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1. Compound ID: 183
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R-3HOBut-(1-5)-a-Psep7Fo-(2-4)-b-D-Xylp-(1-3)-b-D-FucpNAc-(1-3)-L-Ser |
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Structure type: oligomer
Trivial name: pilin glycan
Contained glycoepitopes: IEDB_114701,IEDB_150900,IEDB_167188,IEDB_174332
The structure is contained in the following publication(s):
- Article ID: 38
Castric P, Cassels FJ, Carlson RW "Structural characterization of the Pseudomonas aeruginosa 1244 pilin glycan" -
Journal of Biological Chemistry 276(28) (2001) 26479-26485
An antigenic similarity between lipopolysaccharide (LPS) and glycosylated pilin of Pseudomonas aeruginosa 1244 was noted. We purified a glycan-containing molecule from proteolytically digested pili and showed it to be composed of three sugars and serine. This glycan competed with pure pili and LPS for reaction with an LPS-specific monoclonal antibody, which also inhibited twitching motility by P. aeruginosa bearing glycosylated pili. One-dimensional NMR analysis of the glycan indicated the sugars to be 5N β OHC(4)7NfmPse, Xyl, and FucNAc. The complete proton assignments of these sugars as well as the serine residue were determined by COSY and TOCSY. Electrospray ionization mass spectrometry (MS) determined the mass of this molecule to be 771.5. The ROESY NMR spectrum, tandem MS/MS analysis, and methylation analysis provided information on linkage and the sequence of oligosaccharide components. These data indicated that the molecule had the following structure: α-5N β OHC(4)7NFmPse-(2→4)β-Xyl-(1→3)-β-FucNAc-(1→3)-β-Ser
structural, characterization, Pseudomonas, Pseudomonas aeruginosa, glycan, pilin, pseudaminic acid
NCBI PubMed ID: 11342554Journal NLM ID: 2985121RPublisher: Baltimore, MD: American Society for Biochemistry and Molecular Biology
Correspondence: castric@duq.edu
Institutions: Department of Biological Sciences, Duquesne University, Pittsburgh, Pennsylvania 15282, USA
Methods: 1H NMR, GLC-MS, NMR-2D, sugar analysis, ESI-MS, Western blotting, amino acid analysis, MS/MS
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2. Compound ID: 185
Structure type: polymer chemical repeating unit
Compound class: O-polysaccharide, O-antigen, LPS
Contained glycoepitopes: IEDB_142488,IEDB_146664,IEDB_983931,SB_192
The structure is contained in the following publication(s):
- Article ID: 40
Dean CR, Franklund CV, Retief JD, Coyne MJ, Hatano K, Evans DJ, Pier GB, Goldberg JB "Characterization of the serogroup O11 O-antigen locus of Pseudomonas aeruginosa PA103" -
Journal of Bacteriology 181(14) (1999) 4275-4284
We previously cloned a genomic DNA fragment from the serogroup O11 Pseudomonas aeruginosa strain PA103 that contained all genes necessary for O-antigen synthesis and directed the expression of serogroup O11 antigen on recombinant Escherichia coli and Salmonella. To elucidate the pathway of serogroup O11 antigen synthesis, the nucleotide sequence of the biosynthetic genes was determined. Eleven open reading frames likely to be involved in serogroup O11 O-antigen biosynthesis were identified and are designated in order as wzzPaO111 (wzz from P. aeruginosa serogroup O11), wzxPaO11, wbjA, wzyPaO11, wbjB to wbjF, wbpLO11 and wbpMO11 (wbpL and wbpM from serogroup O11). Consistent with previous descriptions of O-antigen biosynthetic gene loci, the entire region with the exception of wbpMO11 has a markedly reduced G+C content relative to the chromosomal average. WzyPaO11 shows no significant similarity at the protein or DNA sequence level to any database sequence and is very hydrophobic, with 10 to 12 putative transmembrane domains, both typical characteristics of O-antigen polymerases. A nonpolar chromosomal insertion mutation in wzyPaO11 in P. aeruginosa PA103 confirmed the identity of this gene. There is striking similarity between WbjBCDE and Cap(5/8)EFGL, involved in type 5 and type 8 capsule biosynthesis in Staphylococcus aureus. There is nearly total identity between wbpMO11 and wbpMO5, previously shown by others to be present in all 20 P. aeruginosa serogroups. Using similarity searches, we have assigned functions to the proteins encoded by the PA103 O-antigen locus and present the potential steps in the pathway for the biosynthesis of P. aeruginosa serogroup O11 O antigen
biosynthesis, characterization, O-antigen, O antigen, Pseudomonas, Pseudomonas aeruginosa, locus, serogroup
NCBI PubMed ID: 10400585Journal NLM ID: 2985120RPublisher: American Society for Microbiology
Correspondence: jbg2b@virginia.edu
Institutions: Departments of Microbiology and Information Technology and Communications, University of Virginia Health Sciences Center, Charlottesville, Virginia 22908, USA, Channing Laboratory, Department of Medicine, Brigham and Women's Hospital and Harvard Medical School, Boston, Massachusetts 022153
- Article ID: 866
Kocharova NA, Knirel YA, Stanislavsky ES, Kholodkova EV, Lugowski C, Jachymek W, Romanowska E "Structural and serological studies of lipopolysaccharides of Citrobacter O35 and O38 antigenically related to Salmonella" -
FEMS Immunology and Medical Microbiology 13(1) (1996) 1-8
Structural analysis using 13C NMR spectroscopy and methylation showed that lipopolysaccharides (LPSs) of Citrobacter freundii O35 and Salmonella arizonae O59 have structurally identical O-specific polysaccharide chains, and those of C. freundii O38 and Salmonella kentucky differ only in the presence of O-acetyl groups in the former. Serological relationships between the structurally similar LPSs were demonstrated using inhibition of ELISA, rocket immunoelectrophoresis, double gel diffusion, and immunoblotting. The O-acetyl groups present in C. freundii O38 LPS are of little importance for its serological specificity. A cross-reaction was observed in immunoblotting between O-antisera to C. freundii O35 and S. arizonae O59 and a structurally related LPS of Pseudomonas aeruginosa O11a, 11b (Lanyi-Bergan classification).
Lipopolysaccharide, structure, Pseudomonas aeruginosa, O-specific polysaccharide, Salmonella, Citrobacter, immunospecificity, O-acetyl group
NCBI PubMed ID: 8821392Publication DOI: 10.1111/j.1574-695X.1996.tb00209.xJournal NLM ID: 9315554Publisher: Elsevier
Correspondence: knirel@ioc.ac.ru
Institutions: N.D.Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia
Methods: 13C NMR, methylation
- 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: 1808
Shashkov AS, Knirel YA, Kocharova NA, Dmitriev BA, Kochetkov NK "Carbon-13 NMR spectrum of Pseudomonas aeruginosa type 7a, b specific polysaccharide" -
Bioorganicheskaya Khimia = Bioorganic Chemistry [Russian] 6(9) (1980) 1332-1337
The 13C NR spectrum of the Pseudomonas aeruginosa O-serotype 7a, b specific polysaccharide was fully interpreted. The sequence of monosaccharide residues in the repeating oligosaccharide unit of the polysaccharide as well as the configuration of glycosidic linkages followed unequivocally from the comparative analysis of 13C NMR spectra of model fucosaminides, disaccharide fragment produced from the title polysaccharide by Smith's degradation and the polysaccharide. The spectral data confirmed the results of earlier structural investigation of the polysaccharide.
Journal NLM ID: 7804941WWW link: http://www.rjbc.ru/arc/6/9/1332-1337.pdfPublisher: Moskva: Nauka
Institutions: N.D. Zelinsky Institute of Organic Chemistry, Academy of Sciences of the USSR, Moscow, Russia
- 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: 2101
Lipkind GM, Shashkov AS, Knirel YA, Vinogradov EV, Kochetkov NK "A computer-assisted structural analysis of regular polysaccharides on the basis of 13C-NMR data" -
Carbohydrate Research 175(1) (1988) 59-75
A computerised approach to the structural analysis of unbranched regular polysaccharides is described, which is based on an evaluation of the 13C-n.m.r. spectra for all possible primary structures within the additive scheme starting from the chemical shifts of the 13C resonances of the constituent monosaccharides and the average values of the glycosylation effects. The analysis reveals a structure (or structures), the evaluated spectrum of which resembles most closely that observed. The approach has been verified by using a series of bacterial polysaccharides of known structure and, in combination with methylation analysis data, for the determination of the presently unknown structures of the O-specific polysaccharides from Salmonella arizonae O59 and O63, and Proteus hauseri O19.
NCBI PubMed ID: 3378242Journal NLM ID: 0043535Publisher: Elsevier
Institutions: N.D. Zelinsky Institute of Organic Chemistry, Academy of Sciences of the USSR, Moscow, Russia
Methods: 13C NMR
- Article ID: 2280
D'Ambra AJ, Gray GR "Analysis by the reductive-cleavage method of a polysaccharide containing 2-acetamido-2,6-dideoxy-D- and -L-galactopyranosyl residues" -
Carbohydrate Research 251 (1994) 127-144
Journal NLM ID: 0043535Publisher: Elsevier
- Article ID: 2361
Dmitriev BA, Knirel YA, Kocharova NA, Kochetkov NK, Stanislavsky ES, Mashilova GM "Somatic antigens of Pseudomonas aeruginosa. The structure of the polysaccharide chain of Ps. aeruginosa O-serogroup 7 (Lanyi) lipopolysaccharide" -
European Journal of Biochemistry 106 (1980) 643-651
A loosely bound lipopolysaccharide-protein complex was extracted from cells of Pseudomonas aeruginosa strain 170015 (O:7ab; Lanyi classification) by saline solution and purified from contaminant nucleic acid by Cetavlon treatment followed by precipitation in an ultracentrifuge. The saline-treated cells were re-extracted with hot aqueous phenol to give firmly bound lipopolysaccharide which was isolated from the phenol layer and purified by ultracentrifugaiton. The identity of both lipopolysaccharide preparations was proved by serological and chemical evidence. Mild acid degradation of the lipopolysaccharide resulted in the splitting off of a lipid component and led to polysaccharide which was purified by gel-filtration on a Sephadex G-50 column. The polysaccharide consisted of N-acetyl-D-fucosamine, N-acetyl-L-fucosamine and D-glucose in the ratio 1:1:1. On the basis of nuclear magnetic resonance spectra, results of methylation analysis and two sequential Smith degradations, the following structure can be assigned to the repeating unit of the polysaccharide: -3)LFucNAc(α 1-3)DFucNAc(β 1-2)DGlc(β 1-. The polysaccharide did not show serological activity whereas alkali-treated lipopolysaccharide readily sensitised sheep erythrocytes and inhibited the passive haemagglutination reaction with anti-(O:7a,b)serum. Evidence is presented that the oligosaccharide repeating units of the polysaccharide and alkali-treated lipopolysaccharide are indistinguishable. Ps. aeruginosa strain 170016 (O:7a,c) was shown to have the O-specific lipopolysaccharide identical with that from strain 170015. The presented data show that subfactors 7b and 7c in the Lanyi classification of Ps. aeruginosa O-antigens seem to relate to components of the bacterial surface other than lipopolysaccharides.
NCBI PubMed ID: 6772440Journal NLM ID: 0107600Publisher: Oxford, UK: Blackwell Science Ltd. on behalf of the Federation of European Biochemical Societies
Methods: 1H NMR
- Article ID: 2761
Knirel YA, Dmitriev BA, Kochetkov NK, Stanislavsky ES, Mashilova GM "O-Antigens of Pseudomonas aeruginosa: structures of the polysaccharide chains and immunochemical specificity" -
Molecular Genetics, Microbiology and Virology = Molekulyarnaya genetika, mikrobiologiya i virusologiya (1983) 13-18
Journal NLM ID: 8800525Publisher: New York, NY: Allerton Press
- Article ID: 3295
Yokota SI, Noguchi H "Epitopes for human monoclonal antibodies and serotyping antisera against the O-specific polysaccharide of Pseudomonas aeruginosa O11" -
Carbohydrate Research 261(1) (1994) 57-66
Epitopes for Pseudomonas aeruginosa O11-specific human monoclonal antibodies (mAbs) and O11 serotyping antisera have been characterized. These mAbs recognized the O-polysaccharide portion of the lipopolysaccharide. The structure of the O-polysaccharide of O11 has been reported to be comprised of trisaccharide repeating-units as follows: →3)-α-L-FucpNAc-(1→1)-β-D-FucpNAc-(1→ 2)-β-D-Glcp-(1→. (FucpNAc, 2-acetamido-2,6-dideoxygalactopyranoside.) Data from inhibition studies of binding in enzyme-linked immunosorbent assays and cell-agglutination assays, using monosaccharides and periodate-oxidized O-polysaccharide showed that the glucose residue, especially the C-3-C-6 segment and the β-anomeric configuration, in the polysaccharide is essential for the epitopes of all anti-O11 mAbs; however, the detailed epitope specificities were different from one another. Furthermore, epitopes for serotyping antisera of O11 seemed to be similar to those for the human mAbs.
Lipopolysaccharide, structure, human, polysaccharide, repeating unit, trisaccharide, Pseudomonas, Pseudomonas aeruginosa, Research, antibodies, antibody, epitope, monoclonal, monoclonal antibodies, monoclonal antibody, epitopes, MAb, O-polysaccharide, O polysaccharide, O-specific, O-specific polysaccharide, specificity, inhibition, glucose, serotyping, binding, monosaccharide, PDF, antiserum, assay, epitope specificity, configuration, enzyme-linked immunosorbent assay, immunosorbent, monosaccharides
NCBI PubMed ID: 7522126Publication DOI: 10.1016/0008-6215(94)80005-7Journal NLM ID: 0043535Publisher: Elsevier
Institutions: Sumitomo Pharmaceuticals Research Center, Osaka, Japan
Methods: periodate oxidation, gel filtration, ELISA, Western blotting, serological methods
- 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
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3. 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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4. Compound ID: 909
|
?%P-7)-D-gro-a-D-manHepp-(1-7)-L-gro-a-D-manHepp-(1-7)-+
|
b-D-Glcp-(1-3)-+ | b-D-Glcp-(1-4)-+ a-Kdop-(2-4)-+
| | | |
b-D-Glcp-(1-6)-a-D-GalpNAc-(1-6)-a-D-Galp-(1-4)-a-D-GalpNAc-(1-3)-b-D-FucpNAc-(1-3)-L-gro-a-D-manHepp-(1-3)-L-gro-a-D-manHepp-(1-5)-a-Kdop-(2--/lipid A/
|
b-D-Glcp-(1-2)-+ |
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Structure type: oligomer
Aglycon: lipid A
Compound class: core oligosaccharide
Contained glycoepitopes: IEDB_130648,IEDB_130650,IEDB_130659,IEDB_130670,IEDB_136906,IEDB_137472,IEDB_137473,IEDB_1391961,IEDB_140087,IEDB_140088,IEDB_140090,IEDB_141584,IEDB_141794,IEDB_142488,IEDB_144989,IEDB_146664,IEDB_151528,IEDB_190606,IEDB_2189046,IEDB_2189047,IEDB_226811,IEDB_885822,IEDB_983931,SB_192,SB_7
The structure is contained in the following publication(s):
- Article ID: 262
Holst O "On the occurrence of D-glycero-D-manno-heptose in lipopolysaccharides" -
Polish Journal of Chemistry 73 (1999) 1055-1067
Lipopolysaccharides (LPS) consist of three regions, i.e. the lipid A, the core region, and the O-specific polysaccharide. The core region and the lipid A represent a common structural unit occurring in all LPS. The structures of the core region of various bacteria have been investigated intensively for the past ten years, and several core regions containing D-glycero-D-manno-heptose which is the biosynthetic precursor of the common core constituent L-glycero-D-manno-heptose have been identified. In this review, these core structures are summarized and briefly discussed.
Lipopolysaccharide, lipopolysaccharides, core, D-glycero-D-manno-heptose, composition, occurrence
Journal NLM ID: 7901356WWW link: http://www.ichf.edu.pl/pjch/pj-1999/pj0799.htm#1055Publisher: Państwowe Wydawnictwo Naukowe
Institutions: Research Center Borstel, Center for Medicine and Biosciences, 23845 Borstel, Germany
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5. Compound ID: 962
|
-4)-b-D-ManpNAc3NAmA-(1-4)-b-D-ManpNAc3NAcA-(1-3)-b-D-FucpNAc4Ac-(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: 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
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-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
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7. Compound ID: 1191
Structure type: polymer chemical repeating unit
Compound class: CPS
Contained glycoepitopes: IEDB_240849
The structure is contained in the following publication(s):
- Article ID: 367
Sau S, Lee CY "Cloning of type 8 capsule genes and analysis of gene clusters for the production of different capsular polysaccharides in Staphylococcus aureus" -
Journal of Bacteriology 178(7) (1996) 2118-2126
Eleven serotypes of capsular polysaccharide from Staphylococcus aureus have been reported. We have previously cloned a cluster of type 1 capsule (cap1) genes responsible for type 1 capsular polysaccharide biosynthesis in S. aureus M. To clone the type 8 capsule (cap8) genes, a plasmid library of type 8 strain Becker was screened with a labelled DNA fragment containing the cap1 genes under low-stringency conditions. One recombinant plasmid containing a 14-kb insert was chosen for further study and found to complement 14 of the 18 type 8 capsule-negative (Cap8-) mutants used in the study. Additional library screening, subcloning, and complementation experiments showed that all of the 18 Cap8- mutants were complemented by DNA fragments derived from a 20.5-kb contiguous region of the Becker chromosome. The mutants were mapped into six complementation groups, indicating that the cap8 genes are clustered. By Southern hybridization analyses under high-stringency conditions, we found that DNA fragments containing the cap8 gene cluster show extensive homology with all 17 strains tested, including type 1 strains. By further Southern analyses and cloning of the cap8-related homolog from strain M, we show that strain M carries an additional capsule gene cluster different from the cap1 gene cluster. In addition, by using DNA fragments containing different regions of the cap8 gene cluster as probes to hybridize DNA from different strains, we found that the central region of the cap8 gene cluster hybridizes only to DNAs from certain strains tested whereas the flanking regions hybridize to DNAs of all strains tested. Thus, the cap8 gene clusters and its closely related homologs are likely to have organizations similar to those of the encapsulation genes of other bacterial systems.
capsular polysaccharides, analysis, cloning, type, gene cluster, capsule, Staphylococcus aureus
NCBI PubMed ID: 8606192Journal NLM ID: 2985120RPublisher: American Society for Microbiology
Correspondence: clee@kumc.edu
Institutions: Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, Kansas 66160
Methods: genetic methods
- Article ID: 584
Fattom AI, Horwith G, Fuller S, Propst M, Naso R "Development of StaphVAX, a polysaccharide conjugate vaccine against S. aureus infection: from the lab bench to phase III clinical trials" -
Vaccine 22(7) (2004) 880-887
Staphylococcus aureus is the most common nosocomial pathogen and is responsible for approximately one-third of hospital-acquired bacteremias. The emergence of strains with multidrug resistance, including resistance to vancomycin, the antibiotic of last resort, presents the medical community with a major public health problem. Alternative therapies, including immunotherapy, have been in development for several decades. The discovery of S. aureus capsular polysaccharides from clinical isolates, and their importance to pathogenicity via antiphagocytic activity, opened a new window of opportunity for development of vaccines and immunotherapy against this pathogen. A conjugate vaccine, StaphVAX that includes the two most prevalent capsular polysaccharides, types 5 and 8, coupled to a carrier protein efficient in promoting a Th2 response, was developed. In a recent phase III clinical study in hemodialysis patients, StaphVAX was shown to prevent S. aureus bacteremia for up to 10 months following a single immunization. The history, epidemiology, serology, and development of StaphVAX, including preclinical and clinical studies demonstrating efficacy are described in this review.
polysaccharide, Staphylococcus, conjugate vaccine
NCBI PubMed ID: 15040941Journal NLM ID: 8406899Publisher: Elsevier
Correspondence: afattom@nabi.com
Institutions: NABI Biopharmaceuticals, 12280 Wilkins Avenue, Rockville, MD 20852, USA
Methods: ELISA
- Article ID: 1374
Bhasin N, Albus A, Michon F, Livolsi PJ, Park JS, Lee JC "Identification of a gene essential for O-acetylation of the Staphylococcus aureus type 5 capsular polysaccharide" -
Molecular Microbiology 27(1) (1998) 9-21
gene, capsular, polysaccharide, capsular polysaccharide, type, O-acetylation, identification, Staphylococcus, Staphylococcus aureus
Journal NLM ID: 8712028Publisher: Blackwell Publishing
Correspondence: jean.lee@channing.harvard.edu
Institutions: Channing Laboratory, Department of Medicine, Brigham and Womens Hospital and Harvard Medical school, Boston, USA
- 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: 1520
Jones C "NMR assays for carbohydrate-based vaccines" -
Journal of Pharmaceutical and Biomedical Analysis 38(5) (2005) 840-850
Antibodies against the cell surface carbohydrates of many microbial pathogens protect against infection. This was initially exploited by the development of purified polysaccharide vaccines, but glycoconjugate vaccines, in which the cell surface carbohydrate of a microbial pathogen is covalently attached to an appropriate carrier protein, are proving the most effective means to generate this protective immunity. Carbohydrate-based vaccines against Haemophilus influenzae Type b, Neisseria meningitidis, Streptococcus pneumoniae and Salmonella enterica serotype Typhi (S. Typhi) are already licensed, and many similar products are in various stages of development. For many of these vaccines, biological assays are not available or are inappropriate and NMR spectroscopy is proving a valuable tool for the characterisation and quality control of existing and novel products. This review highlights some of the areas in which NMR spectroscopy is currently used, and where further developments may be expected.
capsular polysaccharide, O-acetylation, pneumonia, glycoconjugate, meningitis, carbohydrate-based vaccines, identity, typhoid
NCBI PubMed ID: 16087046Publication DOI: 10.1016/j.jpba.2005.01.044Journal NLM ID: 8309336Publisher: London: Elsevier
Institutions: Laboratory for Molecular Structure, National Institute for Biological Standards and Control, South Mimms, UK
Methods: NMR
- Article ID: 3296
Lee JC, Xu S, Albus A, Livolsi PJ "Genetic analysis of type 5 capsular polysaccharide expression by Staphylococcus aureus" -
Journal of Bacteriology 176(16) (1994) 4883-4889
Capsules are produced by over 90% of Staphylococcus aureus strains, and approximately 25% of clinical isolates express type 5 capsular polysaccharide (CP5). We mutagenized the type 5 strain Reynolds with Tn918 to target genes involved in CP5 expression. From a capsule-deficient mutant, we cloned into a cosmid vector an approximately 26-kb EcoRI fragment containing the transposon insertion. In the absence of tetracycline selection, Tn918 was spontaneously excised, thereby resulting in a plasmid containing 9.4 kb of S. aureus DNA flanking the Tn918 insertion site. The 9.4-kb DNA fragment was used to screen a cosmid library prepared from the wild-type strain. Positive colonies were identified by colony hybridization, and a restriction map of one clone (pJCL19 with an approximately 34-kb insert) carrying the putative capsule gene region was constructed. Fragments of pJCL19 were used to probe genomic DNA digests from S. aureus strains of different capsular serotypes. Fragments on the ends of the cloned DNA hybridized to fragments of similar sizes in most of the strains examined. Blots hybridized to two fragments flanking the central region of the cloned DNA showed restriction fragment length polymorphism. A centrally located DNA fragment hybridized only to DNA from capsular types 2, 4, and 5. DNA from pJCL19 was subcloned to a shuttle vector for complementation studies. A 6.2-kb EcoRI-ClaI fragment complemented CP5 expression in a capsule-negative mutant derived by mutagenesis with ethyl methanesulfonate. These experiments provide the necessary groundwork for identifying genes involved in CP5 expression by S. aureus.
genetic, clinical, expression, gene, genetics, DNA, strain, capsular, polysaccharide, serotype, analysis, capsular polysaccharide, type, mutant, region, insertion, plasmid, capsule, Staphylococcus, Staphylococcus aureus, fragment, Serotypes, site, clone, PDF, capsules, selection, polymorphism
NCBI PubMed ID: 805001Journal NLM ID: 2985120RPublisher: American Society for Microbiology
Institutions: Channing Laboratory, Department of Medicine, Brigham and Women's Hospital, Boston, Massachusetts.
Methods: serological methods, genetic methods
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8. Compound ID: 1192
Structure type: polymer chemical repeating unit
The structure is contained in the following publication(s):
- Article ID: 367
Sau S, Lee CY "Cloning of type 8 capsule genes and analysis of gene clusters for the production of different capsular polysaccharides in Staphylococcus aureus" -
Journal of Bacteriology 178(7) (1996) 2118-2126
Eleven serotypes of capsular polysaccharide from Staphylococcus aureus have been reported. We have previously cloned a cluster of type 1 capsule (cap1) genes responsible for type 1 capsular polysaccharide biosynthesis in S. aureus M. To clone the type 8 capsule (cap8) genes, a plasmid library of type 8 strain Becker was screened with a labelled DNA fragment containing the cap1 genes under low-stringency conditions. One recombinant plasmid containing a 14-kb insert was chosen for further study and found to complement 14 of the 18 type 8 capsule-negative (Cap8-) mutants used in the study. Additional library screening, subcloning, and complementation experiments showed that all of the 18 Cap8- mutants were complemented by DNA fragments derived from a 20.5-kb contiguous region of the Becker chromosome. The mutants were mapped into six complementation groups, indicating that the cap8 genes are clustered. By Southern hybridization analyses under high-stringency conditions, we found that DNA fragments containing the cap8 gene cluster show extensive homology with all 17 strains tested, including type 1 strains. By further Southern analyses and cloning of the cap8-related homolog from strain M, we show that strain M carries an additional capsule gene cluster different from the cap1 gene cluster. In addition, by using DNA fragments containing different regions of the cap8 gene cluster as probes to hybridize DNA from different strains, we found that the central region of the cap8 gene cluster hybridizes only to DNAs from certain strains tested whereas the flanking regions hybridize to DNAs of all strains tested. Thus, the cap8 gene clusters and its closely related homologs are likely to have organizations similar to those of the encapsulation genes of other bacterial systems.
capsular polysaccharides, analysis, cloning, type, gene cluster, capsule, Staphylococcus aureus
NCBI PubMed ID: 8606192Journal NLM ID: 2985120RPublisher: American Society for Microbiology
Correspondence: clee@kumc.edu
Institutions: Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, Kansas 66160
Methods: genetic methods
- Article ID: 584
Fattom AI, Horwith G, Fuller S, Propst M, Naso R "Development of StaphVAX, a polysaccharide conjugate vaccine against S. aureus infection: from the lab bench to phase III clinical trials" -
Vaccine 22(7) (2004) 880-887
Staphylococcus aureus is the most common nosocomial pathogen and is responsible for approximately one-third of hospital-acquired bacteremias. The emergence of strains with multidrug resistance, including resistance to vancomycin, the antibiotic of last resort, presents the medical community with a major public health problem. Alternative therapies, including immunotherapy, have been in development for several decades. The discovery of S. aureus capsular polysaccharides from clinical isolates, and their importance to pathogenicity via antiphagocytic activity, opened a new window of opportunity for development of vaccines and immunotherapy against this pathogen. A conjugate vaccine, StaphVAX that includes the two most prevalent capsular polysaccharides, types 5 and 8, coupled to a carrier protein efficient in promoting a Th2 response, was developed. In a recent phase III clinical study in hemodialysis patients, StaphVAX was shown to prevent S. aureus bacteremia for up to 10 months following a single immunization. The history, epidemiology, serology, and development of StaphVAX, including preclinical and clinical studies demonstrating efficacy are described in this review.
polysaccharide, Staphylococcus, conjugate vaccine
NCBI PubMed ID: 15040941Journal NLM ID: 8406899Publisher: Elsevier
Correspondence: afattom@nabi.com
Institutions: NABI Biopharmaceuticals, 12280 Wilkins Avenue, Rockville, MD 20852, USA
Methods: ELISA
- 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: 1520
Jones C "NMR assays for carbohydrate-based vaccines" -
Journal of Pharmaceutical and Biomedical Analysis 38(5) (2005) 840-850
Antibodies against the cell surface carbohydrates of many microbial pathogens protect against infection. This was initially exploited by the development of purified polysaccharide vaccines, but glycoconjugate vaccines, in which the cell surface carbohydrate of a microbial pathogen is covalently attached to an appropriate carrier protein, are proving the most effective means to generate this protective immunity. Carbohydrate-based vaccines against Haemophilus influenzae Type b, Neisseria meningitidis, Streptococcus pneumoniae and Salmonella enterica serotype Typhi (S. Typhi) are already licensed, and many similar products are in various stages of development. For many of these vaccines, biological assays are not available or are inappropriate and NMR spectroscopy is proving a valuable tool for the characterisation and quality control of existing and novel products. This review highlights some of the areas in which NMR spectroscopy is currently used, and where further developments may be expected.
capsular polysaccharide, O-acetylation, pneumonia, glycoconjugate, meningitis, carbohydrate-based vaccines, identity, typhoid
NCBI PubMed ID: 16087046Publication DOI: 10.1016/j.jpba.2005.01.044Journal NLM ID: 8309336Publisher: London: Elsevier
Institutions: Laboratory for Molecular Structure, National Institute for Biological Standards and Control, South Mimms, UK
Methods: NMR
- Article ID: 3296
Lee JC, Xu S, Albus A, Livolsi PJ "Genetic analysis of type 5 capsular polysaccharide expression by Staphylococcus aureus" -
Journal of Bacteriology 176(16) (1994) 4883-4889
Capsules are produced by over 90% of Staphylococcus aureus strains, and approximately 25% of clinical isolates express type 5 capsular polysaccharide (CP5). We mutagenized the type 5 strain Reynolds with Tn918 to target genes involved in CP5 expression. From a capsule-deficient mutant, we cloned into a cosmid vector an approximately 26-kb EcoRI fragment containing the transposon insertion. In the absence of tetracycline selection, Tn918 was spontaneously excised, thereby resulting in a plasmid containing 9.4 kb of S. aureus DNA flanking the Tn918 insertion site. The 9.4-kb DNA fragment was used to screen a cosmid library prepared from the wild-type strain. Positive colonies were identified by colony hybridization, and a restriction map of one clone (pJCL19 with an approximately 34-kb insert) carrying the putative capsule gene region was constructed. Fragments of pJCL19 were used to probe genomic DNA digests from S. aureus strains of different capsular serotypes. Fragments on the ends of the cloned DNA hybridized to fragments of similar sizes in most of the strains examined. Blots hybridized to two fragments flanking the central region of the cloned DNA showed restriction fragment length polymorphism. A centrally located DNA fragment hybridized only to DNA from capsular types 2, 4, and 5. DNA from pJCL19 was subcloned to a shuttle vector for complementation studies. A 6.2-kb EcoRI-ClaI fragment complemented CP5 expression in a capsule-negative mutant derived by mutagenesis with ethyl methanesulfonate. These experiments provide the necessary groundwork for identifying genes involved in CP5 expression by S. aureus.
genetic, clinical, expression, gene, genetics, DNA, strain, capsular, polysaccharide, serotype, analysis, capsular polysaccharide, type, mutant, region, insertion, plasmid, capsule, Staphylococcus, Staphylococcus aureus, fragment, Serotypes, site, clone, PDF, capsules, selection, polymorphism
NCBI PubMed ID: 805001Journal NLM ID: 2985120RPublisher: American Society for Microbiology
Institutions: Channing Laboratory, Department of Medicine, Brigham and Women's Hospital, Boston, Massachusetts.
Methods: serological methods, genetic methods
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9. Compound ID: 1205
|
D-gro-a-D-manHepp-(1-7)-L-gro-a-D-manHepp-(1-7)-+
|
b-D-Glcp-(1-3)-+ | b-D-Glcp-(1-4)-+
| | |
b-D-Glcp-(1-6)-a-D-GalpNAc-(1-6)-a-D-Galp-(1-4)-a-D-GalpNAc-(1-3)-b-D-FucpNAc-(1-3)-L-gro-a-D-manHepp-(1-3)-L-gro-a-D-manHepp-(1-5)-Kdop-(2--/lipid A/
|
b-D-Glcp-(1-2)-+ |
Show graphically |
Structure type: oligomer
Aglycon: lipid A
Compound class: core oligosaccharide
Contained glycoepitopes: IEDB_130648,IEDB_130650,IEDB_130670,IEDB_136906,IEDB_137472,IEDB_137473,IEDB_1391961,IEDB_140087,IEDB_140088,IEDB_140090,IEDB_141584,IEDB_141794,IEDB_142488,IEDB_144989,IEDB_146664,IEDB_151528,IEDB_190606,IEDB_2189046,IEDB_2189047,IEDB_226811,IEDB_885822,IEDB_983931,SB_192,SB_7
The structure is contained in the following publication(s):
- Article ID: 381
Skurnik M, Zhang L "Molecular genetics and biochemistry of Yersinia lipopolysaccharide" -
APMIS: Acta Pathologica, Microbiologica, et Immunologica Scandinavica 104(12) (1996) 849-872
Studies on the molecular genetics of bacterial LPS serve at least two main purposes: (i) to help develop an understanding of the biology, biochemistry and genetics of this bacterial surface macromolecule, and (ii) to provide a basis for both vaccine development and virulence experiments. Both of these goals have been the driving force in studies of Yersinia LPS carried out during the last decade. Here we will review the progress made in the molecular genetics and biochemistry of Yersinia LPS. A deep understanding has been achieved with respect to Y. enterocolitica serotype O:3, reaching as far as a detailed analysis of the gene clusters directing the biosynthesis of the outer core oligosaccharide and of the O-ag. The O-ag gene clusters of Y. enterocolitica serotype O:8 and Y. pseudotuberculosis serotypes O:2a and O:5a have also been cloned and partially characterized LPS biosynthesis of these Yersinia species includes examples of the two major variations recognized in the biosynthesis of this macromolecule: (i) homopolymeric or O-antigen polymerase-independent biosynthesis, and (ii) heteropolymeric or O-antigen polymerase-dependent biosynthesis.
Lipopolysaccharide, genetic, gene, genetics, O-antigen, biochemistry, Yersinia, molecular genetics
NCBI PubMed ID: 9048864Publication DOI: 10.1111/j.1699-0463.1996.tb04951.xJournal NLM ID: 8803400Publisher: Copenhagen: Munksgaard
Institutions: Turku Centre for Biotechnology, University of Turku, Finland, department of Medical Microbiology, University of Turku, Turku, Finland
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10. Compound ID: 1762
|
-3)-a-D-Quip4N-(1-4)-a-D-GlcpA-(1-6)-a-D-GlcpNAc-(1-4)-a-L-GulpNA-(1-3)-b-D-FucpNAc-(1- |
Show graphically |
Structure type: polymer chemical repeating unit
Compound class: O-polysaccharide
Contained glycoepitopes: IEDB_115136,IEDB_137340,IEDB_140630,IEDB_141807,IEDB_151531
The structure is contained in the following publication(s):
- Article ID: 547
Kilcoyne M, Perepelov AV, Tomshich SV, Komandrova NA, Shashkov AS, Romanenko LA, Knirel YA, Savage AV "Structure of the O-polysaccharide of Idiomarina zobellii KMM 231T containing two unusual amino sugars with the free amino group, 4-amino-4,6-dideoxy-D-glucose and 2-amino-2-deoxy-L-guluronic acid" -
Carbohydrate Research 339(3) (2004) 477-482
Mild acid degradation of the lipopolysaccharide of the bacterium Idiomarina zobellii, type strain KMM 231T, with aq 2% HOAc at 100 degrees C, yielded an oligosaccharide, which represents one repeating unit of the O-polysaccharide. A polysaccharide was obtained by mild base degradation of the lipopolysaccharide. The following structure of the O-polysaccharide was elucidated by 1H and 13C NMR spectroscopy of the oligosaccharide and base-degraded lipopolysaccharide, including COSY, TOCSY, ROESY, 1H, 13C HSQC, HSQC-TOCSY and HMBC experiments: [→3)-α-D-Quip4N-(1→4)-α-D-GlcpA-(1→6)-α-D-GlcpNAc-(1→4)-α-L-GulpNA-(1→3)-β-D-FucpNAc-(1→] The O-polysaccharide is distinguished by the presence of two unusual amino sugars, 4-amino-4,6-dideoxy-D-glucose (D-Qui4N) and 2-amino-2-deoxy-L-guluronic acid (L-GulNA), both having the free amino group. The unexpectedly high acid lability of the glycosidic linkage of 2-acetamido-2,6-dideoxy-D-galactose (D-FucNAc) could be associated with the presence of a free amino group adjacent to the site of attachment of FucNAc to Qui4N.
Lipopolysaccharide, 6-dideoxy-d-glucose, bacterial polysaccharide structure, 4-amino-4, Marine bacteria, Idiomarina zobellii, 2-Amino-2-deoxy-L-guluronic acid
NCBI PubMed ID: 15013384Journal NLM ID: 0043535Publisher: Elsevier
Correspondence: angela.savage@nuigalway.ie
Institutions: N.D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia, Department of Chemistry, National University of Ireland, Galway, Ireland, Pacific Institute of Bioorganic Chemistry, Far East Branch of the Russian Academy of Sciences, Vladivostok 690022, Russian Federation
Methods: methylation, NMR-2D, NMR, sugar analysis
- 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
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11. Compound ID: 1763
|
a-D-Quip4N-(1-4)-a-D-GlcpA-(1-6)-a-D-GlcpNAc-(1-4)-a-L-GulpNA-(1-3)-D-FucNAc |
Show graphically |
Structure type: oligomer
Compound class: O-polysaccharide
Contained glycoepitopes: IEDB_115136,IEDB_137340,IEDB_140630,IEDB_141807,IEDB_151531
The structure is contained in the following publication(s):
- Article ID: 547
Kilcoyne M, Perepelov AV, Tomshich SV, Komandrova NA, Shashkov AS, Romanenko LA, Knirel YA, Savage AV "Structure of the O-polysaccharide of Idiomarina zobellii KMM 231T containing two unusual amino sugars with the free amino group, 4-amino-4,6-dideoxy-D-glucose and 2-amino-2-deoxy-L-guluronic acid" -
Carbohydrate Research 339(3) (2004) 477-482
Mild acid degradation of the lipopolysaccharide of the bacterium Idiomarina zobellii, type strain KMM 231T, with aq 2% HOAc at 100 degrees C, yielded an oligosaccharide, which represents one repeating unit of the O-polysaccharide. A polysaccharide was obtained by mild base degradation of the lipopolysaccharide. The following structure of the O-polysaccharide was elucidated by 1H and 13C NMR spectroscopy of the oligosaccharide and base-degraded lipopolysaccharide, including COSY, TOCSY, ROESY, 1H, 13C HSQC, HSQC-TOCSY and HMBC experiments: [→3)-α-D-Quip4N-(1→4)-α-D-GlcpA-(1→6)-α-D-GlcpNAc-(1→4)-α-L-GulpNA-(1→3)-β-D-FucpNAc-(1→] The O-polysaccharide is distinguished by the presence of two unusual amino sugars, 4-amino-4,6-dideoxy-D-glucose (D-Qui4N) and 2-amino-2-deoxy-L-guluronic acid (L-GulNA), both having the free amino group. The unexpectedly high acid lability of the glycosidic linkage of 2-acetamido-2,6-dideoxy-D-galactose (D-FucNAc) could be associated with the presence of a free amino group adjacent to the site of attachment of FucNAc to Qui4N.
Lipopolysaccharide, 6-dideoxy-d-glucose, bacterial polysaccharide structure, 4-amino-4, Marine bacteria, Idiomarina zobellii, 2-Amino-2-deoxy-L-guluronic acid
NCBI PubMed ID: 15013384Journal NLM ID: 0043535Publisher: Elsevier
Correspondence: angela.savage@nuigalway.ie
Institutions: N.D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia, Department of Chemistry, National University of Ireland, Galway, Ireland, Pacific Institute of Bioorganic Chemistry, Far East Branch of the Russian Academy of Sciences, Vladivostok 690022, Russian Federation
Methods: methylation, NMR-2D, NMR, sugar analysis
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12. Compound ID: 1849
Structure type: polymer chemical repeating unit
Trivial name: CP5
Compound class: CPS, EPS, O-polysaccharide
Contained glycoepitopes: IEDB_240847,IEDB_240849
The structure is contained in the following publication(s):
- Article ID: 584
Fattom AI, Horwith G, Fuller S, Propst M, Naso R "Development of StaphVAX, a polysaccharide conjugate vaccine against S. aureus infection: from the lab bench to phase III clinical trials" -
Vaccine 22(7) (2004) 880-887
Staphylococcus aureus is the most common nosocomial pathogen and is responsible for approximately one-third of hospital-acquired bacteremias. The emergence of strains with multidrug resistance, including resistance to vancomycin, the antibiotic of last resort, presents the medical community with a major public health problem. Alternative therapies, including immunotherapy, have been in development for several decades. The discovery of S. aureus capsular polysaccharides from clinical isolates, and their importance to pathogenicity via antiphagocytic activity, opened a new window of opportunity for development of vaccines and immunotherapy against this pathogen. A conjugate vaccine, StaphVAX that includes the two most prevalent capsular polysaccharides, types 5 and 8, coupled to a carrier protein efficient in promoting a Th2 response, was developed. In a recent phase III clinical study in hemodialysis patients, StaphVAX was shown to prevent S. aureus bacteremia for up to 10 months following a single immunization. The history, epidemiology, serology, and development of StaphVAX, including preclinical and clinical studies demonstrating efficacy are described in this review.
polysaccharide, Staphylococcus, conjugate vaccine
NCBI PubMed ID: 15040941Journal NLM ID: 8406899Publisher: Elsevier
Correspondence: afattom@nabi.com
Institutions: NABI Biopharmaceuticals, 12280 Wilkins Avenue, Rockville, MD 20852, USA
Methods: ELISA
- Article ID: 1374
Bhasin N, Albus A, Michon F, Livolsi PJ, Park JS, Lee JC "Identification of a gene essential for O-acetylation of the Staphylococcus aureus type 5 capsular polysaccharide" -
Molecular Microbiology 27(1) (1998) 9-21
gene, capsular, polysaccharide, capsular polysaccharide, type, O-acetylation, identification, Staphylococcus, Staphylococcus aureus
Journal NLM ID: 8712028Publisher: Blackwell Publishing
Correspondence: jean.lee@channing.harvard.edu
Institutions: Channing Laboratory, Department of Medicine, Brigham and Womens Hospital and Harvard Medical school, Boston, USA
- Article ID: 1520
Jones C "NMR assays for carbohydrate-based vaccines" -
Journal of Pharmaceutical and Biomedical Analysis 38(5) (2005) 840-850
Antibodies against the cell surface carbohydrates of many microbial pathogens protect against infection. This was initially exploited by the development of purified polysaccharide vaccines, but glycoconjugate vaccines, in which the cell surface carbohydrate of a microbial pathogen is covalently attached to an appropriate carrier protein, are proving the most effective means to generate this protective immunity. Carbohydrate-based vaccines against Haemophilus influenzae Type b, Neisseria meningitidis, Streptococcus pneumoniae and Salmonella enterica serotype Typhi (S. Typhi) are already licensed, and many similar products are in various stages of development. For many of these vaccines, biological assays are not available or are inappropriate and NMR spectroscopy is proving a valuable tool for the characterisation and quality control of existing and novel products. This review highlights some of the areas in which NMR spectroscopy is currently used, and where further developments may be expected.
capsular polysaccharide, O-acetylation, pneumonia, glycoconjugate, meningitis, carbohydrate-based vaccines, identity, typhoid
NCBI PubMed ID: 16087046Publication DOI: 10.1016/j.jpba.2005.01.044Journal NLM ID: 8309336Publisher: London: Elsevier
Institutions: Laboratory for Molecular Structure, National Institute for Biological Standards and Control, South Mimms, UK
Methods: NMR
- Article ID: 1521
Jones C "Revised structures for the capsular polysaccharides from Staphylococcus aureus types 5 and 8, components of novel glycoconjugate vaccines" -
Carbohydrate Research 340(6) (2005) 1097-1106
Glycoconjugate vaccines based on the capsular polysaccharides (CPSs) from Staphylococcus aureus serotypes 5 and 8 conjugated to genetically detoxified recombinant exoprotein A (rEPA) from Pseudomonas aeruginosa have been shown, in Phase 3 clinical trials, to elicit a strong bactericidal immune response in end-stage renal disease patients. Such vaccines have the potential to reduce morbidity and mortality due to methicillin-resistant Staphylococcus aureus (MRSA), a major cause of hospital-acquired infection. The serotype 5 and 8 polysaccharides have been fully characterized by NMR spectroscopy and full structural analyses carried out. Published structures were found incorrect and the revised structures of the repeat units of the two polysaccharides are: [carbohydrate structure: see text]. Resonances indicative of the presence of peptidoglycan were observed in the spectra of both CPSs, consistent with reports that the CPS is covalently linked to peptidoglycan.
polysaccharide, NMR spectroscopy, capsule, Staphylococcus aureus, peptidoglycan, conjugate vaccine
NCBI PubMed ID: 15797125Journal NLM ID: 0043535Publisher: Elsevier
Institutions: Laboratory for Molecular Structure, National Institute for Biological Standards and Control, South Mimms, UK
Methods: NMR, sugar analysis, determination of absolute configuration
- Article ID: 2169
Moreau M, Richards JC, Fournier JM, Byrd RA, Karakawa WW, Vann WF "Structure of the type 5 capsular polysaccharide of Staphylococcus aureus" -
Carbohydrate Research 201(2) (1990) 285-297
The Staphylococcus aureus type 5 capsular polysaccharide is composed of 2-acetamido-2-deoxy-l-fucose (1 part), 2-acetamido-2-deoxy-d-fucose (1 part), and 2-acetamido-2-deoxy-d-mannuronic acid (1 part). On the basis of methylation analysis, optical rotation, high-field one- and two-dimensional 1H- and 13C-n.m.r. experiments, and selective cleavage with 70% aqueous hydrogen fluoride, the polysaccharide was found to be a partially O-acetylated (50%) polymer of the repeating trisaccharide unit, [→4)-3-O-Ac-β-d-ManpNAcA-(1→4)-α-l-FucpNAc-(1→3)-β-d-FucpNAc-(1→]n.
NCBI PubMed ID: 2224883Publication DOI: 10.1016/0008-6215(90)84244-OJournal NLM ID: 0043535Publisher: Elsevier
Institutions: Office of Biologics Research and Review, Food and Drug Administration, Bethesda, MD, USA, U.S.A., Division of Biological Sciences, National Research Council of Canada, Ottawa K1A OR6 Canada, Unité du Cholerá et des Vibrions, Institut Pasteur, F-75724 Paris France
Methods: 13C NMR, 1H NMR, methylation, GLC-MS, gel filtration, GLC, HF treatment
- Article ID: 3296
Lee JC, Xu S, Albus A, Livolsi PJ "Genetic analysis of type 5 capsular polysaccharide expression by Staphylococcus aureus" -
Journal of Bacteriology 176(16) (1994) 4883-4889
Capsules are produced by over 90% of Staphylococcus aureus strains, and approximately 25% of clinical isolates express type 5 capsular polysaccharide (CP5). We mutagenized the type 5 strain Reynolds with Tn918 to target genes involved in CP5 expression. From a capsule-deficient mutant, we cloned into a cosmid vector an approximately 26-kb EcoRI fragment containing the transposon insertion. In the absence of tetracycline selection, Tn918 was spontaneously excised, thereby resulting in a plasmid containing 9.4 kb of S. aureus DNA flanking the Tn918 insertion site. The 9.4-kb DNA fragment was used to screen a cosmid library prepared from the wild-type strain. Positive colonies were identified by colony hybridization, and a restriction map of one clone (pJCL19 with an approximately 34-kb insert) carrying the putative capsule gene region was constructed. Fragments of pJCL19 were used to probe genomic DNA digests from S. aureus strains of different capsular serotypes. Fragments on the ends of the cloned DNA hybridized to fragments of similar sizes in most of the strains examined. Blots hybridized to two fragments flanking the central region of the cloned DNA showed restriction fragment length polymorphism. A centrally located DNA fragment hybridized only to DNA from capsular types 2, 4, and 5. DNA from pJCL19 was subcloned to a shuttle vector for complementation studies. A 6.2-kb EcoRI-ClaI fragment complemented CP5 expression in a capsule-negative mutant derived by mutagenesis with ethyl methanesulfonate. These experiments provide the necessary groundwork for identifying genes involved in CP5 expression by S. aureus.
genetic, clinical, expression, gene, genetics, DNA, strain, capsular, polysaccharide, serotype, analysis, capsular polysaccharide, type, mutant, region, insertion, plasmid, capsule, Staphylococcus, Staphylococcus aureus, fragment, Serotypes, site, clone, PDF, capsules, selection, polymorphism
NCBI PubMed ID: 805001Journal NLM ID: 2985120RPublisher: American Society for Microbiology
Institutions: Channing Laboratory, Department of Medicine, Brigham and Women's Hospital, Boston, Massachusetts.
Methods: serological methods, genetic methods
- Article ID: 4431
Ovodov YS "Bacterial capsular antigens. Structural patterns of capsular antigens" -
Biochemistry (Moscow) 71(9) (2006) 937-954
Structural patterns of bacterial capsular antigens including capsular polysaccharides and exoglycans are given in this review. In addition, the immunological activity of capsular antigens and their role in type specificity of bacteria are discussed.
structure, capsular polysaccharides, bacterial capsular antigens, bacterial exoglycans, immunological activity, type specificity
NCBI PubMed ID: 17009947Publication DOI: 10.1134/S000629790609001XJournal NLM ID: 0376536Publisher: Nauka/Interperiodica
Correspondence: ovoys@physiol.komisc.ru
Institutions: Institute of Physiology, Komi Science Center, Urals Branch of the Russian Academy of Sciences, Syktyvkar 167982, Russia
- Article ID: 4884
Weidenmaier C, Lee JC "Structure and Function of Surface Polysaccharides of Staphylococcus aureus" -
Current Topics in Medicinal Chemistry (2016) 1-37
The major surface polysaccharides of Staphylococcus aureus include the capsular polysaccharide (CP), cell wall teichoic acid (WTA), and polysaccharide intercellular adhesin/poly-β(1-6)-N-acetylglucosamine (PIA/PNAG). These glycopolymers are important components of the staphylococcal cell envelope, but none of them is essential to S. aureus viability and growth in vitro. The overall biosynthetic pathways of CP, WTA, and PIA/PNAG have been elucidated, and the functions of most of the biosynthetic enzymes have been demonstrated. Because S. aureus CP and WTA (but not PIA/PNAG) utilize a common cell membrane lipid carrier (undecaprenyl-phosphate) that is shared by the peptidoglycan biosynthesis pathway, there is evidence that these processes are highly integrated and temporally regulated. Regulatory elements that control glycopolymer biosynthesis have been described, but the cross talk that orchestrates the biosynthetic pathways of these three polysaccharides remains largely elusive. CP, WTA, and PIA/PNAG each play distinct roles in S. aureus colonization and the pathogenesis of staphylococcal infection. However, they each promote bacterial evasion of the host immune defences, and WTA is being explored as a target for antimicrobial therapeutics. All the three glycopolymers are viable targets for immunotherapy, and each (conjugated to a carrier protein) is under evaluation for inclusion in a multivalent S. aureus vaccine. Future research findings that increase our understanding of these surface polysaccharides, how the bacterial cell regulates their expression, and their biological functions will likely reveal new approaches to controlling this important bacterial pathogen.
structure, Pathogenesis, capsular polysaccharide, polysaccharides, Staphylococcus aureus, Enzymes, teichoic acid, vaccine, surface polysaccharide, peptidoglycan biosynthesis
NCBI PubMed ID: 26728067Publication DOI: 10.1007/82_2015_5018Journal NLM ID: 101119673Publisher: Bentham Science Publishers
Correspondence: chrisweidenmaier@googlemail.com; JCLEE@BWH.HARVARD.EDU
Institutions: Interfaculty Institute for Microbiology and Infection Medicine Tubingen, University of Tubingen and German Center for Infection Research, Tubingen, Germany, Division of Infectious Diseases, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, USA
- Article ID: 4926
Sanapala SR, Kulkarni SS "Expedient Route To Access Rare Deoxy Amino L-Sugar Building Blocks for the Assembly of Bacterial Glycoconjugates" -
Journal of the American Chemical Society 138(14) (2016) 4938-4947
Bacterial glycoproteins and oligosaccharides contain several rare deoxy amino l-sugars which are virtually absent in the human cells. This structural difference between the bacterial and host cell surface glycans can be exploited for the development of carbohydrate based vaccines and target specific drugs. However, the unusual deoxy amino l-sugars present in the bacterial glycoconjugates are not available from natural sources. Thus, procurement of orthogonally protected rare l-sugar building blocks through efficient chemical synthesis is a crucial step toward the synthesis of structurally well-defined and homogeneous complex glycans. Herein, we report a general and expedient methodology to access a variety of unusual deoxy amino l-sugars starting from readily available l-rhamnose and l-fucose via highly regioselective, one-pot double serial and double parallel displacements of the corresponding 2,4-bistriflates using azide and nitrite anions as nucleophiles. Alternatively, regioselective monotriflation at O2, O3, and O4 of l-rhamnose/l-fucose allowed selective inversions at respective positions leading to diverse rare sugars. The orthogonally protected deoxy amino l-sugar building blocks could be stereoselectively assembled to obtain biologically relevant bacterial O-glycans, as exemplified by the first total synthesis of the amino linker-attached, conjugation-ready tetrasaccharide of O-PS of Yersinia enterocolitica O:50 strain 3229 and the trisaccharide of Pseudomonas chlororaphis subsp. aureofaciens strain M71.
trisaccharide, Pseudomonas, glycoconjugates, vaccines, L-rhamnose, L-fucose, Yersinia enterocolitica, chemical synthesis, O-glycans
NCBI PubMed ID: 27002789Publication DOI: 10.1021/jacs.6b01823Journal NLM ID: 7503056Publisher: American Chemical Society
Correspondence: suvarn@chem.iitb.ac.in
Institutions: Department of Chemistry, Indian Institute of Technology Bombay , Powai, Mumbai 400076, India
Methods: 13C NMR, 1H NMR, TLC, GLC, chemical synthesis, chemical methods, MS, glycosylation
- Article ID: 5079
Weidenmaier C, Lee JC "Structure and Function of Surface Polysaccharides of Staphylococcus aureus" -
Book: Staphylococcus aureus (series: Current Topics in Microbiology and Immunology) (2017) Vol. 409, 57-93
The major surface polysaccharides of Staphylococcus aureus include the capsular polysaccharide (CP), cell wall teichoic acid (WTA), and polysaccharide intercellular adhesin/poly-β(1-6)-N-acetylglucosamine (PIA/PNAG). These glycopolymers are important components of the staphylococcal cell envelope, but none of them is essential to S. aureus viability and growth in vitro. The overall biosynthetic pathways of CP, WTA, and PIA/PNAG have been elucidated, and the functions of most of the biosynthetic enzymes have been demonstrated. Because S. aureus CP and WTA (but not PIA/PNAG) utilize a common cell membrane lipid carrier (undecaprenyl-phosphate) that is shared by the peptidoglycan biosynthesis pathway, there is evidence that these processes are highly integrated and temporally regulated. Regulatory elements that control glycopolymer biosynthesis have been described, but the cross talk that orchestrates the biosynthetic pathways of these three polysaccharides remains largely elusive. CP, WTA, and PIA/PNAG each play distinct roles in S. aureus colonization and the pathogenesis of staphylococcal infection. However, they each promote bacterial evasion of the host immune defences, and WTA is being explored as a target for antimicrobial therapeutics. All the three glycopolymers are viable targets for immunotherapy, and each (conjugated to a carrier protein) is under evaluation for inclusion in a multivalent S. aureus vaccine. Future research findings that increase our understanding of these surface polysaccharides, how the bacterial cell regulates their expression, and their biological functions will likely reveal new approaches to controlling this important bacterial pathogen.
capsular polysaccharide, Staphylococcus aureus, teichoic acid, vaccine, cell wall teichoic acid
NCBI PubMed ID: 26728067Publication DOI: 10.1007/82_2015_5018Publisher: Cham: Springer.
Correspondence: JCLEE@BWH.HARVARD.EDU
Editors: Bagnoli F, Rappuoli R, Grandi G
Institutions: Interfaculty Institute for Microbiology and Infection Medicine Tubingen, University of Tubingen and German Center for Infection Research, Tubingen, Germany, Division of Infectious Diseases, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, USA
- Article ID: 5129
Berti F, De Ricco R, Rappuoli R "Role of O-Acetylation in the Immunogenicity of Bacterial Polysaccharide Vaccines" -
Molecules 23(6) (2018) 1340
The incidence of infectious diseases caused by several bacterial pathogens such as Haemophilus influenzae type b, Streptococcus pneumoniae, and Neisseria meningitidis, has been dramatically reduced over the last 25 years through the use of glycoconjugate vaccines. The structures of the bacterial capsular polysaccharide (CPS) antigens, extracted and purified from microbial cultures and obtained with very high purity, show that many of them are decorated by O-acetyl groups. While these groups are often considered important for the structural identity of the polysaccharides, they play a major role in the functional immune response to some vaccines such as meningococcal serogroup A and Salmonella typhi Vi, but do not seem to be important for many others, such as meningococcal serogroups C, W, Y, and type III Group B Streptococcus. This review discusses the O-acetylation status of CPSs and its role in the immunological responses of these antigens.
O-acetylation, Bacterial polysaccharide, conjugate vaccines, Bacterial Vaccines, carbohydrate antigens
NCBI PubMed ID: 29865239Publication DOI: 10.3390/molecules23061340Journal NLM ID: 100964009Publisher: Basel, Switzerland: MDPI
Correspondence: rino.x.rappuoli@gsk.com
Institutions: External R&D, GSK Vaccines, 53100 Siena, Italy, External R&D, GSK Vaccines, 53100 Siena, Ital
- Article ID: 5188
Micoli F, Costantino P, Adamo R "Potential targets for next generation anti-microbial glycoconjugate vaccines" -
FEMS Microbiology Reviews 42(3) (2018) 388-423
Cell surface carbohydrates have been proven optimal targets for vaccine development. Conjugation of polysaccharides to a carrier protein triggers a T-cell dependent immune response to the glycan moiety. Licensed glycoconjugate vaccines are produced by chemical conjugation of capsular polysaccharides to prevent meningitis caused by meningococcus, pneumococcus and Haemophilus influenzae type b. However, other classes of carbohydrates (O-antigens, exopolysaccharides, wall/teichoic acids) represent attractive targets for developing vaccines.Recent analysis from WHO/CHO underpins alarming concern towards antibiotic resistant bacteria, such as the so called ESKAPE pathogens (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa and Enterobacter spp.) and additional pathogens such as Clostridium difficile and Group A Streptococcus. Fungal infections are also becoming increasingly invasive for immunocompromised patients or hospitalized individuals. Other emergencies could derive from bacteria which spread during environmental calamities (Vibrio cholerae) or with potential as bioterrorism weapons (Burkholderia pseudomallei and mallei, Francisella tularensis). Vaccination could aid reducing the use of broad spectrum antibiotics and provide protection by herd immunity also to individuals who are not vaccinated.This review analyses structural and functional differences of the polysaccharides exposed on the surface of emerging pathogenic bacteria, combined with medical need and technological feasibility of corresponding glycoconjugate vaccines.
carbohydrates, glycoconjugates, vaccines, glycoengineering, antimicrobial resistance
NCBI PubMed ID: 29547971Publication DOI: 10.1093/femsre/fuy011Journal NLM ID: 8902526Publisher: Oxford University Press
Correspondence: Roberto Adamo
Institutions: GSK Vaccines Institute for Global Health (GVGH), Via Fiorentina 1, 53100 Siena
- Article ID: 5731
Behera A, Rai D, Kulkarni SS "Total Syntheses of Conjugation-Ready Trisaccharide Repeating Units of Pseudomonas aeruginosa O11 and Staphylococcus aureus Type 5 Capsular Polysaccharide for Vaccine Development" -
Journal of the American Chemical Society 142(1) (2020) 456-467
Pseudomonas aeruginosa belongs to the group of three 'critical priority' multi-drug-resistant pathogens listed by WHO and is responsible for severe and often deadly infections such as bloodstream infections and pneumonia. Staphylococcus aureus is also a 'high priority' pathogen which is a major cause of serious nosocomial infections such as bacteremia, sepsis, and endocarditis. Owing to their ability to adapt resistance to almost any antibiotics, vaccines against these pathogens are urgently required. These pathogens express structurally unique and densely functionalized glycans on their surfaces which are absent on the host cells. Such carbohydrate antigens are valuable targets for the development of glycoconjugate vaccines and diagnostics. Here, we report the first total synthesis of the conjugation-ready trisaccharide repeating unit of Pseudomonas aeruginosa O11 via a highly stereoselective and efficient assembly of a rare l-fucosamine- and d-fucosamine-containing 1,2-cis-linked disaccharide motif and its regioselective glycosylation at O3. A systematic study was conducted for the notoriously difficult glycosylation with the most unreactive axial 4-OH of the rare disaccharide, and the successful outcome was utilized to accomplish the total synthesis of an aminopropyl linker-attached trisaccharide repeating unit of Staphylococcus aureus capsular polysaccharide type 5, which is also a potential antigen for immunotherapy and vaccine development. The judicious selection of protecting groups and reaction conditions allowed the stereoselective assembly and selective functional group interconversions to access the structurally complex linker-attached trisaccharide repeating units, which are valuable tools for immunological evaluation and vaccine development. The strategy is useful for the synthesis of other structurally related complex glycans.
repeating unit, trisaccharide, Pseudomonas aeruginosa, capsular polysaccharide, Staphylococcus aureus, vaccine, chemical synthesis
NCBI PubMed ID: 31815459Publication DOI: 10.1021/jacs.9b11309Journal NLM ID: 7503056Publisher: American Chemical Society
Correspondence: Suvarn S. Kulkarni
Institutions: Department of Chemistry, Indian Institute of Technology Bombay, Powai, Mumbai 400076, India.
Methods: 13C NMR, 1H NMR, IR, TLC, chemical synthesis, chemical methods, glycosylation, HR-ESI-MS
- Article ID: 5790
Khanam A, Tiwari A, Mandal PK "Chiral auxiliaries: Usefullness in stereoselective glycosylation reactions and their synthetic applications" -
Carbohydrate Research 495 (2020) 108045
Oligosaccharides play a very important role in biological system and structure-activity relationships that is why it has a lot of application to medicinal chemistry and development of polysaccharide conjugate vaccines. The stereoselective introduction of a glycosidic linkage presents the principal challenge for biological importance oligosaccharide synthesis. The main aim of this review is to described the importance of chiral auxiliary and neibhouring group participation for the stereoselective 1,2-cis glycosidic bonds formation and their application in complex oligosaccharide synthesis.Numerous 1,2-cis-linked oligosaccharides and glyconjugates are naturally found in the compounds of blood group, human milk, antigens of bacterial lipopolysaccharide etc.that predominantly increased it's importance in this field.
Oligosaccharides, glycosylation, stereoselective, 1, 2-Cis glycosidic bonds, Chiral auxiliary
NCBI PubMed ID: 32679340Publication DOI: 10.1016/j.carres.2020.108045Journal NLM ID: 0043535Publisher: Elsevier
Correspondence: pk.mandal@cdri.res.in
Institutions: Academy of Scientific and Innovative Research, New Delhi, India, Medicinal and Process Chemistry Division, CSIR-Central Drug Research Institute, BS-10/1, Sector 10, Jankipuram Extension, Sitapur Road, P.O. Box 173, Lucknow, 226031, India.Medicinal and Process Chemistry Division, CSIR-Central Drug Research Institute, BS-10/1, Sector 10, Jankipuram Extension, Sitapur Road, P.O. Box 173, Lucknow, 226031, India
- Article ID: 5861
Visansirikul S, Kolodziej SA, Demchenko AV "Staphylococcus aureus capsular polysaccharides: a structural and synthetic perspective" -
Organic and Biomolecular Chemistry 18(5) (2020) 783-798
The S. aureus bacterium is surrounded by capsular polysaccharides. These capsular polysaccharides are important in the pathogenesis of staphylococcal infection. There are 11 serotypes of capsular polysaccharides that have been identified, and a majority of strains express capsular polysaccharides type 5 (CP5) or 8 (CP8). The main focus of this review is to describe recent advances in the area of the chemical synthesis of monosaccharide components of S. aureus CP, oligosaccharide assembly and functionalization. Chemical routes to obtain oligosaccharides derived from CP1, CP5 and CP8 represent a compendium of modern classics of the total synthesis of challenging glycan sequences.
synthesis, carbohydrates, Oligosaccharides, vaccines
NCBI PubMed ID: 31922180Publication DOI: 10.1039/c9ob02546dJournal NLM ID: 101154995Publisher: The Royal Society of Chemistry
Correspondence: demchenkoa@umsl.edu
Institutions: Department of Chemistry and Biochemistry, University of Missouri - St Louis, One University Boulevard, St Louis, MO 63121, USA, Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Mahidol University, 447 Sri-Ayuddhaya Road, Rajathevee, Bangkok, 10400, Thailand, Bioprocess R&D, Biotherapeutics Pharmaceutical Sciences, Pfizer, Inc., 875 Chesterfield Parkway W, Chesterfield, MO 63017, USA
- Article ID: 5868
Whitfield C, Wear SS, Sande C "Assembly of Bacterial Capsular Polysaccharides and Exopolysaccharides" -
Annual Review of Microbiology 74 (2020) 521-543
Polysaccharides are dominant features of most bacterial surfaces and are displayed in different formats. Many bacteria produce abundant long-chain capsular polysaccharides, which can maintain a strong association and form a capsule structure enveloping the cell and/or take the form of exopolysaccharides that are mostly secreted into the immediate environment. These polymers afford the producing bacteria protection from a wide range of physical, chemical, and biological stresses, support biofilms, and play critical roles in interactions between bacteria and their immediate environments. Their biological and physical properties also drive a variety of industrial and biomedical applications. Despite the immense variation in capsular polysaccharide and exopolysaccharide structures, patterns are evident in strategies used for their assembly and export. This review describes recent advances in understanding those strategies, based on a wealth of biochemical investigations of select prototypes, supported by complementary insight from expanding structural biology initiatives. This provides a framework to identify and distinguish new systems emanating from genomic studies
capsular polysaccharide, exopolysaccharide, extracellular polysaccharide, capsule, glycan biosynthesis, glycan export
NCBI PubMed ID: 32680453Publication DOI: 10.1146/annurev-micro-011420-075607Journal NLM ID: 0372370Correspondence: cwhitfie@uoguelph.ca
Institutions: Department of Molecular and Cellular Biology, University of Guelph, Guelph, Ontario N1G 2W1, Canada
- Article ID: 6211
Del Bino L, Osterlid KE, Wu DY, Nonne F, Romano MR, Codée J, Adamo R "Synthetic Glycans to Improve Current Glycoconjugate Vaccines and Fight Antimicrobial Resistance" -
Chemical Reviews 122(20) (2022) 15672-15716
Antimicrobial resistance (AMR) is emerging as the next potential pandemic. Different microorganisms, including the bacteria Acinetobacter baumannii, Clostridioides difficile, Escherichia coli, Enterococcus faecium, Klebsiella pneumoniae, Neisseria gonorrhoeae, Pseudomonas aeruginosa, non-typhoidal Salmonella, and Staphylococcus aureus, and the fungus Candida auris, have been identified by the WHO and CDC as urgent or serious AMR threats. Others, such as group A and B Streptococci, are classified as concerning threats. Glycoconjugate vaccines have been demonstrated to be an efficacious and cost-effective measure to combat infections against Haemophilus influenzae, Neisseria meningitis, Streptococcus pneumoniae, and, more recently, Salmonella typhi. Recent times have seen enormous progress in methodologies for the assembly of complex glycans and glycoconjugates, with developments in synthetic, chemoenzymatic, and glycoengineering methodologies. This review analyzes the advancement of glycoconjugate vaccines based on synthetic carbohydrates to improve existing vaccines and identify novel candidates to combat AMR. Through this literature survey we built an overview of structure-immunogenicity relationships from available data and identify gaps and areas for further research to better exploit the peculiar role of carbohydrates as vaccine targets and create the next generation of synthetic carbohydrate-based vaccines.
carbohydrates, glycan, glycoconjugate vaccine
NCBI PubMed ID: 35608633Publication DOI: 10.1021/acs.chemrev.2c00021Journal NLM ID: 2985134RPublisher: Chem Rev
Correspondence: J. Codée
; R. Adamo
Institutions: GSK, R&D, 53100 Siena, Italy, Leiden Institute of Chemistry, Leiden University, 2300 RA Leiden, The Netherlands
- Article ID: 6318
Srtefanetti G, Maclennan CA, Micoli F "Impact and Control of Sugar Size in Glycoconjugate Vaccines" -
Molecules 27(19) (2022) 6432
Glycoconjugate vaccines have contributed enormously to reducing and controlling encapsulated bacterial infections for over thirty years. Glycoconjugate vaccines are based on a carbohydrate antigen that is covalently linked to a carrier protein; this is necessary to cause T cell responses for optimal immunogenicity, and to protect young children. Many interdependent parameters affect the immunogenicity of glycoconjugate vaccines, including the size of the saccharide antigen. Here, we examine and discuss the impact of glycan chain length on the efficacy of glycoconjugate vaccines and report the methods employed to size polysaccharide antigens, while highlighting the underlying reaction mechanisms. A better understanding of the impact of key parameters on the immunogenicity of glycoconjugates is critical to developing a new generation of highly effective vaccines.
glycoconjugates, vaccine, immunogenicity, fragmentation of polysaccharides, sugar length
NCBI PubMed ID: 36234967Publication DOI: 10.3390/molecules27196432Journal NLM ID: 100964009Publisher: Basel, Switzerland: MDPI
Correspondence: G. Srtefanetti
Institutions: Department of Biomolecular Sciences, University of Urbino Carlo Bo, 61029 Urbino, Italy, Enteric and Diarrheal Diseases, Global Health, Bill & Melinda Gates Foundation, 500 5th Ave. N, Seattle, WA 98109, USA, The Jenner Institute, Nuffield Department of Medicine, University of Oxford, Oxford OX3 7DQ, UK, The Institute of Immunology and Immunotherapy, University of Birmingham, Birmingham B15 2TT, UK, GSK Vaccines Institute for Global Health, 53100 Siena, Italy
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13. Compound ID: 2454
Structure type: oligomer
Contained glycoepitopes: IEDB_2189047
The structure is contained in the following publication(s):
- Article ID: 838
Jachymek W, Niedziela T, Petersson C, Lugowski C, Czaja J, Kenne L "Structures of the O-specific polysaccharides from Yokenella regensburgei (Koserella trabulsii) strains PCM2476, 2477, 2478, and 2494: High-resolution magic-angle spinning NMR investigation of the O-specific polysaccharides in native lipopolysaccharides and directly on the surface of living bacteria" -
Biochemistry 38(36) (1999) 11788-11795
The structures of the carbohydrate O-specific side-chain moiety of the lipopolysaccharides (LPS) of Yokenella regensburgei, strains PCM2476, 2477, 2478, and 2494, have been investigated by 1H and 13C NMR, fast atom bombardment tandem mass spectrometry (FAB-MSMS), matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry, methylation analysis, partial acid hydrolysis, and immunological methods. It was concluded that the O-specific polysaccharides of strains 2476, 2477, 2478, and 2494 are composed of the same basic trisaccharide repeating unit having the structure →3)-a-D-FucpNAc-(1→2)-L-a-D-Hepp-(1→3)-6-deoxy-a-L-Talp-(1→, in which L-a-D-Hepp is L-glycero-a-D-manno-heptopyranose. The detailed analysis revealed, however, differences in O-acetylation patterns of the 6-deoxy-L-talose residue, with 2- and 4-O-acetyl disubstituted →3)-6-deoxy-R-L-Talp-(1→ in strain PCM2476 and a 2-O-acetylated residue in strains 2477, 2478, and 2494. These structures represent novel, trisaccharide repeating units of bacterial O-antigens that are characteristic and unique to the Y. regensburgei species. By use of the high-resolution magic-angle spinning (HR-MAS) technique, 1H NMR spectra of the O-polysaccharides directly in isolated LPS were obtained. This allowed for almost full assignment and structural determination of the polysaccharide. By this technique the O-polysaccharide components were also observed in their original form directly on the surface of living bacterial cells.
Lipopolysaccharide, NMR, lipopolysaccharides, structure, strain, polysaccharide, bacteria, O-specific, O-specific polysaccharide, polysaccharides, surface, O-specific polysaccharides, native, high-resolution, magic angle, high resolution
NCBI PubMed ID: 10512635Journal NLM ID: 0370623Publisher: American Chemical Society
Correspondence: Lennart.Kenne@kemi.slu.se
Institutions: Swedish University of Agricultural Sciences, Uppsala, Sweden, and L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland
Methods: methylation, partial acid hydrolysis, MALDI-TOF MS, FAB-MS/MS, HR-MAS NMR
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14. Compound ID: 3101
|
P-7)-D-gro-a-D-manHepp-(1-7)-L-gro-a-D-manHepp-(1-7)-+
|
b-D-Glcp-(1-3)-+ P-4)-+ | b-D-Glcp-(1-4)-+ a-Kdop-(2-4)-+
| | | | |
b-D-Glcp-(1-6)-a-D-GalpNAc-(1-6)-a-D-Galp-(1-4)-a-D-GalpNAc-(1-3)-b-D-FucpNAc-(1-3)-L-gro-a-D-manHepp-(1-3)-L-gro-a-D-manHepp-(1-5)-a-Kdop-(2--/lipid A/
|
b-D-Glcp-(1-2)-+ |
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Structure type: oligomer
Aglycon: lipid A
Compound class: core oligosaccharide
Contained glycoepitopes: IEDB_130648,IEDB_130650,IEDB_130659,IEDB_130670,IEDB_136906,IEDB_137472,IEDB_137473,IEDB_1391961,IEDB_140087,IEDB_140088,IEDB_140090,IEDB_141584,IEDB_141794,IEDB_142488,IEDB_144989,IEDB_146664,IEDB_151528,IEDB_190606,IEDB_2189046,IEDB_2189047,IEDB_226811,IEDB_885822,IEDB_983931,SB_192,SB_7
The structure is contained in the following publication(s):
- Article ID: 1124
Radziejewska-Lebrecht J, Skurnik M, Shashkov AS, Brade L, Rozalski A, Bartodziejska B, Mayer H "Immunochemical studies on R mutants of Yersinia enterocolitica O:3" -
Acta Biochimica Polonica 45(4) (1998) 1011-1019
Three mutants of Yersinia enterocolitica O:3, namely: YeO3-R1, YeO3-RfbR7 and YeO3-c-trs8-R were classified on the basis of sodium dodecyl sulphate/polyacrylamide gel electrophoresis (SDS/PAGE) profile of isolated lipopolysaccharides (LPS) as belonging to the Ra- (the first) and the Rc-type (the other two mutants). Methylation analysis, in addition to 13C and 1H NMR studies of purified core oligosaccharides revealed structures similar to those established previously for the full core of Y. enterocolitica O:3 in the case of the Ra mutant, and identical to that reported for the Rc mutant Ye75R, in the case of the two other mutants. The O-specific sugar, 6d-L-altrose, which forms a homopolymeric O-chain, was present in small amounts in all three LPS preparations, as well as in the core oligosaccha ride preparations along with the Ra and the Rc sugars, characteristic of the Y. enterocolitica O:3 core. This result is in line with genetic data, indicating that it is the inner core region which is the receptor for the O-specific chain in Y. enterocolitica O:3. This region seems likewise to be the anchoring region for the enterobacterial common antigen (ECA), as shown by SDS/PAGE/Western blot analysis with monoclonal antibodies against ECA. In addition, we also demonstrated that the Ye75R mutant Rc and its parental strain Ye75S, both were ECA-immunogenic strains. So far, ECA-immunogenic strains, i.e. those with LPS-linked ECA, were only identified in E. coli mutants of the R1, R4 and K-12 serotype.
lipopolysaccharide;enterobacterial common antigen;immunogenicity;R mutants;Yersinia enterocolitica
NCBI PubMed ID: 10397347Journal NLM ID: 14520300RPublisher: Panstwowe Wydawnictwo Naukowe
Correspondence: joaradle@us.edu.pl
Institutions: Department of Microbiology, University of Silesia, Katowice, Poland, Forschungszentrum Borstel, Borstel, Germany, Turku Centre for Biotechnology, University of Turku and Abo Akademi University, P.O. Box 123, FIN-20521 Turku, Finland, N.D. Zelinsky Institute of Organic Chemistry, Leninsky Prospect 47, Moscow, Russia, Institute of Microbiology and Immunology, University of Lodz, St. Banacha 12/16, 90-237 Losz, Poland, Max-Planck-Institut fur Immunobiologie, Stubeweg 51, D-79108 Freiburg, Germany
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15. Compound ID: 3194
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P-6)-+
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a-D-Glcp-(1-4)-+ |
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a-D-Glcp-(1-6)-+ | P-2)-+ |
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b-D-ManpNAc3NAmA-(1-4)-b-D-ManpNAc3NAcA-(1-3)-b-D-FucpNAc-(1-3)-a-L-Rhap-(1-3)-b-D-Glcp-(1-3)-a-D-GalpN-(1-3)-L-gro-a-D-manHepp7Cm-(1-3)-L-gro-a-D-manHepp-(1-5)-a-Kdop-(2--/lipid A/
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L-Ala-(1-2)-+ P-4)-+ |
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Structure type: oligomer
Aglycon: lipid A
Compound class: core oligosaccharide with O-unit
Contained glycoepitopes: IEDB_130650,IEDB_1330403,IEDB_136105,IEDB_137473,IEDB_140088,IEDB_142488,IEDB_144998,IEDB_146664,IEDB_189517,IEDB_2189047,IEDB_225177,IEDB_885823,IEDB_983931,SB_192
The structure is contained in the following publication(s):
- 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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