degradation, HR ESI-MS
- Article ID: 5061
Zheng H, Shashkov AS, Xiong Y, Naumenko OI, Wang H, Senchenkova SN, Wang J, Knirel YA "Structure and gene cluster of the O-antigen of Escherichia albertii O1 resembling the O-antigen of Pseudomonas aeruginosa O5" -
Carbohydrate Research (2017) 28-31
The O-specific polysaccharide (O-antigen) was obtained by mild acid degradation of the lipopolysaccharide of Escherichia albertii serotype O1 strain SP20140089 and studied by sugar analysis along with 1D and 2D 1H and 13C NMR spectroscopy. The following structure was established for the trisaccharide repeating unit of the O-polysaccharide: →4)-β-d-ManpNAc3NAcA-(1→4)-β-d-GlcpNAm3NAcA-(1→3)-α-d-GlcpNAc-(1→ where ManNAc3NAcA and GlcNAm3NAcA indicate 2,3-diacetamido-2,3-dideoxymannuronic acid and 2-acetimidoylamino-3-acetamido-2,3-dideoxyglucuronic acid, respectively. While showing some similarity with O-polysaccharide structures of a group of Pseudomonas aeruginosa serotypes (O2, O5, O16, O18, and O20), that of E. albertii O1 is unique among known bacterial polysaccharide structures. The gene cluster for biosynthesis of the O1-antigen was sequenced and functions of the genes were predicted by comparison with sequences in the available databases, including those involved in the synthesis of nucleotide precursors of 2,3-diamino-2,3-dideoxyhexuronic acid derivatives in P. aeruginosa O5.
Lipopolysaccharide, Pseudomonas aeruginosa, capsular polysaccharide, O-specific polysaccharide, bacterial polysaccharide structure, O-antigen gene cluster, Escherichia albertii
NCBI PubMed ID: 28494314Publication DOI: 10.1016/j.carres.2017.04.024Journal NLM ID: 0043535Publisher: Elsevier
Correspondence: Knirel YA
Institutions: N.D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia, Higher Chemical College of the Russian Academy of Sciences, D. I. Mendeleev University of Chemical Technology of Russia, Moscow, Russia, Zigong Center for Disease Control and Prevention, Zigong, Sichuan Province, China, State Key Laboratory of Infectious Disease Prevention and Control, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Changping, Beijing, China
Methods: 13C NMR, 1H NMR, NMR-2D, DNA sequencing, sugar analysis, acid hydrolysis, GPC, bioinformatic analysis
- Article ID: 5204
Qin C, Schumann B, Zou X, Pereira CL, Tian G, Xu J, Seeberger PH, Yin J "Total Synthesis of a Densely Functionalized Plesiomonas shigelloides Serotype 51 Aminoglycoside Trisaccharide Antigen" -
Journal of the American Chemical Society 140(8) (2018) 3120-3127
Plesiomonas shigelloides, a pathogen responsible for frequent outbreaks of severe travelers' diarrhea, causes grave extraintestinal infections. Sepsis and meningitis due to P. shigelloides are associated with a high mortality rate as antibiotic resistance increases and vaccines are not available. Carbohydrate antigens expressed by pathogens are often structurally unique and are targets for developing vaccines and diagnostics. Here, we report a total synthesis of the highly functionalized trisaccharide repeating unit 2 from P. shigelloides serotype 51 from three monosaccharides. A judicious choice of building blocks and reaction conditions allowed for the four amino groups adorning the sugar rings to be installed with two N-acetyl (Ac) groups, rare acetamidino (Am), and d-3-hydroxybutyryl (Hb) groups. The strategy for the differentiation of amino groups in trisaccharide 2 will serve well for the syntheses of other complex glycans.
synthesis, serotype, trisaccharide, vaccine, Plesiomonas shigelloides, total synthesis, carbohydrate chemistry
NCBI PubMed ID: 29377682Publication DOI: 10.1021/jacs.8b00148Journal NLM ID: 7503056Publisher: American Chemical Society
Correspondence: peter.seeberger@mpikg.mpg.de; jianyin@jiangnan.edu.cn
Institutions: Department of Biomolecular Systems, Max Planck Institute of Colloids and Interfaces , Am Muhlenberg 1, 14476 Potsdam, Germany, Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University , Wuxi, Jiangsu Province 214122, P.R. China
Methods: 13C NMR, 1H NMR, NMR-2D, chemical synthesis, chemical methods, glycosylation
- Article ID: 5760
Dobrochaeva K, Khasbiulina N, Shilova N, Antipova N, Obukhova P, Galanina O, Blixt O, Kunz H, Filatov A, Knirel Y, Le Pendu J, Khaidukov S, Bovin N "Specificity of human natural antibodies referred to as anti-Tn" -
Molecular Immunology 120 (2020) 74-82
To understand the role of human natural IgM known as antibodies against the carbohydrate epitope Tn, the antibodies were isolated using GalNAcα-Sepharose affinity chromatography, and their specificity was profiled using microarrays (a glycan array printed with oligosaccharides and bacterial polysaccharides, as well as a glycopeptide array), flow cytometry, and inhibition ELISA. The antibodies bound a restricted number of GalNAcα-terminated oligosaccharides better than the parent monosaccharide, e.g., 6-O-Su-GalNAcα and GalNAcα1-3Galβ1-3(4)GlcNAcβ. The binding with several bacterial polysaccharides that have no structural resemblance to the affinity ligand GalNAcα was quite unexpected. Given that GalNAcα is considered the key fragment of the Tn antigen, it is surprising that these antibodies bind weakly GalNAcα-OSer and do not bind a wide variety of GalNAcα-OSer/Thr-containing mucin glycopeptides. At the same time, we have observed specific binding to cells having Tn-positive glycoproteins containing similar glycopeptide motifs in a conformationally rigid macromolecule. Thus, specific recognition of the Tn antigen apparently requires that the naturally occurring "anti-Tn" IgM recognize a complex epitope comprising the GalNAcα as an essential component and a fairly long amino acid sequence where the amino acids adjacent to GalNAcα do not contact the antibody paratope; i.e., the antibodies recognize a spatial epitope or a molecular pattern rather than a classical continuous sequence. In addition, we have not found any increase in the binding of natural antibodies when GalNAcα residues were clustered. These results may help in further development of anticancer vaccines based on synthetic Tn constructs.
cancer, glycans, natural antibodies, anti-glycan antibodies, Tn antigen
NCBI PubMed ID: 32087569Publication DOI: 10.1016/j.molimm.2020.02.005Journal NLM ID: 7905289Publisher: Elsevier
Correspondence: professorbovin@yandex.ru
Institutions: Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 16/10 Miklukho-Maklaya, Moscow, Russian Federation, Semiotik LLC, 16/10 Miklukho-Maklaya, Moscow, Russian Federation, National Medical Research Center for Obstetrics, Gynecology and Perinatology Named after Academician V.I. Kulakov of the Ministry of Healthcare of Russian Federation, Moscow, Russian Federation, National Research University Higher School of Economics, Moscow, Russian Federation, Department of Chemistry, Chemical Biology, University of Copenhagen, Thorvaldsensvej 40, 1871 Frederiksberg C, Denmark, Institut Fur Organische Chemie, Johannes Gutenberg-Universitat Mainz, Duesbergweg 10-14, D-55128, Mainz, Germany, Institute of Immunology, Federal Medical-Biological Agency of Russia, Moscow, Russian Federation, University of Nantes, Inserm, U892 IRT UN, 8 Quai MonCousu, BP70721 Nantes, FR 44007, France
Methods: ELISA, affinity chromatography, flow cytometry analysis, printed glycan array (PGA) analysis, FACS assay
- Article ID: 5835
Richard G, MacKenzie CR, Henry KA, Vinogradov E, Hall JC, Hussack G "Antibody Binding to the O-Specific Antigen of Pseudomonas aeruginosa O6 Inhibits Cell Growth" -
Antimicrobial Agents and Chemotherapy 64(4) (2020) e02168
Pseudomonas aeruginosa is an opportunistic pathogen that is inherently resistant to many antibiotics and represents an increasing threat due to the emergence of drug-resistant strains. There is a pressing need to develop innovative antimicrobials against this pathogen. In this study we identified the O-specific antigen (OSA) of P. aeruginosa serotype O6 as a novel target for therapeutic intervention. Binding of monoclonal antibodies and antigen-binding fragments therefrom to O6 OSA leads to rapid outer membrane destabilization and inhibition of cell growth. The antimicrobial effect correlated directly with antibody affinity. Antibody binding to the O-antigen of a second lipopolysaccharide type present in P. aeruginosa or to the LPS core did not affect cell viability. Atomic force microscopy showed that antibody binding to OSA resulted in early flagellum loss, formation of membrane blebs, and eventually complete outer membrane loss. We hypothesize that antibody binding to OSA disrupts a key interaction in the P. aeruginosa outer membrane.
Lipopolysaccharide, LPS, AFM, atomic force microscopy, antibacterial antibodies, O-specific antigen, outer membrane disruption
NCBI PubMed ID: 32015038Publication DOI: 10.1128/AAC.02168-19Journal NLM ID: 0315061Correspondence: Greg.Hussack@nrc-cnrc.gc.ca
Institutions: School of Environmental Sciences, University of Guelph, Guelph, ON, Canada N1G 2W1, Department of Biochemistry, Microbiology and Immunology, University of Ottawa, Ottawa, ON, Canada K1H 8M5, Human Health Therapeutics Research Centre, National Research Council Canada, Ottawa, ON, Canada K1A 0R6
Methods: ELISA, serological methods, SPR, AFM, antibody sequence analyses
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8. Compound ID: 965
|
-4)-b-D-ManpNAc3NAmA-(1-4)-a-L-GulpNAc3NAcA-(1-3)-a-D-FucpNAc-(1- |
Show graphically |
Structure type: polymer chemical repeating unit
Compound class: O-polysaccharide, O-antigen
The structure is contained in the following publication(s):
- Article ID: 254
Hatano K, Pier GB "Complex serology and immune response of mice to variant high-molecular-weight O polysaccharide isolated from Pseudomonas aeruginosa serogroup O2 strains" -
Infection and Immunity 66(8) (1998) 3719-3726
The O antigen of the Pseudomonas aeruginosa lipopolysaccharide is the optimal target for protective antibodies, but the unusual and complex nature of their sugar substituents has made it difficult to define the range of these structures needed in an effective vaccine. Most clinical isolates of P. aeruginosa can be classified into 10 O-antigen serogroups, but slight chemical differences among O polysaccharides within a serogroup give rise to subtype epitopes. These epitopes could impact the reactivity of O-antigen-specific antibodies, as well as the susceptibility of a target strain to protective, opsonic antibodies. To define parameters of serogroup and subtype-epitope immunogenicity, antigenicity, and surface expression on P. aeruginosa cells, we prepared high-molecular-weight O-polysaccharide vaccines from strains of P. aeruginosa serogroup O2, for which eight structurally variant O antigens expressing six defined subtype epitopes (O2a to O2f) have been identified. A complex pattern of immune responses to these antigens was observed following vaccination of mice. The high-molecular-weight O polysaccharides were generally more immunogenic at low doses (1 and 10 microg) than at a high dose (50 microg) and usually elicited antibodies that opsonized the homologous strain for phagocytic killing. Some of the individual polysaccharides elicited cross-opsonic antibodies to a variable number of strains that express all of the defined serogroup O2 subtype epitopes. Combination into one vaccine of two antigens that individually elicited cross-reactive opsonic antibodies to most members of the O2 serogroup inhibited, instead of enhanced, the production of antibodies broadly reactive with most serogroup O2 subtype strains. Thus, immune responses to P. aeruginosa O antigens may be restricted to a limited range of epitopes on structurally complex O antigens, and combining multiple related antigens into a single vaccine formulation may inhibit the production of those antibodies best able to protect against most P. aeruginosa strains within a given O-antigen serogroup.
Lipopolysaccharide, Pseudomonas, Pseudomonas aeruginosa, O-polysaccharide, serology, serogroup, immune response
NCBI PubMed ID: 9673254Journal NLM ID: 0246127Publisher: American Society for Microbiology
Correspondence: gpier@channing.harvard.edu
Institutions: Charming Laboratory, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115-5804., Charming Laboratory, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School,Boston, Massachusetts 02115-5804.
- Article ID: 1412
de Kievit TR, Staples T, Lam JS "Pseudomonas aeruginosa rfc genes of serotypes O2 and O5 could complement O-polymerase-deficient semi-rough mutants of either serotype" -
FEMS Microbiology Reviews 147(2) (1997) 251-257
Using a gene-replacement strategy and a mutated copy of the Pseudomonas aeruginosa O5 rfc gene, we were able to generate a rfc mutant in P. aeruginosa serotype O2. This mutant, which exhibits the semi-rough (SR) LPS phenotype, was used to isolate the O2 rfc gene. Mobilization of the O2 and O5 rfc genes into SR mutants of the heterologous serotype resulted in 'cross-polymerization' of O-repeat units, indicating that the genes are functionally exchangeable. Analysis of the nucleotide sequence of the rfc genes revealed that the two Rfc proteins are identical. The results of this study have enabled us to propose the linkage catalyzed by the O5 O-polymerase enzyme.
Lipopolysaccharide, LPS, gene, serotype, Pseudomonas, Pseudomonas aeruginosa, mutant, mutants, Serotypes, O-antigen polymerase, rfc, complement, semirough
NCBI PubMed ID: 9119201Journal NLM ID: 8902526Publisher: Oxford University Press
Correspondence: jlam@micro.uoguelph.ca
Institutions: Department of Microbiology, University of Guelph, Guelph, Canada
- Article ID: 1778
Knirel YA, Kochetkov NK "The structure of lipopolysaccharides of gram-negative bacteria. III. The structure of O-antigens: A review" -
Biochemistry (Moscow) 59(12) (1994) 1325-1383
This review summarizes data on the composition and structure of the O-antigens, the polysaccharide chains of the outer-membrane lipopolysaccharides (LPS) of Gram-negative bacteria defining the immunospecificity of these microbial cells. Special reference is given to some structural features of the O-antigens, such as the presence of unique monosaccharides and noncarbohydrate components, masked regularity, and the occurrence in one microorganism of LPS with structurally different polysaccharide chains. Antigenic relationships between microorganisms belonging to different taxonomic groups are discussed.
structure, O-antigen, chemical composition, bacterial lipopolysaccharides, Salmonella livingstone C1
NCBI PubMed ID: 7533007Journal NLM ID: 0376536Publisher: Nauka/Interperiodica
Institutions: Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia
- Article ID: 2409
Knirel YA, Paramonov NA, Vinogradov EV, Shashkov AS, Dmitriev BA, Kochetkov NK, Kholodkova EV, Stanislavsky ES "Somatic antigens of Pseudomonas aeruginosa. The structure of O-specific polysaccharide chains of lipopolysaccharides of P. aeruginosa O3 (Lányi), O25 (Wokatsch) and Fisher immunotypes 3 and 7" -
European Journal of Biochemistry 167 (1987) 549-561
O-specific polysaccharides, obtained on mild acid degradation of lipopolysacchrides of the serologically related strains Pseudomonas aeruginosa O3 (Lányi classification), O25 (Wokatsch classification) and immunotypes 3 and 7 (Fisher classification), are built up of trisaccharide repeating units involving 2-acetamido-2,6-dideoxy-D-galactose (N-acetyl-D-fucosamine), 2,3-diacetamido-2,3-dideoxy-D-mannuronic acid or 2,3-diacetamido-2,3-dideoxy-L-guluronic acid and 3-acetamidino-2-acetamido-2,3-dideoxy-D-mannuronic acid or 3-acetamidino-2-acetamido-2,3-dideoxy-L-guluronic acid. Lányi O3(a),3d,3f and Wokatsch O25 polysaccharides contain also O-acetyl groups. On the basis of solvolysis with anhydrous hydrogen fluoride, resulting in trisaccharide fragments with N-acetylfucosamine residue at the reducing terminus, chemical modifications of the acetamidino group (alkaline hydrolysis to the acetamido group or reductive deamination to the ethylamino group), as well as analysis by 1H-NMR (including nuclear Overhauser effect experiments) and 13C-NMR spectroscopy, and fast-atom bombardment mass spectrometry, it was concluded that the repeating units of the polysaccharides have the following structures: (Formula: see text) where HexNAcAmA = α-L-GulNAcAmA (approximately 70%) or β-D-ManNacAMA (approximately 30%). Lányi O3(a),3d,3f polysaccharide involves two types of repeating units, which differ from each other only in the configuration at C-5 of the 3-acetamidino-2-acetamido-2,3-dideoxyuronic acid residue. Lányi O3(a),3c,O3a,3d,3e and Fisher immunotypes 3 and 7 polysaccharides contain, together with the major repeating units shown above, a small proportion of units in which the derivative of α-L-guluronic acid is replaced by the corresponding β-D-manno isomer. The data obtained provide the opportunity to substantiate the serological interrelations between these strains of P. aeruginosa by the presence in the O-specific polysaccharides of common monosaccharides or disaccharide fragments. The distinctions between them stem from the presence or absence of the O-acetyl group, a different configuration of the glycosidic linkage of the N-acetylfucosamine residue and/or a different configuration at C-5 of one or both derivatives of diaminouronic acids.
NCBI PubMed ID: 3115777Journal NLM ID: 0107600Publisher: Oxford, UK: Blackwell Science Ltd. on behalf of the Federation of European Biochemical Societies
Institutions: N.D. Zelinsky Institute of Organic Chemistry, Academy of Sciences of the USSR, Moscow, Russia
Methods: 13C NMR, 1H NMR, FAB-MS
- Article ID: 3207
Knirel YA, Bystrova OV, Kocharova NA, Zähringer U, Pier GB "Conserved and variable structural features of the Pseudomonas aeruginosa lipopolysaccharide" -
Journal of Endotoxin Research 12(6) (2006) 324-336
The review is devoted to recent progress in the structural elucidation of the lipopolysaccharide of the bacterium Pseudomonas aeruginosa, including O-antigen biological repeats, core oligosaccharide, and lipid A. Data on biosynthesis, genetics and serology of the lipopolysaccharide isolated from various P. aeruginosa O-serogroups are discussed in relation to the chemical structures.
O-antigen, Pseudomonas aeruginosa, lipid A, core oligosaccharide, lipopolysaccharide structure, serologic activity
NCBI PubMed ID: 17254386Publication DOI: 10.1179/096805106X118906Journal NLM ID: 9433350Publisher: Maney Publishing
Correspondence: knirel@ioc.ac.ru
Institutions: N.D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia, Research Center Borstel, Leibniz-Center for Medicine and Biosciences, Borstel, Germany, Channing Laboratory, Brigham and Women’s Hospital, Harvard Medical School, Boston, Massachusetts, USA
Methods: NMR, chemical methods, MS, serological methods, genetic methods
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9. Compound ID: 966
|
-4)-b-D-ManpNAc3NAmA-(1-4)-b-D-ManpNAc3NAcA-(1-3)-a-D-FucpNAc4Ac-(1- |
Show graphically |
Structure type: polymer chemical repeating unit
Compound class: O-polysaccharide, O-antigen
Contained glycoepitopes: IEDB_2275410
The structure is contained in the following publication(s):
- Article ID: 254
Hatano K, Pier GB "Complex serology and immune response of mice to variant high-molecular-weight O polysaccharide isolated from Pseudomonas aeruginosa serogroup O2 strains" -
Infection and Immunity 66(8) (1998) 3719-3726
The O antigen of the Pseudomonas aeruginosa lipopolysaccharide is the optimal target for protective antibodies, but the unusual and complex nature of their sugar substituents has made it difficult to define the range of these structures needed in an effective vaccine. Most clinical isolates of P. aeruginosa can be classified into 10 O-antigen serogroups, but slight chemical differences among O polysaccharides within a serogroup give rise to subtype epitopes. These epitopes could impact the reactivity of O-antigen-specific antibodies, as well as the susceptibility of a target strain to protective, opsonic antibodies. To define parameters of serogroup and subtype-epitope immunogenicity, antigenicity, and surface expression on P. aeruginosa cells, we prepared high-molecular-weight O-polysaccharide vaccines from strains of P. aeruginosa serogroup O2, for which eight structurally variant O antigens expressing six defined subtype epitopes (O2a to O2f) have been identified. A complex pattern of immune responses to these antigens was observed following vaccination of mice. The high-molecular-weight O polysaccharides were generally more immunogenic at low doses (1 and 10 microg) than at a high dose (50 microg) and usually elicited antibodies that opsonized the homologous strain for phagocytic killing. Some of the individual polysaccharides elicited cross-opsonic antibodies to a variable number of strains that express all of the defined serogroup O2 subtype epitopes. Combination into one vaccine of two antigens that individually elicited cross-reactive opsonic antibodies to most members of the O2 serogroup inhibited, instead of enhanced, the production of antibodies broadly reactive with most serogroup O2 subtype strains. Thus, immune responses to P. aeruginosa O antigens may be restricted to a limited range of epitopes on structurally complex O antigens, and combining multiple related antigens into a single vaccine formulation may inhibit the production of those antibodies best able to protect against most P. aeruginosa strains within a given O-antigen serogroup.
Lipopolysaccharide, Pseudomonas, Pseudomonas aeruginosa, O-polysaccharide, serology, serogroup, immune response
NCBI PubMed ID: 9673254Journal NLM ID: 0246127Publisher: American Society for Microbiology
Correspondence: gpier@channing.harvard.edu
Institutions: Charming Laboratory, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115-5804., Charming Laboratory, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School,Boston, Massachusetts 02115-5804.
- Article ID: 337
Newton GJ, Daniels C, Burrows LL, Kropinski AM, Clarke AJ, Lam JS "Three-component-mediated serotype conversion in Pseudomonas aeruginosa by bacteriophage D3" -
Molecular Microbiology 39(5) (2001) 1237-1247
Bacteriophage D3 is capable of lysogenizing Pseudomonas aeruginosa PAO1 (serotype O5), converting the O-antigen from O5 to O16 and O-acetylating the N-acetylfucosamine moiety. To investigate the mechanism of lysogenic conversion, a 3.6 kb fragment from the D3 genome was isolated capable of mediating serotypic conversion identical to the D3 lysogen strain (AK1380). The PAO1 transformants containing this 3.6 kb of D3 DNA exhibited identical lipopolysaccharide (LPS) banding patterns to serotype O16 in silver-stained SDS-PAGE gels and displayed reactivity to an antibody specific for O-acetyl groups. Further analysis led to the identification of three open reading frames (ORFs) required for serotype conversion: an α-polymerase inhibitor (iap); an O-acetylase (oac); and a β-polymerase (wzyβ). The α-polymerase inhibitor (Iap) is capable of inhibiting the assembly of the serotype-specific O5 B-band LPS and allows the phage-encoded β-polymerase (Wzyβ) to form new β-linked B-band LPS. The D3 phage also alters the LPS by the addition of O-acetyl groups to the FucNAc residue in the O-antigen repeat unit by the action of the D3 O-acetylase (Oac). These three components form a simple yet elegant system by which bacteriophage D3 is capable of altering the surface of P. aeruginosa PAO1
serotype, Pseudomonas, Pseudomonas aeruginosa, bacteriophage, conversion, serotype conversion
NCBI PubMed ID: 11251840Journal NLM ID: 8712028Publisher: Blackwell Publishing
Correspondence: jlam@uoguelph.ca
Institutions: Department of Microbiology, University of Guelph, Guelph, Ontario, N1G 2W1, Canada, The Centre for Infection and Biomaterials Research, Toronto General Hospital, Toronto, Ontario, M5G 2C4, Canada, Department of Microbiology and Immunology, Queen's University, Kingston, Ontario, K7L 3N6, Canada
Methods: PCR, SDS-PAGE, DNA techniques, Western blotting, quantification of O-acetylation
- Article ID: 1778
Knirel YA, Kochetkov NK "The structure of lipopolysaccharides of gram-negative bacteria. III. The structure of O-antigens: A review" -
Biochemistry (Moscow) 59(12) (1994) 1325-1383
This review summarizes data on the composition and structure of the O-antigens, the polysaccharide chains of the outer-membrane lipopolysaccharides (LPS) of Gram-negative bacteria defining the immunospecificity of these microbial cells. Special reference is given to some structural features of the O-antigens, such as the presence of unique monosaccharides and noncarbohydrate components, masked regularity, and the occurrence in one microorganism of LPS with structurally different polysaccharide chains. Antigenic relationships between microorganisms belonging to different taxonomic groups are discussed.
structure, O-antigen, chemical composition, bacterial lipopolysaccharides, Salmonella livingstone C1
NCBI PubMed ID: 7533007Journal NLM ID: 0376536Publisher: Nauka/Interperiodica
Institutions: Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia
- Article ID: 1842
Knirel YA, Paramonov NA, Vinogradov EV, Shashkov AS, Dmitriev BA, Kochetkov NK, Stanislavsky ES "Antigenic polysaccharides of bacteria. 20. Structure of the O-specific polysaccharide chain of P. aeruginosa O3(a),3d,3f (Lányi) lipopolysaccharide" -
Bioorganicheskaya Khimia = Bioorganic Chemistry [Russian] 12 (1986) 1649-1657
Mild acid degradation of lipopolysaccharide from Pseudomonas aeruginosa O(3a), 3d, 3f (Lányi classification) afforded O-specific polysaccharide containing N-acetyl-D-fucosamine, 2,3-diacetamido-2,3-dideoxy-D-mannuronic acid, 3-acetamidino-2-acetamido-2,3-dideoxy-L-guluronic and D-mannuronic acid as well as O-acetyl groups. On the basis of O-deacetylation, selective cleavage with anhydrous fluoride, chemical transformation of the oligosaccharides obtained (hydrolysis or reductive deamination of the acetamidino group into acetamido or ethylamino group, respectively) and analysis by 13C NMR spectroscopy, it was concluded that the polysaccharide is built up mainly by trisaccharide repeating units of types A and B in the ratio approximately 2:1: (Formula: see text). The units of both types most probably enter the same polymeric chain. If so, such a hybrid structure can be accounted for by incompleteness of epimerization at C5 of the acetamidino derivative of mannuronic acid at the polymer level in the course of biosynthesis of this polysaccharide.
NCBI PubMed ID: 2434103Journal NLM ID: 7804941Publisher: Moskva: Nauka
Institutions: N.D. Zelinsky Institute of Organic Chemistry, Academy of Sciences of the USSR, Moscow, Russia
Methods: 13C NMR, 1H NMR
- Article ID: 2409
Knirel YA, Paramonov NA, Vinogradov EV, Shashkov AS, Dmitriev BA, Kochetkov NK, Kholodkova EV, Stanislavsky ES "Somatic antigens of Pseudomonas aeruginosa. The structure of O-specific polysaccharide chains of lipopolysaccharides of P. aeruginosa O3 (Lányi), O25 (Wokatsch) and Fisher immunotypes 3 and 7" -
European Journal of Biochemistry 167 (1987) 549-561
O-specific polysaccharides, obtained on mild acid degradation of lipopolysacchrides of the serologically related strains Pseudomonas aeruginosa O3 (Lányi classification), O25 (Wokatsch classification) and immunotypes 3 and 7 (Fisher classification), are built up of trisaccharide repeating units involving 2-acetamido-2,6-dideoxy-D-galactose (N-acetyl-D-fucosamine), 2,3-diacetamido-2,3-dideoxy-D-mannuronic acid or 2,3-diacetamido-2,3-dideoxy-L-guluronic acid and 3-acetamidino-2-acetamido-2,3-dideoxy-D-mannuronic acid or 3-acetamidino-2-acetamido-2,3-dideoxy-L-guluronic acid. Lányi O3(a),3d,3f and Wokatsch O25 polysaccharides contain also O-acetyl groups. On the basis of solvolysis with anhydrous hydrogen fluoride, resulting in trisaccharide fragments with N-acetylfucosamine residue at the reducing terminus, chemical modifications of the acetamidino group (alkaline hydrolysis to the acetamido group or reductive deamination to the ethylamino group), as well as analysis by 1H-NMR (including nuclear Overhauser effect experiments) and 13C-NMR spectroscopy, and fast-atom bombardment mass spectrometry, it was concluded that the repeating units of the polysaccharides have the following structures: (Formula: see text) where HexNAcAmA = α-L-GulNAcAmA (approximately 70%) or β-D-ManNacAMA (approximately 30%). Lányi O3(a),3d,3f polysaccharide involves two types of repeating units, which differ from each other only in the configuration at C-5 of the 3-acetamidino-2-acetamido-2,3-dideoxyuronic acid residue. Lányi O3(a),3c,O3a,3d,3e and Fisher immunotypes 3 and 7 polysaccharides contain, together with the major repeating units shown above, a small proportion of units in which the derivative of α-L-guluronic acid is replaced by the corresponding β-D-manno isomer. The data obtained provide the opportunity to substantiate the serological interrelations between these strains of P. aeruginosa by the presence in the O-specific polysaccharides of common monosaccharides or disaccharide fragments. The distinctions between them stem from the presence or absence of the O-acetyl group, a different configuration of the glycosidic linkage of the N-acetylfucosamine residue and/or a different configuration at C-5 of one or both derivatives of diaminouronic acids.
NCBI PubMed ID: 3115777Journal NLM ID: 0107600Publisher: Oxford, UK: Blackwell Science Ltd. on behalf of the Federation of European Biochemical Societies
Institutions: N.D. Zelinsky Institute of Organic Chemistry, Academy of Sciences of the USSR, Moscow, Russia
Methods: 13C NMR, 1H NMR, FAB-MS
- Article ID: 4932
Taylor VL, Hoage JF, Thrane SW, Huszczynski SM, Jelsbak L, Lam JS "A Bacteriophage-Acquired O-Antigen Polymerase (Wzyb) from P. aeruginosa Serotype O16 Performs a Varied Mechanism Compared to Its Cognate Wzya" -
Frontiers in Microbiology 7 (2016) 393
Pseudomonas aeruginosa is a Gram-negative bacterium that produces highly varied lipopolysaccharide (LPS) structures. The O antigen (O-Ag) in the LPS is synthesized through the Wzx/Wzy-dependent pathway where lipid-linked O-Ag repeats are polymerized by Wzy. Horizontal-gene transfer has been associated with O-Ag diversity. The O-Ag present on the surface of serotypes O5 and O16, differ in the intra-molecular bonds, alpha and beta, respectively; the latter arose from the action of three genes in a serotype converting unit acquired from bacteriophage D3, including a beta-polymerase (Wzyβ). To further our understanding of O-polymerases, the inner membrane (IM) topology of Wzyβ was determined using a dual phoA-lacZα reporter system wherein random 3' gene truncations were localized to specific loci with respect to the IM by normalized reporter activities as determined through the ratio of alkaline phosphatase activity to β-galactosidase activity. The topology of Wzyβ developed through this approach was shown to contain two predominant periplasmic loops, PL3 (containing an RX10G motif) and PL4 (having an O-Ag ligase superfamily motif), associated with inverting glycosyltransferase reaction. Through site-directed mutagenesis and complementation assays, residues Arg(254), Arg(270), Arg(272), and His(300) were found to be essential for Wzyβ function. Additionally, like-charge substitutions, R254K and R270K, could not complement the wzyβ knockout, highlighting the essential guanidium side group of Arg residues. The O-Ag ligase domain is conserved among heterologous Wzy proteins that produce β-linked O-Ag repeat units. Taking advantage of the recently obtained whole-genome sequence of serotype O16 a candidate promoter was identified. Wzyβ under its native promoter was integrated in the PAO1 genome, which resulted in simultaneous production of α- and β-linked O-Ag. These observations established that members of Wzy-like family consistently exhibit a dual-periplasmic loops topology, and identifies motifs that are plausible to be involved in enzymatic activities. Based on these results, the phage-derived Wzyβ utilizes a different reaction mechanism in the P. aeruginosa host to avoid self-inhibition during serotype conversion.
Lipopolysaccharide, serotype, Pseudomonas aeruginosa, glycosyltransferase, bacteriophage, O-antigen biosynthesis, polymerase
NCBI PubMed ID: 27065964Publication DOI: 10.3389/fmicb.2016.00393Journal NLM ID: 101548977Publisher: Lausanne: Frontiers Research Foundation
Correspondence: jlam@uoguelph.ca
Institutions: Department of Molecular and Cellular Biology, University of Guelph Guelph, ON, Canada, Department of Systems Biology, Technical University of Denmark Kongens Lyngby, Denmark
Methods: PCR, SDS-PAGE, Western blotting, genetic methods, enzymatic assay
- Article ID: 5760
Dobrochaeva K, Khasbiulina N, Shilova N, Antipova N, Obukhova P, Galanina O, Blixt O, Kunz H, Filatov A, Knirel Y, Le Pendu J, Khaidukov S, Bovin N "Specificity of human natural antibodies referred to as anti-Tn" -
Molecular Immunology 120 (2020) 74-82
To understand the role of human natural IgM known as antibodies against the carbohydrate epitope Tn, the antibodies were isolated using GalNAcα-Sepharose affinity chromatography, and their specificity was profiled using microarrays (a glycan array printed with oligosaccharides and bacterial polysaccharides, as well as a glycopeptide array), flow cytometry, and inhibition ELISA. The antibodies bound a restricted number of GalNAcα-terminated oligosaccharides better than the parent monosaccharide, e.g., 6-O-Su-GalNAcα and GalNAcα1-3Galβ1-3(4)GlcNAcβ. The binding with several bacterial polysaccharides that have no structural resemblance to the affinity ligand GalNAcα was quite unexpected. Given that GalNAcα is considered the key fragment of the Tn antigen, it is surprising that these antibodies bind weakly GalNAcα-OSer and do not bind a wide variety of GalNAcα-OSer/Thr-containing mucin glycopeptides. At the same time, we have observed specific binding to cells having Tn-positive glycoproteins containing similar glycopeptide motifs in a conformationally rigid macromolecule. Thus, specific recognition of the Tn antigen apparently requires that the naturally occurring "anti-Tn" IgM recognize a complex epitope comprising the GalNAcα as an essential component and a fairly long amino acid sequence where the amino acids adjacent to GalNAcα do not contact the antibody paratope; i.e., the antibodies recognize a spatial epitope or a molecular pattern rather than a classical continuous sequence. In addition, we have not found any increase in the binding of natural antibodies when GalNAcα residues were clustered. These results may help in further development of anticancer vaccines based on synthetic Tn constructs.
cancer, glycans, natural antibodies, anti-glycan antibodies, Tn antigen
NCBI PubMed ID: 32087569Publication DOI: 10.1016/j.molimm.2020.02.005Journal NLM ID: 7905289Publisher: Elsevier
Correspondence: professorbovin@yandex.ru
Institutions: Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 16/10 Miklukho-Maklaya, Moscow, Russian Federation, Semiotik LLC, 16/10 Miklukho-Maklaya, Moscow, Russian Federation, National Medical Research Center for Obstetrics, Gynecology and Perinatology Named after Academician V.I. Kulakov of the Ministry of Healthcare of Russian Federation, Moscow, Russian Federation, National Research University Higher School of Economics, Moscow, Russian Federation, Department of Chemistry, Chemical Biology, University of Copenhagen, Thorvaldsensvej 40, 1871 Frederiksberg C, Denmark, Institut Fur Organische Chemie, Johannes Gutenberg-Universitat Mainz, Duesbergweg 10-14, D-55128, Mainz, Germany, Institute of Immunology, Federal Medical-Biological Agency of Russia, Moscow, Russian Federation, University of Nantes, Inserm, U892 IRT UN, 8 Quai MonCousu, BP70721 Nantes, FR 44007, France
Methods: ELISA, affinity chromatography, flow cytometry analysis, printed glycan array (PGA) analysis, FACS assay
Expand this compound
Collapse this compound
10. Compound ID: 967
|
-4)-a-L-GulpNAc3NAmA-(1-4)-b-D-ManpNAc3NAcA-(1-3)-a-D-FucpNAc4Ac-(1- |
Show graphically |
Structure type: polymer chemical repeating unit
Compound class: O-polysaccharide, O-antigen
The structure is contained in the following publication(s):
- Article ID: 254
Hatano K, Pier GB "Complex serology and immune response of mice to variant high-molecular-weight O polysaccharide isolated from Pseudomonas aeruginosa serogroup O2 strains" -
Infection and Immunity 66(8) (1998) 3719-3726
The O antigen of the Pseudomonas aeruginosa lipopolysaccharide is the optimal target for protective antibodies, but the unusual and complex nature of their sugar substituents has made it difficult to define the range of these structures needed in an effective vaccine. Most clinical isolates of P. aeruginosa can be classified into 10 O-antigen serogroups, but slight chemical differences among O polysaccharides within a serogroup give rise to subtype epitopes. These epitopes could impact the reactivity of O-antigen-specific antibodies, as well as the susceptibility of a target strain to protective, opsonic antibodies. To define parameters of serogroup and subtype-epitope immunogenicity, antigenicity, and surface expression on P. aeruginosa cells, we prepared high-molecular-weight O-polysaccharide vaccines from strains of P. aeruginosa serogroup O2, for which eight structurally variant O antigens expressing six defined subtype epitopes (O2a to O2f) have been identified. A complex pattern of immune responses to these antigens was observed following vaccination of mice. The high-molecular-weight O polysaccharides were generally more immunogenic at low doses (1 and 10 microg) than at a high dose (50 microg) and usually elicited antibodies that opsonized the homologous strain for phagocytic killing. Some of the individual polysaccharides elicited cross-opsonic antibodies to a variable number of strains that express all of the defined serogroup O2 subtype epitopes. Combination into one vaccine of two antigens that individually elicited cross-reactive opsonic antibodies to most members of the O2 serogroup inhibited, instead of enhanced, the production of antibodies broadly reactive with most serogroup O2 subtype strains. Thus, immune responses to P. aeruginosa O antigens may be restricted to a limited range of epitopes on structurally complex O antigens, and combining multiple related antigens into a single vaccine formulation may inhibit the production of those antibodies best able to protect against most P. aeruginosa strains within a given O-antigen serogroup.
Lipopolysaccharide, Pseudomonas, Pseudomonas aeruginosa, O-polysaccharide, serology, serogroup, immune response
NCBI PubMed ID: 9673254Journal NLM ID: 0246127Publisher: American Society for Microbiology
Correspondence: gpier@channing.harvard.edu
Institutions: Charming Laboratory, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115-5804., Charming Laboratory, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School,Boston, Massachusetts 02115-5804.
- Article ID: 337
Newton GJ, Daniels C, Burrows LL, Kropinski AM, Clarke AJ, Lam JS "Three-component-mediated serotype conversion in Pseudomonas aeruginosa by bacteriophage D3" -
Molecular Microbiology 39(5) (2001) 1237-1247
Bacteriophage D3 is capable of lysogenizing Pseudomonas aeruginosa PAO1 (serotype O5), converting the O-antigen from O5 to O16 and O-acetylating the N-acetylfucosamine moiety. To investigate the mechanism of lysogenic conversion, a 3.6 kb fragment from the D3 genome was isolated capable of mediating serotypic conversion identical to the D3 lysogen strain (AK1380). The PAO1 transformants containing this 3.6 kb of D3 DNA exhibited identical lipopolysaccharide (LPS) banding patterns to serotype O16 in silver-stained SDS-PAGE gels and displayed reactivity to an antibody specific for O-acetyl groups. Further analysis led to the identification of three open reading frames (ORFs) required for serotype conversion: an α-polymerase inhibitor (iap); an O-acetylase (oac); and a β-polymerase (wzyβ). The α-polymerase inhibitor (Iap) is capable of inhibiting the assembly of the serotype-specific O5 B-band LPS and allows the phage-encoded β-polymerase (Wzyβ) to form new β-linked B-band LPS. The D3 phage also alters the LPS by the addition of O-acetyl groups to the FucNAc residue in the O-antigen repeat unit by the action of the D3 O-acetylase (Oac). These three components form a simple yet elegant system by which bacteriophage D3 is capable of altering the surface of P. aeruginosa PAO1
serotype, Pseudomonas, Pseudomonas aeruginosa, bacteriophage, conversion, serotype conversion
NCBI PubMed ID: 11251840Journal NLM ID: 8712028Publisher: Blackwell Publishing
Correspondence: jlam@uoguelph.ca
Institutions: Department of Microbiology, University of Guelph, Guelph, Ontario, N1G 2W1, Canada, The Centre for Infection and Biomaterials Research, Toronto General Hospital, Toronto, Ontario, M5G 2C4, Canada, Department of Microbiology and Immunology, Queen's University, Kingston, Ontario, K7L 3N6, Canada
Methods: PCR, SDS-PAGE, DNA techniques, Western blotting, quantification of O-acetylation
- Article ID: 1778
Knirel YA, Kochetkov NK "The structure of lipopolysaccharides of gram-negative bacteria. III. The structure of O-antigens: A review" -
Biochemistry (Moscow) 59(12) (1994) 1325-1383
This review summarizes data on the composition and structure of the O-antigens, the polysaccharide chains of the outer-membrane lipopolysaccharides (LPS) of Gram-negative bacteria defining the immunospecificity of these microbial cells. Special reference is given to some structural features of the O-antigens, such as the presence of unique monosaccharides and noncarbohydrate components, masked regularity, and the occurrence in one microorganism of LPS with structurally different polysaccharide chains. Antigenic relationships between microorganisms belonging to different taxonomic groups are discussed.
structure, O-antigen, chemical composition, bacterial lipopolysaccharides, Salmonella livingstone C1
NCBI PubMed ID: 7533007Journal NLM ID: 0376536Publisher: Nauka/Interperiodica
Institutions: Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia
- Article ID: 1842
Knirel YA, Paramonov NA, Vinogradov EV, Shashkov AS, Dmitriev BA, Kochetkov NK, Stanislavsky ES "Antigenic polysaccharides of bacteria. 20. Structure of the O-specific polysaccharide chain of P. aeruginosa O3(a),3d,3f (Lányi) lipopolysaccharide" -
Bioorganicheskaya Khimia = Bioorganic Chemistry [Russian] 12 (1986) 1649-1657
Mild acid degradation of lipopolysaccharide from Pseudomonas aeruginosa O(3a), 3d, 3f (Lányi classification) afforded O-specific polysaccharide containing N-acetyl-D-fucosamine, 2,3-diacetamido-2,3-dideoxy-D-mannuronic acid, 3-acetamidino-2-acetamido-2,3-dideoxy-L-guluronic and D-mannuronic acid as well as O-acetyl groups. On the basis of O-deacetylation, selective cleavage with anhydrous fluoride, chemical transformation of the oligosaccharides obtained (hydrolysis or reductive deamination of the acetamidino group into acetamido or ethylamino group, respectively) and analysis by 13C NMR spectroscopy, it was concluded that the polysaccharide is built up mainly by trisaccharide repeating units of types A and B in the ratio approximately 2:1: (Formula: see text). The units of both types most probably enter the same polymeric chain. If so, such a hybrid structure can be accounted for by incompleteness of epimerization at C5 of the acetamidino derivative of mannuronic acid at the polymer level in the course of biosynthesis of this polysaccharide.
NCBI PubMed ID: 2434103Journal NLM ID: 7804941Publisher: Moskva: Nauka
Institutions: N.D. Zelinsky Institute of Organic Chemistry, Academy of Sciences of the USSR, Moscow, Russia
Methods: 13C NMR, 1H NMR
- Article ID: 3207
Knirel YA, Bystrova OV, Kocharova NA, Zähringer U, Pier GB "Conserved and variable structural features of the Pseudomonas aeruginosa lipopolysaccharide" -
Journal of Endotoxin Research 12(6) (2006) 324-336
The review is devoted to recent progress in the structural elucidation of the lipopolysaccharide of the bacterium Pseudomonas aeruginosa, including O-antigen biological repeats, core oligosaccharide, and lipid A. Data on biosynthesis, genetics and serology of the lipopolysaccharide isolated from various P. aeruginosa O-serogroups are discussed in relation to the chemical structures.
O-antigen, Pseudomonas aeruginosa, lipid A, core oligosaccharide, lipopolysaccharide structure, serologic activity
NCBI PubMed ID: 17254386Publication DOI: 10.1179/096805106X118906Journal NLM ID: 9433350Publisher: Maney Publishing
Correspondence: knirel@ioc.ac.ru
Institutions: N.D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia, Research Center Borstel, Leibniz-Center for Medicine and Biosciences, Borstel, Germany, Channing Laboratory, Brigham and Women’s Hospital, Harvard Medical School, Boston, Massachusetts, USA
Methods: NMR, chemical methods, MS, serological methods, genetic methods
- Article ID: 3391
Kaluzny K, Abeyrathne PD, Lam JS "Coexistence of Two Distinct Versions of O-Antigen Polymerase, Wzy-Alpha and Wzy-Beta, in Pseudomonas aeruginosa Serogroup O2 and Their Contributions to Cell Surface Diversity" -
Journal of Bacteriology 189(11) (2007) 4141-4152
Assembly of B-band LPS in Pseudomonas aeruginosa follows a Wzy-dependent pathway, requiring the O-antigen polymerase Wzy, and other proteins. The peptide sequences of the wzyα product from strains of serotypes O2, O5, and O16 are identical; but the O-units in O5 are α-glycosidic-linked, while those in O2 and O16 are β-linked. We hypothesized that a derivative of the D3 bacteriophage wzyβ is present in the chromosomes of O2 and O16, and that this gene is responsible for the β-linkage. By a combination of PCR and primer walking, wzyβ of both serotypes have been amplified and cloned. Both are identical, but only share 87.42% sequence identity with their xenolog in D3. A chromosomal knockout mutant of O16 wzyβ was made and it produces rough LPS devoid of B-band O antigen. The cloned wzyβ is capable of complementing the O16 wzyβ mutant, as well as cross-complementing a wzyα knockout mutant. However, in the latter case, the restored O-antigen was β-linked. Using RT-PCR, we showed that wzyα was transcribed in O2 and O16 strains and was functional since both of these genes could complement the wzyα mutant of O5. With the co-existence of wzyα and wzyβ in O2 and O16 and the B-band O polysaccharides in these being β-linked, we hypothesized that iap, an inhibitor of alpha-polymerase gene, must be present in these serotypes. Indeed, through PCR, TOPO cloning and nucleotide sequencing results, we verified the presence of iap in both O2 and O16 serotypes
serotype, Pseudomonas aeruginosa, O-polysaccharide, serogroup, sequencing, O-antigen polymerase, rough LPS
NCBI PubMed ID: 17384183Journal NLM ID: 2985120RPublisher: American Society for Microbiology
Correspondence: jlam@uoguelph.ca
Institutions: University of Guelph, Department of Molecular and Cellular Biology, Guelph, Ontario, Canada N1G 2W1
Methods: serological methods, genetic methods
- Article ID: 4062
Lam JS, Taylor VL, Islam ST, Hao Y, Kocincova D "Genetic and Functional Diversity of Pseudomonas aeruginosa Lipopolysaccharide" -
Frontiers in Microbiology 2 (2011) 118
Lipopolysccharide (LPS) is an integral component of the Pseudomonas aeruginosa cell envelope, occupying the outer leaflet of the outer membrane in this Gram-negative opportunistic pathogen. It is important for bacterium-host interactions and has been shown to be a major virulence factor for this organism. Structurally, P. aeruginosa LPS is composed of three domains, namely, lipid A, core oligosaccharide, and the distal O antigen (O-Ag). Most P. aeruginosa strains produce two distinct forms of O-Ag, one a homopolymer of D-rhamnose that is a common polysaccharide antigen (CPA, formerly termed A band), and the other a heteropolymer of three to five distinct (and often unique dideoxy) sugars in its repeat units, known as O-specific antigen (OSA, formerly termed B band). Compositional differences in the O units among the OSA from different strains form the basis of the International Antigenic Typing Scheme for classification via serotyping of different strains of P. aeruginosa. The focus of this review is to provide state-of-the-art knowledge on the genetic and resultant functional diversity of LPS produced by P. aeruginosa. The underlying factors contributing to this diversity will be thoroughly discussed and presented in the context of its contributions to host-pathogen interactions and the control/prevention of infection.
Lipopolysaccharide, biosynthesis, virulence, serotyping, bacteriophage, motility, seroconversion, nucleotide sugars
NCBI PubMed ID: 21687428Publication DOI: 10.3389/fmicb.2011.00118Journal NLM ID: 101548977Publisher: Lausanne: Frontiers Research Foundation
Correspondence: jlam@uoguelph.ca
Institutions: Department of Molecular and Cellular Biology, University of Guelph Guelph, ON, Canada
- Article ID: 4684
Islam ST, Lam JS "Synthesis of bacterial polysaccharides via the Wzx/Wzy-dependent pathway" -
Canadian Journal of Microbiology 60(11) (2014) 697-716
The surfaces of bacteria mediate a multitude of functions in the environment and in an infected host, including adhesion to both biotic and abiotic substrata, motility, immune system interaction and (or) activation, biofilm formation, and cell-cell communication, with many of these features directly influenced by cell-surface glycans. In both Gram-negative and Gram-positive bacteria, the majority of cell-surface polysaccharides are produced via the Wzx/Wzy-dependent assembly pathway; these glycans include heteropolymeric O-antigen, enterobacterial common antigen, exopolysaccharide, spore coat, and capsule in diverse bacteria. The key components of this assembly pathway are the integral inner membrane Wzx flippase, Wzy polymerase, and Wzz chain-length regulator proteins, which until recently have resisted detailed structural and functional characterization. In this review, we have provided a comprehensive synthesis of the latest structural and mechanistic data for each protein, as well as an examination of substrate specificity for each assembly step and complex formation between the constituent proteins. To complement the unprecedented explosion of genomic-sequencing data for bacteria, we have also highlighted both classical and state-of-the-art methods by which encoded Wzx, Wzy, and Wzz proteins can be reliably identified and annotated, using the model Gram-negative bacterium Pseudomonas aeruginosa as an example data set. Lastly, we outline future avenues of research, with the aim of stimulating researchers to take the next steps in investigating the function of, and interplay between, the constituents of this widespread assembly scheme.
Membrane Proteins, lipopolysaccharide (LPS), Wzx flippase, Wzy polymerase, Wzz polysaccharide copolymerase
NCBI PubMed ID: 25358682Publication DOI: 10.1139/cjm-2014-0595Journal NLM ID: 0372707Publisher: National Research Council of Canada
Correspondence: sislam@imm.cnrs.fr; jlam@uoguelph.ca
Institutions: Department of Molecular and Cellular Biology, University of Guelph, Guelph, ON N1G 2W1, Canada
Methods: SDS-PAGE, genetic methods, cloning
- Article ID: 4932
Taylor VL, Hoage JF, Thrane SW, Huszczynski SM, Jelsbak L, Lam JS "A Bacteriophage-Acquired O-Antigen Polymerase (Wzyb) from P. aeruginosa Serotype O16 Performs a Varied Mechanism Compared to Its Cognate Wzya" -
Frontiers in Microbiology 7 (2016) 393
Pseudomonas aeruginosa is a Gram-negative bacterium that produces highly varied lipopolysaccharide (LPS) structures. The O antigen (O-Ag) in the LPS is synthesized through the Wzx/Wzy-dependent pathway where lipid-linked O-Ag repeats are polymerized by Wzy. Horizontal-gene transfer has been associated with O-Ag diversity. The O-Ag present on the surface of serotypes O5 and O16, differ in the intra-molecular bonds, alpha and beta, respectively; the latter arose from the action of three genes in a serotype converting unit acquired from bacteriophage D3, including a beta-polymerase (Wzyβ). To further our understanding of O-polymerases, the inner membrane (IM) topology of Wzyβ was determined using a dual phoA-lacZα reporter system wherein random 3' gene truncations were localized to specific loci with respect to the IM by normalized reporter activities as determined through the ratio of alkaline phosphatase activity to β-galactosidase activity. The topology of Wzyβ developed through this approach was shown to contain two predominant periplasmic loops, PL3 (containing an RX10G motif) and PL4 (having an O-Ag ligase superfamily motif), associated with inverting glycosyltransferase reaction. Through site-directed mutagenesis and complementation assays, residues Arg(254), Arg(270), Arg(272), and His(300) were found to be essential for Wzyβ function. Additionally, like-charge substitutions, R254K and R270K, could not complement the wzyβ knockout, highlighting the essential guanidium side group of Arg residues. The O-Ag ligase domain is conserved among heterologous Wzy proteins that produce β-linked O-Ag repeat units. Taking advantage of the recently obtained whole-genome sequence of serotype O16 a candidate promoter was identified. Wzyβ under its native promoter was integrated in the PAO1 genome, which resulted in simultaneous production of α- and β-linked O-Ag. These observations established that members of Wzy-like family consistently exhibit a dual-periplasmic loops topology, and identifies motifs that are plausible to be involved in enzymatic activities. Based on these results, the phage-derived Wzyβ utilizes a different reaction mechanism in the P. aeruginosa host to avoid self-inhibition during serotype conversion.
Lipopolysaccharide, serotype, Pseudomonas aeruginosa, glycosyltransferase, bacteriophage, O-antigen biosynthesis, polymerase
NCBI PubMed ID: 27065964Publication DOI: 10.3389/fmicb.2016.00393Journal NLM ID: 101548977Publisher: Lausanne: Frontiers Research Foundation
Correspondence: jlam@uoguelph.ca
Institutions: Department of Molecular and Cellular Biology, University of Guelph Guelph, ON, Canada, Department of Systems Biology, Technical University of Denmark Kongens Lyngby, Denmark
Methods: PCR, SDS-PAGE, Western blotting, genetic methods, enzymatic assay
- Article ID: 6301
Qin CJ, Ding MR, Tian GZ, Zou XP, Fu JJ, Hu J, Yin J "Chemical approaches towards installation of rare functional groups in bacterial surface glycans" -
Chinese Journal of Natural Medicines = Zhongguo Tianran Yaowu 20(6) (2022) 401-420
Bacterial surface glycans perform a diverse and important set of biological roles, and have been widely used in the treatment of bacterial infectious diseases. The majority of bacterial surface glycans are decorated with diverse rare functional groups, including amido, acetamidino, carboxamido and pyruvate groups. These functional groups are thought to be important constituents for the biological activities of glycans. Chemical synthesis of glycans bearing these functional groups or their variants is essential for the investigation of structure-activity relationships by a medicinal chemistry approach. To date, a broad choice of synthetic methods is available for targeting the different rare functional groups in bacterial surface glycans. This article reviews the structures of naturally occurring rare functional groups in bacterial surface glycans, and the chemical methods used for installation of these groups.
chemical synthesis, acetamidino group, amido group, bacterial surface glycan, carboxamido group, pyruvyl ketal
NCBI PubMed ID: 35750381Publication DOI: 10.1016/S1875-5364(22)60177-8Journal NLM ID: 101504416Publisher: Beijing: Science Press; Elsevier
Correspondence: J. Yin
Institutions: Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, China, Wuxi School of Medicine, Jiangnan University, Wuxi, China
Expand this compound
Collapse this compound
11. Compound ID: 968
|
-4)-a-L-GulpNAc3NAmA-(1-4)-b-D-ManpNAc3NAcA-(1-3)-a-D-FucpNAc-(1- |
Show graphically |
Structure type: polymer chemical repeating unit
Compound class: O-polysaccharide, O-antigen
The structure is contained in the following publication(s):
- Article ID: 254
Hatano K, Pier GB "Complex serology and immune response of mice to variant high-molecular-weight O polysaccharide isolated from Pseudomonas aeruginosa serogroup O2 strains" -
Infection and Immunity 66(8) (1998) 3719-3726
The O antigen of the Pseudomonas aeruginosa lipopolysaccharide is the optimal target for protective antibodies, but the unusual and complex nature of their sugar substituents has made it difficult to define the range of these structures needed in an effective vaccine. Most clinical isolates of P. aeruginosa can be classified into 10 O-antigen serogroups, but slight chemical differences among O polysaccharides within a serogroup give rise to subtype epitopes. These epitopes could impact the reactivity of O-antigen-specific antibodies, as well as the susceptibility of a target strain to protective, opsonic antibodies. To define parameters of serogroup and subtype-epitope immunogenicity, antigenicity, and surface expression on P. aeruginosa cells, we prepared high-molecular-weight O-polysaccharide vaccines from strains of P. aeruginosa serogroup O2, for which eight structurally variant O antigens expressing six defined subtype epitopes (O2a to O2f) have been identified. A complex pattern of immune responses to these antigens was observed following vaccination of mice. The high-molecular-weight O polysaccharides were generally more immunogenic at low doses (1 and 10 microg) than at a high dose (50 microg) and usually elicited antibodies that opsonized the homologous strain for phagocytic killing. Some of the individual polysaccharides elicited cross-opsonic antibodies to a variable number of strains that express all of the defined serogroup O2 subtype epitopes. Combination into one vaccine of two antigens that individually elicited cross-reactive opsonic antibodies to most members of the O2 serogroup inhibited, instead of enhanced, the production of antibodies broadly reactive with most serogroup O2 subtype strains. Thus, immune responses to P. aeruginosa O antigens may be restricted to a limited range of epitopes on structurally complex O antigens, and combining multiple related antigens into a single vaccine formulation may inhibit the production of those antibodies best able to protect against most P. aeruginosa strains within a given O-antigen serogroup.
Lipopolysaccharide, Pseudomonas, Pseudomonas aeruginosa, O-polysaccharide, serology, serogroup, immune response
NCBI PubMed ID: 9673254Journal NLM ID: 0246127Publisher: American Society for Microbiology
Correspondence: gpier@channing.harvard.edu
Institutions: Charming Laboratory, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115-5804., Charming Laboratory, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School,Boston, Massachusetts 02115-5804.
- Article ID: 337
Newton GJ, Daniels C, Burrows LL, Kropinski AM, Clarke AJ, Lam JS "Three-component-mediated serotype conversion in Pseudomonas aeruginosa by bacteriophage D3" -
Molecular Microbiology 39(5) (2001) 1237-1247
Bacteriophage D3 is capable of lysogenizing Pseudomonas aeruginosa PAO1 (serotype O5), converting the O-antigen from O5 to O16 and O-acetylating the N-acetylfucosamine moiety. To investigate the mechanism of lysogenic conversion, a 3.6 kb fragment from the D3 genome was isolated capable of mediating serotypic conversion identical to the D3 lysogen strain (AK1380). The PAO1 transformants containing this 3.6 kb of D3 DNA exhibited identical lipopolysaccharide (LPS) banding patterns to serotype O16 in silver-stained SDS-PAGE gels and displayed reactivity to an antibody specific for O-acetyl groups. Further analysis led to the identification of three open reading frames (ORFs) required for serotype conversion: an α-polymerase inhibitor (iap); an O-acetylase (oac); and a β-polymerase (wzyβ). The α-polymerase inhibitor (Iap) is capable of inhibiting the assembly of the serotype-specific O5 B-band LPS and allows the phage-encoded β-polymerase (Wzyβ) to form new β-linked B-band LPS. The D3 phage also alters the LPS by the addition of O-acetyl groups to the FucNAc residue in the O-antigen repeat unit by the action of the D3 O-acetylase (Oac). These three components form a simple yet elegant system by which bacteriophage D3 is capable of altering the surface of P. aeruginosa PAO1
serotype, Pseudomonas, Pseudomonas aeruginosa, bacteriophage, conversion, serotype conversion
NCBI PubMed ID: 11251840Journal NLM ID: 8712028Publisher: Blackwell Publishing
Correspondence: jlam@uoguelph.ca
Institutions: Department of Microbiology, University of Guelph, Guelph, Ontario, N1G 2W1, Canada, The Centre for Infection and Biomaterials Research, Toronto General Hospital, Toronto, Ontario, M5G 2C4, Canada, Department of Microbiology and Immunology, Queen's University, Kingston, Ontario, K7L 3N6, Canada
Methods: PCR, SDS-PAGE, DNA techniques, Western blotting, quantification of O-acetylation
- Article ID: 1778
Knirel YA, Kochetkov NK "The structure of lipopolysaccharides of gram-negative bacteria. III. The structure of O-antigens: A review" -
Biochemistry (Moscow) 59(12) (1994) 1325-1383
This review summarizes data on the composition and structure of the O-antigens, the polysaccharide chains of the outer-membrane lipopolysaccharides (LPS) of Gram-negative bacteria defining the immunospecificity of these microbial cells. Special reference is given to some structural features of the O-antigens, such as the presence of unique monosaccharides and noncarbohydrate components, masked regularity, and the occurrence in one microorganism of LPS with structurally different polysaccharide chains. Antigenic relationships between microorganisms belonging to different taxonomic groups are discussed.
structure, O-antigen, chemical composition, bacterial lipopolysaccharides, Salmonella livingstone C1
NCBI PubMed ID: 7533007Journal NLM ID: 0376536Publisher: Nauka/Interperiodica
Institutions: Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia
- Article ID: 1832
Knirel YA, Paramonov NA, Vinogradov EV, Shashkov AS, Kochetkov NK "Identification of 3-acetamidino-2-acetamido-2,3-dideoxy-L-guluronic acid in Pseudomonas aeruginosa immunotype 7 lipopolysaccharide" -
Bioorganicheskaya Khimia = Bioorganic Chemistry [Russian] 12 (1986) 992-994
O-Specific polysaccharide chain of Pseudomonas aeruginosa immunotype 7 lipopolysaccharide is composed of 3-acetamidino-2-acetamido-2,3-dideoxy-L-guluronic acid (GulNAcAmA), 2,3-diacetamido-2,3-dideoxy-D-mannuronic acid (ManN2Ac2A), and N-acetyl-D-fucosamine (FucNAc). On solvolysis with anhydrous hydrogen fluoride, the polysaccharide afforded a trisaccharide containing all its components. Borohydride reduction of the trisaccharide in boric acid solution resulted in conversion of reducing fucosamine into fucosaminitol, whereas in water the reduction was accompanied by reductive deamination of acetamidino function into ethylamino group. On hydrolysis with aqueous triethylamine, acetamidino group gave acetamido group. Analysis of the trisaccharides thus obtained by 1H NMR spectroscopy (including nuclear Overhauser effect), 13C NMR spectroscopy, and fast-atom bombardment mass spectrometry allowed the determination of the structure of the unusual uronic acid derivative and the following structure of the polysaccharide repeating unit: -4)-α-L-GulNAcAmA-(1-4)-β-D-ManN2Ac2A-(1-3)-α-D-FucNAc-(1-.
NCBI PubMed ID: 2429670Journal NLM ID: 7804941Publisher: Moskva: Nauka
Institutions: N.D. Zelinsky Institute of Organic Chemistry, Academy of Sciences of the USSR, Moscow, Russia
Methods: 13C NMR
- Article ID: 1842
Knirel YA, Paramonov NA, Vinogradov EV, Shashkov AS, Dmitriev BA, Kochetkov NK, Stanislavsky ES "Antigenic polysaccharides of bacteria. 20. Structure of the O-specific polysaccharide chain of P. aeruginosa O3(a),3d,3f (Lányi) lipopolysaccharide" -
Bioorganicheskaya Khimia = Bioorganic Chemistry [Russian] 12 (1986) 1649-1657
Mild acid degradation of lipopolysaccharide from Pseudomonas aeruginosa O(3a), 3d, 3f (Lányi classification) afforded O-specific polysaccharide containing N-acetyl-D-fucosamine, 2,3-diacetamido-2,3-dideoxy-D-mannuronic acid, 3-acetamidino-2-acetamido-2,3-dideoxy-L-guluronic and D-mannuronic acid as well as O-acetyl groups. On the basis of O-deacetylation, selective cleavage with anhydrous fluoride, chemical transformation of the oligosaccharides obtained (hydrolysis or reductive deamination of the acetamidino group into acetamido or ethylamino group, respectively) and analysis by 13C NMR spectroscopy, it was concluded that the polysaccharide is built up mainly by trisaccharide repeating units of types A and B in the ratio approximately 2:1: (Formula: see text). The units of both types most probably enter the same polymeric chain. If so, such a hybrid structure can be accounted for by incompleteness of epimerization at C5 of the acetamidino derivative of mannuronic acid at the polymer level in the course of biosynthesis of this polysaccharide.
NCBI PubMed ID: 2434103Journal NLM ID: 7804941Publisher: Moskva: Nauka
Institutions: N.D. Zelinsky Institute of Organic Chemistry, Academy of Sciences of the USSR, Moscow, Russia
Methods: 13C NMR, 1H NMR
- Article ID: 1843
Elkin YN, Knirel YA, Vinogradov EV, Paramonov NA, Troshkov ML, Aminev "Fast-atom-bombardment mass-spectra of aminooligosaccharides" -
Bioorganicheskaya Khimia = Bioorganic Chemistry [Russian] 12 (1986) 1658-1661
Journal NLM ID: 7804941Publisher: Moskva: Nauka
Institutions: N.D. Zelinsky Institute of Organic Chemistry, Academy of Sciences of the USSR, Moscow, Russia
Methods: FAB-MS
- Article ID: 1844
Knirel YA, Kocharova NA, Vinogradov EV, Paramonov NA, Dmitriev BA, Kochetkov NK, Stanislavsky ES, Kholodkova EV "Antigenic polysaccharides of bacteria. 21. Structure of O-specific polysaccharide chains and serological specificity of lipopolysaccharides of seven Pseudomonas aeruginosa immunotypes" -
Bioorganicheskaya Khimia = Bioorganic Chemistry [Russian] 13 (1987) 88-96
On mild acid degradation on lipopolysaccharides of seven Pseudomonas aeruginosa immunotypes, O-specific polysaccharides were obtained and their structures established. A peculiar feature of the polysaccharides is the presence of various, mostly acidic, mono- and diaminosugars, many of which have not previously been found in nature. The absence of serological cross-reactions (inhibition of passive haemagglutination) between lipopolysaccharides of seven immunotypes correlates with the absence of any common oligosaccharide fragments in their O-specific chains. The data obtained revealed structural and serological interrelations between O-antigens of seven immunotypes and P. aeruginosa O-serotypes, and showed that immunotypes 1 and 7 should be included into the serological classification scheme as individual O-serotypes.
NCBI PubMed ID: 2436629Journal NLM ID: 7804941Publisher: Moskva: Nauka
Institutions: N.D. Zelinsky Institute of Organic Chemistry, Academy of Sciences of the USSR, Moscow, Russia
- Article ID: 2409
Knirel YA, Paramonov NA, Vinogradov EV, Shashkov AS, Dmitriev BA, Kochetkov NK, Kholodkova EV, Stanislavsky ES "Somatic antigens of Pseudomonas aeruginosa. The structure of O-specific polysaccharide chains of lipopolysaccharides of P. aeruginosa O3 (Lányi), O25 (Wokatsch) and Fisher immunotypes 3 and 7" -
European Journal of Biochemistry 167 (1987) 549-561
O-specific polysaccharides, obtained on mild acid degradation of lipopolysacchrides of the serologically related strains Pseudomonas aeruginosa O3 (Lányi classification), O25 (Wokatsch classification) and immunotypes 3 and 7 (Fisher classification), are built up of trisaccharide repeating units involving 2-acetamido-2,6-dideoxy-D-galactose (N-acetyl-D-fucosamine), 2,3-diacetamido-2,3-dideoxy-D-mannuronic acid or 2,3-diacetamido-2,3-dideoxy-L-guluronic acid and 3-acetamidino-2-acetamido-2,3-dideoxy-D-mannuronic acid or 3-acetamidino-2-acetamido-2,3-dideoxy-L-guluronic acid. Lányi O3(a),3d,3f and Wokatsch O25 polysaccharides contain also O-acetyl groups. On the basis of solvolysis with anhydrous hydrogen fluoride, resulting in trisaccharide fragments with N-acetylfucosamine residue at the reducing terminus, chemical modifications of the acetamidino group (alkaline hydrolysis to the acetamido group or reductive deamination to the ethylamino group), as well as analysis by 1H-NMR (including nuclear Overhauser effect experiments) and 13C-NMR spectroscopy, and fast-atom bombardment mass spectrometry, it was concluded that the repeating units of the polysaccharides have the following structures: (Formula: see text) where HexNAcAmA = α-L-GulNAcAmA (approximately 70%) or β-D-ManNacAMA (approximately 30%). Lányi O3(a),3d,3f polysaccharide involves two types of repeating units, which differ from each other only in the configuration at C-5 of the 3-acetamidino-2-acetamido-2,3-dideoxyuronic acid residue. Lányi O3(a),3c,O3a,3d,3e and Fisher immunotypes 3 and 7 polysaccharides contain, together with the major repeating units shown above, a small proportion of units in which the derivative of α-L-guluronic acid is replaced by the corresponding β-D-manno isomer. The data obtained provide the opportunity to substantiate the serological interrelations between these strains of P. aeruginosa by the presence in the O-specific polysaccharides of common monosaccharides or disaccharide fragments. The distinctions between them stem from the presence or absence of the O-acetyl group, a different configuration of the glycosidic linkage of the N-acetylfucosamine residue and/or a different configuration at C-5 of one or both derivatives of diaminouronic acids.
NCBI PubMed ID: 3115777Journal NLM ID: 0107600Publisher: Oxford, UK: Blackwell Science Ltd. on behalf of the Federation of European Biochemical Societies
Institutions: N.D. Zelinsky Institute of Organic Chemistry, Academy of Sciences of the USSR, Moscow, Russia
Methods: 13C NMR, 1H NMR, FAB-MS
- Article ID: 3207
Knirel YA, Bystrova OV, Kocharova NA, Zähringer U, Pier GB "Conserved and variable structural features of the Pseudomonas aeruginosa lipopolysaccharide" -
Journal of Endotoxin Research 12(6) (2006) 324-336
The review is devoted to recent progress in the structural elucidation of the lipopolysaccharide of the bacterium Pseudomonas aeruginosa, including O-antigen biological repeats, core oligosaccharide, and lipid A. Data on biosynthesis, genetics and serology of the lipopolysaccharide isolated from various P. aeruginosa O-serogroups are discussed in relation to the chemical structures.
O-antigen, Pseudomonas aeruginosa, lipid A, core oligosaccharide, lipopolysaccharide structure, serologic activity
NCBI PubMed ID: 17254386Publication DOI: 10.1179/096805106X118906Journal NLM ID: 9433350Publisher: Maney Publishing
Correspondence: knirel@ioc.ac.ru
Institutions: N.D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia, Research Center Borstel, Leibniz-Center for Medicine and Biosciences, Borstel, Germany, Channing Laboratory, Brigham and Women’s Hospital, Harvard Medical School, Boston, Massachusetts, USA
Methods: NMR, chemical methods, MS, serological methods, genetic methods
- Article ID: 3391
Kaluzny K, Abeyrathne PD, Lam JS "Coexistence of Two Distinct Versions of O-Antigen Polymerase, Wzy-Alpha and Wzy-Beta, in Pseudomonas aeruginosa Serogroup O2 and Their Contributions to Cell Surface Diversity" -
Journal of Bacteriology 189(11) (2007) 4141-4152
Assembly of B-band LPS in Pseudomonas aeruginosa follows a Wzy-dependent pathway, requiring the O-antigen polymerase Wzy, and other proteins. The peptide sequences of the wzyα product from strains of serotypes O2, O5, and O16 are identical; but the O-units in O5 are α-glycosidic-linked, while those in O2 and O16 are β-linked. We hypothesized that a derivative of the D3 bacteriophage wzyβ is present in the chromosomes of O2 and O16, and that this gene is responsible for the β-linkage. By a combination of PCR and primer walking, wzyβ of both serotypes have been amplified and cloned. Both are identical, but only share 87.42% sequence identity with their xenolog in D3. A chromosomal knockout mutant of O16 wzyβ was made and it produces rough LPS devoid of B-band O antigen. The cloned wzyβ is capable of complementing the O16 wzyβ mutant, as well as cross-complementing a wzyα knockout mutant. However, in the latter case, the restored O-antigen was β-linked. Using RT-PCR, we showed that wzyα was transcribed in O2 and O16 strains and was functional since both of these genes could complement the wzyα mutant of O5. With the co-existence of wzyα and wzyβ in O2 and O16 and the B-band O polysaccharides in these being β-linked, we hypothesized that iap, an inhibitor of alpha-polymerase gene, must be present in these serotypes. Indeed, through PCR, TOPO cloning and nucleotide sequencing results, we verified the presence of iap in both O2 and O16 serotypes
serotype, Pseudomonas aeruginosa, O-polysaccharide, serogroup, sequencing, O-antigen polymerase, rough LPS
NCBI PubMed ID: 17384183Journal NLM ID: 2985120RPublisher: American Society for Microbiology
Correspondence: jlam@uoguelph.ca
Institutions: University of Guelph, Department of Molecular and Cellular Biology, Guelph, Ontario, Canada N1G 2W1
Methods: serological methods, genetic methods
- Article ID: 4062
Lam JS, Taylor VL, Islam ST, Hao Y, Kocincova D "Genetic and Functional Diversity of Pseudomonas aeruginosa Lipopolysaccharide" -
Frontiers in Microbiology 2 (2011) 118
Lipopolysccharide (LPS) is an integral component of the Pseudomonas aeruginosa cell envelope, occupying the outer leaflet of the outer membrane in this Gram-negative opportunistic pathogen. It is important for bacterium-host interactions and has been shown to be a major virulence factor for this organism. Structurally, P. aeruginosa LPS is composed of three domains, namely, lipid A, core oligosaccharide, and the distal O antigen (O-Ag). Most P. aeruginosa strains produce two distinct forms of O-Ag, one a homopolymer of D-rhamnose that is a common polysaccharide antigen (CPA, formerly termed A band), and the other a heteropolymer of three to five distinct (and often unique dideoxy) sugars in its repeat units, known as O-specific antigen (OSA, formerly termed B band). Compositional differences in the O units among the OSA from different strains form the basis of the International Antigenic Typing Scheme for classification via serotyping of different strains of P. aeruginosa. The focus of this review is to provide state-of-the-art knowledge on the genetic and resultant functional diversity of LPS produced by P. aeruginosa. The underlying factors contributing to this diversity will be thoroughly discussed and presented in the context of its contributions to host-pathogen interactions and the control/prevention of infection.
Lipopolysaccharide, biosynthesis, virulence, serotyping, bacteriophage, motility, seroconversion, nucleotide sugars
NCBI PubMed ID: 21687428Publication DOI: 10.3389/fmicb.2011.00118Journal NLM ID: 101548977Publisher: Lausanne: Frontiers Research Foundation
Correspondence: jlam@uoguelph.ca
Institutions: Department of Molecular and Cellular Biology, University of Guelph Guelph, ON, Canada
- Article ID: 4684
Islam ST, Lam JS "Synthesis of bacterial polysaccharides via the Wzx/Wzy-dependent pathway" -
Canadian Journal of Microbiology 60(11) (2014) 697-716
The surfaces of bacteria mediate a multitude of functions in the environment and in an infected host, including adhesion to both biotic and abiotic substrata, motility, immune system interaction and (or) activation, biofilm formation, and cell-cell communication, with many of these features directly influenced by cell-surface glycans. In both Gram-negative and Gram-positive bacteria, the majority of cell-surface polysaccharides are produced via the Wzx/Wzy-dependent assembly pathway; these glycans include heteropolymeric O-antigen, enterobacterial common antigen, exopolysaccharide, spore coat, and capsule in diverse bacteria. The key components of this assembly pathway are the integral inner membrane Wzx flippase, Wzy polymerase, and Wzz chain-length regulator proteins, which until recently have resisted detailed structural and functional characterization. In this review, we have provided a comprehensive synthesis of the latest structural and mechanistic data for each protein, as well as an examination of substrate specificity for each assembly step and complex formation between the constituent proteins. To complement the unprecedented explosion of genomic-sequencing data for bacteria, we have also highlighted both classical and state-of-the-art methods by which encoded Wzx, Wzy, and Wzz proteins can be reliably identified and annotated, using the model Gram-negative bacterium Pseudomonas aeruginosa as an example data set. Lastly, we outline future avenues of research, with the aim of stimulating researchers to take the next steps in investigating the function of, and interplay between, the constituents of this widespread assembly scheme.
Membrane Proteins, lipopolysaccharide (LPS), Wzx flippase, Wzy polymerase, Wzz polysaccharide copolymerase
NCBI PubMed ID: 25358682Publication DOI: 10.1139/cjm-2014-0595Journal NLM ID: 0372707Publisher: National Research Council of Canada
Correspondence: sislam@imm.cnrs.fr; jlam@uoguelph.ca
Institutions: Department of Molecular and Cellular Biology, University of Guelph, Guelph, ON N1G 2W1, Canada
Methods: SDS-PAGE, genetic methods, cloning
- Article ID: 4932
Taylor VL, Hoage JF, Thrane SW, Huszczynski SM, Jelsbak L, Lam JS "A Bacteriophage-Acquired O-Antigen Polymerase (Wzyb) from P. aeruginosa Serotype O16 Performs a Varied Mechanism Compared to Its Cognate Wzya" -
Frontiers in Microbiology 7 (2016) 393
Pseudomonas aeruginosa is a Gram-negative bacterium that produces highly varied lipopolysaccharide (LPS) structures. The O antigen (O-Ag) in the LPS is synthesized through the Wzx/Wzy-dependent pathway where lipid-linked O-Ag repeats are polymerized by Wzy. Horizontal-gene transfer has been associated with O-Ag diversity. The O-Ag present on the surface of serotypes O5 and O16, differ in the intra-molecular bonds, alpha and beta, respectively; the latter arose from the action of three genes in a serotype converting unit acquired from bacteriophage D3, including a beta-polymerase (Wzyβ). To further our understanding of O-polymerases, the inner membrane (IM) topology of Wzyβ was determined using a dual phoA-lacZα reporter system wherein random 3' gene truncations were localized to specific loci with respect to the IM by normalized reporter activities as determined through the ratio of alkaline phosphatase activity to β-galactosidase activity. The topology of Wzyβ developed through this approach was shown to contain two predominant periplasmic loops, PL3 (containing an RX10G motif) and PL4 (having an O-Ag ligase superfamily motif), associated with inverting glycosyltransferase reaction. Through site-directed mutagenesis and complementation assays, residues Arg(254), Arg(270), Arg(272), and His(300) were found to be essential for Wzyβ function. Additionally, like-charge substitutions, R254K and R270K, could not complement the wzyβ knockout, highlighting the essential guanidium side group of Arg residues. The O-Ag ligase domain is conserved among heterologous Wzy proteins that produce β-linked O-Ag repeat units. Taking advantage of the recently obtained whole-genome sequence of serotype O16 a candidate promoter was identified. Wzyβ under its native promoter was integrated in the PAO1 genome, which resulted in simultaneous production of α- and β-linked O-Ag. These observations established that members of Wzy-like family consistently exhibit a dual-periplasmic loops topology, and identifies motifs that are plausible to be involved in enzymatic activities. Based on these results, the phage-derived Wzyβ utilizes a different reaction mechanism in the P. aeruginosa host to avoid self-inhibition during serotype conversion.
Lipopolysaccharide, serotype, Pseudomonas aeruginosa, glycosyltransferase, bacteriophage, O-antigen biosynthesis, polymerase
NCBI PubMed ID: 27065964Publication DOI: 10.3389/fmicb.2016.00393Journal NLM ID: 101548977Publisher: Lausanne: Frontiers Research Foundation
Correspondence: jlam@uoguelph.ca
Institutions: Department of Molecular and Cellular Biology, University of Guelph Guelph, ON, Canada, Department of Systems Biology, Technical University of Denmark Kongens Lyngby, Denmark
Methods: PCR, SDS-PAGE, Western blotting, genetic methods, enzymatic assay
- Article ID: 6301
Qin CJ, Ding MR, Tian GZ, Zou XP, Fu JJ, Hu J, Yin J "Chemical approaches towards installation of rare functional groups in bacterial surface glycans" -
Chinese Journal of Natural Medicines = Zhongguo Tianran Yaowu 20(6) (2022) 401-420
Bacterial surface glycans perform a diverse and important set of biological roles, and have been widely used in the treatment of bacterial infectious diseases. The majority of bacterial surface glycans are decorated with diverse rare functional groups, including amido, acetamidino, carboxamido and pyruvate groups. These functional groups are thought to be important constituents for the biological activities of glycans. Chemical synthesis of glycans bearing these functional groups or their variants is essential for the investigation of structure-activity relationships by a medicinal chemistry approach. To date, a broad choice of synthetic methods is available for targeting the different rare functional groups in bacterial surface glycans. This article reviews the structures of naturally occurring rare functional groups in bacterial surface glycans, and the chemical methods used for installation of these groups.
chemical synthesis, acetamidino group, amido group, bacterial surface glycan, carboxamido group, pyruvyl ketal
NCBI PubMed ID: 35750381Publication DOI: 10.1016/S1875-5364(22)60177-8Journal NLM ID: 101504416Publisher: Beijing: Science Press; Elsevier
Correspondence: J. Yin
Institutions: Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, China, Wuxi School of Medicine, Jiangnan University, Wuxi, China
Expand this compound
Collapse this compound
12. Compound ID: 2350
|
L-Orn-(5-6)-+ D-Asp-(1-4)-+
| |
-4)-b-D-GlcpNAc3NAcA-(1-4)-b-D-ManpNAc3NA-(1-3)-b-D-GlcpNAc-(1-3)-a-D-Fucp4N-(1- |
Show graphically |
Structure type: polymer chemical repeating unit
Trivial name: polysaccharide part of a glycoconjugate, glycan repeating unit of the glycoconjugate
Contained glycoepitopes: IEDB_135813,IEDB_137340,IEDB_141807,IEDB_151531
The structure is contained in the following publication(s):
- Article ID: 806
Hashimoto M, Asai Y, Jinno T, Adachi S, Kusumoto S "Structural elucidation of polysaccharide part of glycoconjugate from Treponema medium ATCC700293" -
European Journal of Biochemistry 270 (2003) 2671-2679
Glycoconjugates are distributed on the cell surfaces of some small-sized treponemes and have been reported to be completely different from lipopolysaccharides. We separated a glycoconjugate fraction from Treponema medium ATCC700293, a medium-sized oral spirochete, to assess its immunobiological activities and elucidate the chemical structure of its polysaccharide part using phenol/water extraction, hydrophobic chromatography, and gel .ltration. The glycoconjugate showed negligible or weak endotoxic and immunobiological properties. The chemical structure of the polysaccharide part was shown by two-dimensional NMR and MALDI-TOF-MS to be a tetrasaccharide backbone with two amino acids: -4)[Ac(1-3),Ac(1-2)]bDGlcpN3NA(1-4)[xLOrn(6-6)Ac(1-2)]bDManpN3NA(1-3)[Ac(1-2)]bDGlcpN(1-3)[xDAsp?(1-4)]aDFucp4N(1-.
structural, polysaccharide, aspartic acid, MALDI-TOF MS, ornithine, glycoconjugate, medium, Treponema
NCBI PubMed ID: 12787034Journal NLM ID: 0107600Publisher: Oxford, UK: Blackwell Science Ltd. on behalf of the Federation of European Biochemical Societies
Correspondence: tomo527@dent.asahi-u.ac.jp
Institutions: Department of Oral Microbiology, Asahi University School of Dentistry, Gifu, Japan, Graduate School of Science, Osaka University, Osaka, Japan
Methods: NMR-2D, ESI-MS, MALDI-TOF MS
- Article ID: 3697
Harvey DJ "Analysis of carbohydrates and glycoconjugates by matrix-assisted laser desorption/ionization mass spectrometry: An update for 2003-2004" -
Mass Spectrometry Reviews 28(2) (2009) 273-361
This review is the third update of the original review, published in 1999, on the application of matrix-assisted laser desorption/ionization (MALDI) mass spectrometry to the analysis of carbohydrates and glycoconjugates and brings the topic to the end of 2004. Both fundamental studies and applications are covered. The main topics include methodological developments, matrices, fragmentation of carbohydrates and applications to large polymeric carbohydrates from plants, glycans from glycoproteins and those from various glycolipids. Other topics include the use of MALDI MS to study enzymes related to carbohydrate biosynthesis and degradation, its use in industrial processes, particularly biopharmaceuticals and its use to monitor products of chemical synthesis where glycodendrimers and carbohydrate-protein complexes are highlighted
carbohydrates, glycosyltransferases, fragmentation, MALDI, glycolipids, glycoproteins, biopharmaceuticals, glycosidases, time-of-flight
NCBI PubMed ID: 18825656Publication DOI: 10.1002/mas.2019Journal NLM ID: 8219702Publisher: Wiley
Correspondence: david.harvey@bioch.ox.ac.uk
Institutions: Department of Biochemistry, Oxford Glycobiology Institute, University of Oxford, Oxford OX1 3QU, UK
Methods: NMR, MALDI-TOF MS
- Article ID: 6301
Qin CJ, Ding MR, Tian GZ, Zou XP, Fu JJ, Hu J, Yin J "Chemical approaches towards installation of rare functional groups in bacterial surface glycans" -
Chinese Journal of Natural Medicines = Zhongguo Tianran Yaowu 20(6) (2022) 401-420
Bacterial surface glycans perform a diverse and important set of biological roles, and have been widely used in the treatment of bacterial infectious diseases. The majority of bacterial surface glycans are decorated with diverse rare functional groups, including amido, acetamidino, carboxamido and pyruvate groups. These functional groups are thought to be important constituents for the biological activities of glycans. Chemical synthesis of glycans bearing these functional groups or their variants is essential for the investigation of structure-activity relationships by a medicinal chemistry approach. To date, a broad choice of synthetic methods is available for targeting the different rare functional groups in bacterial surface glycans. This article reviews the structures of naturally occurring rare functional groups in bacterial surface glycans, and the chemical methods used for installation of these groups.
chemical synthesis, acetamidino group, amido group, bacterial surface glycan, carboxamido group, pyruvyl ketal
NCBI PubMed ID: 35750381Publication DOI: 10.1016/S1875-5364(22)60177-8Journal NLM ID: 101504416Publisher: Beijing: Science Press; Elsevier
Correspondence: J. Yin
Institutions: Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, China, Wuxi School of Medicine, Jiangnan University, Wuxi, China
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13. Compound ID: 2427
|
L-Orn-(5-6)-+ D-Asp-(1-4)-+
| |
b-D-GlcpNAc3NAcA-(1-4)-b-D-ManpNAc3NA-(1-3)-b-D-GlcpNAc-(1-3)-D-Fucp4N |
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Structure type: oligomer
Trivial name: repeating unit of the glycoconjugate polysaccharide part
Contained glycoepitopes: IEDB_135813,IEDB_137340,IEDB_141807,IEDB_151531
The structure is contained in the following publication(s):
- Article ID: 806
Hashimoto M, Asai Y, Jinno T, Adachi S, Kusumoto S "Structural elucidation of polysaccharide part of glycoconjugate from Treponema medium ATCC700293" -
European Journal of Biochemistry 270 (2003) 2671-2679
Glycoconjugates are distributed on the cell surfaces of some small-sized treponemes and have been reported to be completely different from lipopolysaccharides. We separated a glycoconjugate fraction from Treponema medium ATCC700293, a medium-sized oral spirochete, to assess its immunobiological activities and elucidate the chemical structure of its polysaccharide part using phenol/water extraction, hydrophobic chromatography, and gel .ltration. The glycoconjugate showed negligible or weak endotoxic and immunobiological properties. The chemical structure of the polysaccharide part was shown by two-dimensional NMR and MALDI-TOF-MS to be a tetrasaccharide backbone with two amino acids: -4)[Ac(1-3),Ac(1-2)]bDGlcpN3NA(1-4)[xLOrn(6-6)Ac(1-2)]bDManpN3NA(1-3)[Ac(1-2)]bDGlcpN(1-3)[xDAsp?(1-4)]aDFucp4N(1-.
structural, polysaccharide, aspartic acid, MALDI-TOF MS, ornithine, glycoconjugate, medium, Treponema
NCBI PubMed ID: 12787034Journal NLM ID: 0107600Publisher: Oxford, UK: Blackwell Science Ltd. on behalf of the Federation of European Biochemical Societies
Correspondence: tomo527@dent.asahi-u.ac.jp
Institutions: Department of Oral Microbiology, Asahi University School of Dentistry, Gifu, Japan, Graduate School of Science, Osaka University, Osaka, Japan
Methods: NMR-2D, ESI-MS, MALDI-TOF MS
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14. Compound ID: 2553
|
L-Ala2Fo-(1-3)-+
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-4)-b-D-GlcpNAc3NA6NH2-(1-4)-b-D-ManpNAc3NAcA6NH2-(1-4)-a-L-GulpNAc3NAcA-(1-3)-b-D-QuipNAc4NAc-(1- |
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Structure type: polymer chemical repeating unit
Trivial name: O-unit
Compound class: O-polysaccharide, O-antigen
The structure is contained in the following publication(s):
- Article ID: 872
Kocharova NA, Perepelov AV, Zatonsky GV, Shashkov AS, Knirel YA, Jansson P, Weintraub A "Structural studies of the O-specific polysaccharide of Vibrio cholerae O8 using solvolysis with triflic acid" -
Carbohydrate Research 330(1) (2001) 83-92
The O-specific polysaccharide (OPS) of Vibrio cholerae 08 was isolated by mild acid degradation of the lipopolysaccharide and studied by two-dimensional NMR spectroscopy, including NOESY and heteronuclear multiple-bond correlation (HMBC) experiments. The OPS was found to have a tetrasaccharide repeating unit with the following structure: →4)-β-D-Glcp NAc3NAcylAN-(1→4)-β-D-Manp NAc3NAcAN-(1→4)-α-L-Gulp NAc3NAcA-(1→3)-β-D-QuipNAc4NAc-(1→ where QuiNAc4NAc is 2,4-diacetamido-2,4,6-trideoxyglucose, GlcNAc3NAcylAN is 2-acetamido-3-(N-formyl-L-alanyl)amino-2,3-dideoxyglucuronamide, ManNAc3NAcAN is 2,3-diacetamido-2,3-dideoxymannuronamide, and GulNAc3NAcA is 2,3-diacetamido-2,3-dideoxyguluronic acid. The OPS was stable towards acid hydrolysis and solvolysis with anhydrous hydrogen fluoride, but could be cleaved selectively with trifluoromethanesulfonic (triflic) acid by the glycosidic linkages of β-QuiNAc4NAc and α-GulNAc3NAcA. The structures of the oligosaccharides obtained that were elucidated by electrospray ionization (ESI) MS and NMR spectroscopy, confirmed the OPS structure.
Lipopolysaccharide, O-specific polysaccharide, O-Specific polysaccharide structure, 2, 3-diamino-2, Vibrio cholerae, 3-dideoxyhexuronic acid, triflic acid solvolysis, solvolysis
NCBI PubMed ID: 11217966Publication DOI: 10.1016/S0008-6215(00)00271-8Journal NLM ID: 0043535Publisher: Elsevier
Correspondence: pererik.jansson@kfcmail.hs.sii.se
Institutions: Karolinska Institute, Clinical Research Center, Huddinge University Hospital, Huddinge, Sweden
Methods: NMR, HF solvolysis, ESI-MS, triflic acid solvolysis
- Article ID: 1452
Chatterjee SN, Chaudhuri K "Lipopolysaccharides of Vibrio cholerae. I. Physical and chemical characterization" -
Biochimica et Biophysica Acta 1639(2) (2003) 65-79
Vibrio cholerae is the causative organism of the disease cholera. The lipopolysaccharide (LPS) of V. cholerae plays an important role in eliciting the antibacterial immune response of the host and in classifying the vibrios into some 200 or more serogroups. This review presents an account of our up-to-date knowledge of the physical and chemical characteristics of the three constituents, lipid-A, core-polysaccharide (core-PS) and O-antigen polysaccharide (O-PS), of the LPS of V. cholerae of different serogroups including the disease-causing ones, O1 and O139. The structure and occurrence of the capsular polysaccharide (CPS) on V. cholerae O139 have been discussed as a relevant topic. Similarity and dissimilarity between the structures of LPS of different serogroups, and particularly between O22 and O139, have been analysed with a view to learning their role in the causation of the epidemic form of the disease by avoiding the host defence mechanism and in the evolution of the newer pathogenic strains in future. An idea of the emerging trends of research involving the use of immunogens prepared from synthetic oligosaccharides that mimic terminal epitopes of the O-PS of V. cholerae O1 in the development of a conjugate anti cholera vaccine is also discussed.
Lipopolysaccharide, structure, lipid A, capsular polysaccharide, serogroup, Vibrio cholerae
NCBI PubMed ID: 14559113Publication DOI: 10.1016/j.bbadis.2003.08.004Journal NLM ID: 0217513Publisher: Elsevier
Correspondence: sncac@sify.com (S.N. Chatterjee)
Institutions: Saha Institute of Nuclear Physics, 1/AF Bidhannagar, Sector-1, Calcutta-700 064, India, Biophysics Division, Indian Institute of Chemical Biology, Jadavpur, Calcutta-700 032, India
- Article ID: 1469
Knirel YA, Perepelov AV "Trifluoromethanesulfonic acid: a useful reagent for the solvolytic cleavage of glycosidic linkages in structural analysis of bacterial polysaccharides" -
Australian Journal of Chemistry 55(1-2) (2002) 69-72
The initial data on the application of triflic acid was summarized for the isolation of complex monosachharide derivatives and oligosachharides in the structural analysis of various bacterial polysachharides. Solvolysis proceeded selectively and without destruction of the sugars. Varying the temperature and duration of the reaction enabled preparation of oligosaccharides of different sizes.
polysaccharide, structural analysis, bacterial polysaccharides, solvolysis, trifluoromethanesulfonic acid, triflic acid
Publication DOI: 10.1071/CH01181Journal NLM ID: 0370614Correspondence: knirel@ioc.ac.ru
Institutions: N.D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia
Methods: 13C NMR, 1H NMR, GLC, MS, triflic acid solvolysis
- Article ID: 4329
Knirel YA "Structure of O-antigens" -
Book: Bacterial lipopolysaccharides: Structure, chemical synthesis, biogenesis and interaction with host cells (2011) Chapter 3, 41-115
The lipopolysaccharide (LPS) is the major constituent of the outer leaflet of the outer membrane of Gram-negative bacteria. Its lipid A moiety is embedded in the membrane and serves as an anchor for the rest of the LPS molecule. The outermost repetitive glycan region of the LPS is linked to the lipid A through a core oligosaccharide (OS), and is designated as the O-specific polysaccharide (O-polysaccharide, OPS) or O-antigen. The O-antigen is the most variable portion of the LPS and provides serological specificity, which is used for bacterial serotyping. The OPS also provides protection to the microorganisms from host defenses such as complement mediated killing and phagocytosis, and is involved in interactions of bacteria with plants and bacteriophages. Studies of the OPSs ranging from the elucidation of their chemical structures and conformations to their biological and physico-chemical properties help improving classification schemes of Gram-negative bacteria. Furthermore, these studies contributed to a better understanding of the mechanisms of pathogenesis of infectious diseases, as well as provided information to develop novel vaccines and diagnostic reagents.
Lipopolysaccharide, synthesis, lipopolysaccharides, structure, Bacterial, host, O-antigen, O antigen, cell, O antigens, O-antigens, chemical, interaction, cells, PDF, chemical synthesis, biogenesis
Publication DOI: 10.1007/978-3-7091-0733-1_3Publisher: Springer
Correspondence: knirel@ioc.ac.ru
Editors: Knirel YA, Valvano MA
Institutions: Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia
- Article ID: 5157
Goyette-Desjardins G, Vinogradov E, Okura M, Takamatsu D, Gottschalk M, Segura M "Streptococcus suis serotype 3 and serotype 18 capsular polysaccharides contain di-N-acetyl-bacillosamine" -
Carbohydrate Research 466 (2018) 18-29
Streptococcus suis serotype 3 is counted among the S. suis serotypes causing clinical disease in pigs. Yet, limited information is available on this serotype. Here we determined for the first time the chemical composition and structure of serotype 3 capsular polysaccharide (CPS), a major bacterial virulence factor and the antigen at the origin of S. suis classification into serotypes. Chemical and spectroscopic data gave the repeating unit sequence for serotype 3: [4)D-GlcA (β1-3)d-QuiNAc4NAc(β1-]n. To the best of our knowledge, this is the first report of di-N-acetyl-d-bacillosamine (QuiNAc4NAc) containing polysaccharides in Streptococci and the second time this rare diamino sugar has been observed in a Gram-positive bacterial species since its initial report. This led to the identification of homologues of UDP-QuiNAc4NAc synthesis genes in S. suis serotype 18. Thus, the repeating unit sequence for serotype 18 is: [3)d-GalNAc(α1-3)[d-Glc (β1-2)]d-GalA4OAc(β1-3)d-GalNAc(α1-3)d-QuiNAc4NAc(α1-]n. A correlation between S. suis serotypes 3 and 18 CPS sequences and genes of these serotypes' cps loci encoding putative glycosyltransferases and polymerase responsible for the biosynthesis of the repeating unit was tentatively established. Knowledge of CPS structure and composition will contribute to better dissect the role of this bacterial component in the pathogenesis of S. suis serotypes 3 and 18.
polysaccharide, capsular polysaccharide, polysaccharides, carbohydrate structure, Streptococcus suis, Di-N-Acetyl-bacillosamine, Serotype 18, Serotype 3
NCBI PubMed ID: 30014879Publication DOI: 10.1016/j.carres.2018.07.003Journal NLM ID: 0043535Publisher: Elsevier
Correspondence: mariela.segura@umontreal.ca
Institutions: Swine and Poultry Infectious Diseases Research Center, Faculty of Veterinary Medicine, University of Montreal, 3200 Sicotte St., St-Hyacinthe, Quebec, J2S 2M2, Canada, Canadian Glycomics Network (GlycoNet), University of Alberta, 11227 Saskatchewan Dr., Edmonton, Alberta, T6G 2G2, Canada, National Research Council, 100 Sussex Dr., Ottawa, Ontario, K1A 0R6, Canada, Division of Bacterial and Parasitic Disease, National Institute of Animal Health, National Agriculture and Food Research Organization, 3-1-5 Kannondai, Tsukuba, Ibaraki, 305-0856, Japan, The United Graduate School of Veterinary Sciences, Gifu University, 1-1 Yanagido, Gifu, Gifu, 501-1193, Japan
Methods: 13C NMR, 1H NMR, periodate oxidation, gel filtration, NMR-2D, GC-MS, de-O-acylation, sugar analysis, methanolysis, SEC-MALS, bioinformatic analysis
- Article ID: 6301
Qin CJ, Ding MR, Tian GZ, Zou XP, Fu JJ, Hu J, Yin J "Chemical approaches towards installation of rare functional groups in bacterial surface glycans" -
Chinese Journal of Natural Medicines = Zhongguo Tianran Yaowu 20(6) (2022) 401-420
Bacterial surface glycans perform a diverse and important set of biological roles, and have been widely used in the treatment of bacterial infectious diseases. The majority of bacterial surface glycans are decorated with diverse rare functional groups, including amido, acetamidino, carboxamido and pyruvate groups. These functional groups are thought to be important constituents for the biological activities of glycans. Chemical synthesis of glycans bearing these functional groups or their variants is essential for the investigation of structure-activity relationships by a medicinal chemistry approach. To date, a broad choice of synthetic methods is available for targeting the different rare functional groups in bacterial surface glycans. This article reviews the structures of naturally occurring rare functional groups in bacterial surface glycans, and the chemical methods used for installation of these groups.
chemical synthesis, acetamidino group, amido group, bacterial surface glycan, carboxamido group, pyruvyl ketal
NCBI PubMed ID: 35750381Publication DOI: 10.1016/S1875-5364(22)60177-8Journal NLM ID: 101504416Publisher: Beijing: Science Press; Elsevier
Correspondence: J. Yin
Institutions: Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, China, Wuxi School of Medicine, Jiangnan University, Wuxi, China
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15. Compound ID: 2554
|
L-Ala-(1-3)-+
|
-4)-b-D-GlcpNAc3NA6NH2-(1-4)-b-D-ManpNAc3NAcA6NH2-(1-4)-a-L-GulpNAc3NAcA-(1-3)-b-D-QuipNAc4NAc-(1- |
Show graphically |
Structure type: polymer chemical repeating unit
Compound class: O-polysaccharide
The structure is contained in the following publication(s):
- Article ID: 872
Kocharova NA, Perepelov AV, Zatonsky GV, Shashkov AS, Knirel YA, Jansson P, Weintraub A "Structural studies of the O-specific polysaccharide of Vibrio cholerae O8 using solvolysis with triflic acid" -
Carbohydrate Research 330(1) (2001) 83-92
The O-specific polysaccharide (OPS) of Vibrio cholerae 08 was isolated by mild acid degradation of the lipopolysaccharide and studied by two-dimensional NMR spectroscopy, including NOESY and heteronuclear multiple-bond correlation (HMBC) experiments. The OPS was found to have a tetrasaccharide repeating unit with the following structure: →4)-β-D-Glcp NAc3NAcylAN-(1→4)-β-D-Manp NAc3NAcAN-(1→4)-α-L-Gulp NAc3NAcA-(1→3)-β-D-QuipNAc4NAc-(1→ where QuiNAc4NAc is 2,4-diacetamido-2,4,6-trideoxyglucose, GlcNAc3NAcylAN is 2-acetamido-3-(N-formyl-L-alanyl)amino-2,3-dideoxyglucuronamide, ManNAc3NAcAN is 2,3-diacetamido-2,3-dideoxymannuronamide, and GulNAc3NAcA is 2,3-diacetamido-2,3-dideoxyguluronic acid. The OPS was stable towards acid hydrolysis and solvolysis with anhydrous hydrogen fluoride, but could be cleaved selectively with trifluoromethanesulfonic (triflic) acid by the glycosidic linkages of β-QuiNAc4NAc and α-GulNAc3NAcA. The structures of the oligosaccharides obtained that were elucidated by electrospray ionization (ESI) MS and NMR spectroscopy, confirmed the OPS structure.
Lipopolysaccharide, O-specific polysaccharide, O-Specific polysaccharide structure, 2, 3-diamino-2, Vibrio cholerae, 3-dideoxyhexuronic acid, triflic acid solvolysis, solvolysis
NCBI PubMed ID: 11217966Publication DOI: 10.1016/S0008-6215(00)00271-8Journal NLM ID: 0043535Publisher: Elsevier
Correspondence: pererik.jansson@kfcmail.hs.sii.se
Institutions: Karolinska Institute, Clinical Research Center, Huddinge University Hospital, Huddinge, Sweden
Methods: NMR, HF solvolysis, ESI-MS, triflic acid solvolysis
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