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1. Compound ID: 36
|
-2)-a-D-Rhap4NAc-(1-3)-a-L-Fucp-(1-4)-b-D-Glcp-(1-3)-a-D-GalpNAc-(1- |
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Structure type: suggested polymer biological repeating unit
Compound class: O-polysaccharide, O-antigen
Contained glycoepitopes: IEDB_130648,IEDB_136045,IEDB_137473,IEDB_1391961,IEDB_141584,IEDB_142488,IEDB_142489,IEDB_144562,IEDB_146664,IEDB_152214,IEDB_174333,IEDB_885822,IEDB_983931,SB_192,SB_86
The structure is contained in the following publication(s):
- Article ID: 15
Bilge SS, Vary JC, Dowell SF, Tarr PI "Role of the Escherichia coli O157:H7 O side chain in adherence and analysis of an rfb locus" -
Infection and Immunity 64 (1996) 4795-4801
Shiga-toxigenic Escherichia coli strains belonging to serotype O157 are important human pathogens, but the genetic basis of expression of the O157 antigen and the role played by the lipopolysaccharide O side chain in the adherence of this organism to epithelial cells are not understood. We performed TnphoA mutagenesis on E. coli O157:H7 strain 86-24 to identify a mutant (strain F12) deficient in O-antigen expression. Nucleotide sequence analysis demonstrated that the transposon inserted within an open reading frame with significant homology to rfbE of Vibrio cholerae O1 (U. H. Stroeher, L. E. Karageorgos, R. Morona, and P. A. Manning, Proc. Natl. Acad. Sci. USA 89:2566-2570, 1992), which is postulated to encode perosamine synthetase. This open reading frame was designated rfbE(EcO157:H7). The guanine-plus-cytosine fraction (0.35) suggests that rfbE(EcO157:H7) may have originated in a species other than E. coli. rfbE(EcO157:H7) is conserved in nontoxigenic E. coli O157 strains expressing a variety of other flagellar antigens but is not found in E. coli O55:H7 strains, which are more closely related to E. coli O157:H7. Strain F12 was significantly more adherent to HeLa cells in a quantitative adherence assay than was its E. coli O157:H7 parent, but they did not differ in other phenotypes. Restoration of the expression of the O side chain by complementation of the TnphoA mutation in strain F12 by a plasmid expressing intact rfbE(EcO157:H7) reduced the adherence of the hyperadherent strain F12. We conclude that rfbE(EcO157:H7) is necessary for the expression of the O157 antigen, that acquisition of E. coli rfb genes occurred independently in E. coli O157:H7 and unrelated O157 strains, and that the O side chain of E. coli O157:H7 lipopolysaccharide interferes with the adherence of E. coli O157:H7 to epithelial cells
Lipopolysaccharide, LPS, structure, expression, role, O-antigen, analysis, chain, side chain, adherence, Escherichia, Escherichia coli, Escherichia coli O157, Escherichia coli O157:H7, locus, O-side-chain, rfb, rfb locus
NCBI PubMed ID: 8890241Journal NLM ID: 0246127Publisher: American Society for Microbiology
Correspondence: tarr@u.washington.edu
Institutions: Department of Pediatrics, University of Washington School of Medicine, Seattle, USA
Methods: DNA sequencing, DNA cloning, SDS-PAGE
- Article ID: 295
Konadu EY, Parke JC, Tran HT, Bryla DA, Robbins JB, Szu SC "Investigational vaccine for Escherichia coli O157: Phase I study of O157 O-specific polysaccharide Pseudomonas aeruginosa recombinant exoprotein A conjugates in adults" -
Journal of Infectious Diseases 177(2) (1998) 383-387
Escherichia coli O157 causes severe enteritis and the extraintestinal complication hemolytic-uremic syndrome. Serum IgG against the surface polysaccharide antigen, the O-specific polysaccharide of lipopolysaccharide (LPS), may confer protective immunity by lysing the inocula. In a phase 1 clinical study, three investigational vaccines were studied in 87 healthy adults. The vaccines were prepared by covalently binding E. coli O157 O-specific polysaccharide with Pseudomonas aeruginosa recombinant exoprotein A. No significant reactions were reported. Most volunteers (81%) responded with a > 4-fold increase in IgG LPS antibodies 1 week after vaccination; all volunteers responded with a > 4-fold rise at 4 weeks and this level was sustained for 26 weeks after injection. All three vaccines elicited high titers of serum bactericidal activity that roughly correlated with the serum IgG and IgM LPS antibody levels. A phase 2 study in young children is planned.
phase, polysaccharide, Pseudomonas, Pseudomonas aeruginosa, Escherichia, Escherichia coli, Escherichia coli O157, O-specific, O-specific polysaccharide, O157, vaccine, Adult, conjugate, conjugates, recombinant
NCBI PubMed ID: 9466525Journal NLM ID: 0413675Publisher: Oxford: Oxford University Press
Correspondence: scszu@helix.nih.gov
Institutions: National Institutes of Health, Bethesda, Maryland, USA and Carolinas Medical Center, Charlotte, North Carolina, USA.
Methods: immunochemical methods
- Article ID: 407
Vinogradov E, Conlan JW, Perry MB "Serological cross-reaction between the lipopolysaccharide O-polysaccharide antigens of Escherichia coli O157:H7 and strains of Citrobacter freundii and Citrobacter sedlakii" -
FEMS Microbiology Letters 190(1) (2000) 157-161
A strain of Citrobacter sedlakii showing serological cross-reaction with Escherichia coli O157 antisera was demonstrated to produce a lipopolysaccharide O-antigen having an identical structure with that of the E. coli O157 O-antigen. A strain of Citrobacter freunndii showing similar cross-reaction with E. coli O157 specific monoclonal antibody was shown to produce a lipopolysaccharide O-antigen composed of a trisaccharide repeating unit having the structure [ 2)-α-D Rhap-(1-3)-β-D-Rhap-(1-4)-β-D-Glcp-(1-]. This O-antigen differs from that of the E. coli O157 O-antigen and also lacks a component 2-substituted 4-amino-4,6-dideoxy-α-D-mannopyranosyl residue implicated as the common epitope in the lipopolysaccharide O-antigens of previously investigated bacterial species showing serological cross-reactivity with E. coli O157 antisera. The C freundii O-antigen presents an interesting example of structural mimicry within a bacterial polysaccharide antigen.
Lipopolysaccharide, Escherichia coli O157:H7, antigens, O-polysaccharide, Citrobacter, Citrobacter freundii, cross-reaction
NCBI PubMed ID: 10981707Journal NLM ID: 7705721Publisher: Blackwell Publishing
Correspondence: evguenii.vinogradov@nrc.ca
Institutions: Institute for Biological Sciences, National Research Council, Ottawa, Ont. Canada, K1A 0R6
Methods: 13C NMR, 1H NMR, GLC-MS, NMR-2D, ELISA, GLC, immunodiffusion assays
- Article ID: 486
Samuel G, Hogbin JP, Wang L, Reeves PR "Relationships of the Escherichia coli O157, O111, and O55 O-antigen gene clusters with those of Salmonella enterica and Citrobacter freundii, which express identical O antigens" -
Journal of Bacteriology 186(19) (2004) 6536-6543
Escherichia coli O157, Salmonella enterica O30, and Citrobacter freundii F90 have identical O-antigen structures, as do E. coli O55 and S. enterica O50. The O-antigen gene cluster sequences for E. coli O157 and E. coli O55 have been published, and the genes necessary for O-antigen biosynthesis have been identified, although transferase genes for glycosidic linkages are only generic and have not been allocated to specific linkages. We determined sequences for S. enterica O30 and C. freundii F90 O-antigen gene clusters and compared them to the sequence of the previously described E. coli O157 cluster. We also determined the sequence of the S. enterica O50 O-antigen gene cluster and compared it to the sequence of the previously described E. coli O55 cluster. For both the S. enterica O30-C. freundii F90-E. coli O157 group and the S. enterica O50-E. coli O55 group of O antigens, the gene clusters have identical or nearly identical organizations. The two sets of gene clusters had comparable overall levels of similarity in their genes, which were lower than the levels determined for housekeeping genes for these species, which were 55 to 65% for the genes encoding glycosyltransferases and O-antigen processing proteins and 75 to 93% for the nucleotide-sugar pathway genes. Nonetheless, the similarity of the levels of divergence in the five gene clusters required us to consider the possibility that the parent gene cluster for each structure was in the common ancestor of the species and that divergence is faster than expected for the common ancestor hypothesis. We propose that the identical O-antigen gene clusters originated from a common ancestor, and we discuss some possible explanations for the increased rate of divergence that is seen in these genes.
structure, genetics, Escherichia coli, Escherichia coli O157, Molecular Sequence Data, gene cluster, O-antigens, glycosyltransferases, Salmonella enterica, Citrobacter freundii
NCBI PubMed ID: 15375135Journal NLM ID: 2985120RPublisher: American Society for Microbiology
Correspondence: p.reeves@angis.usyd.edu.au
Institutions: School of Molecular and Microbial Biosciences, University of Sydney, NSW 2006, Australia
- Article ID: 1707
Perry MB, MacLean L, Griffith DW "Structure of the O-chain polysaccharide of the phenol-phase soluble lipopolysaccharide of Escherichia coli O:157:H7" -
Biochemistry and Cell Biology 64 (1986) 21-28
The phenol-phase soluble lipopolysaccharide isolated from Escherichia coli 0:157 by the hot phenol-water extraction procedure was shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, periodate oxidation, methylation, and 13C and 1H nuclear magnetic resonance studies to be an unbranched linear polysaccharide with a tetrasaccharide repeating unit having the structure: (formula; see text) The serological cross-reactivity of E. coli 0:157 with Brucella abortus, Yersinia enterocolitica (serotype 0:9), group N Salmonella, and some other E. coli species can be related immunochemically to the presence of 1,2-glycosylated N-acylated 4-amino-4, 6-dideoxy-α-D-mannopyranosyl residues in the O-chains of their respective lipopolysaccharides.
NCBI PubMed ID: 3008786Journal NLM ID: 8606068Publisher: Ottawa: National Research Council of Canada
Institutions: Division of Biological Sciences, National Research Council of Canada, Ottawa, Ont., Canada KIA OR6
Methods: 13C NMR, 1H NMR
- 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: 3197
Stenutz R, Weintraub A, Widmalm G "The structures of Escherichia coli O-polysaccharide antigens" -
FEMS Microbiology Reviews 30(3) (2006) 382-403
Escherichia coli is usually a non-pathogenic member of the human colonic flora. However, certain strains have acquired virulence factors and may cause a variety of infections in humans and in animals. There are three clinical syndromes caused by E. coli: (i) sepsis/meningitis; (ii) urinary tract infection and (iii) diarrhoea. Furthermore the E. coli causing diarrhoea is divided into different 'pathotypes' depending on the type of disease, i.e. (i) enterotoxigenic; (ii) enteropathogenic; (iii) enteroinvasive; (iv) enterohaemorrhagic; (v) enteroaggregative and (vi) diffusely adherent. The serotyping of E. coli based on the somatic (O), flagellar (H) and capsular polysaccharide antigens (K) is used in epidemiology. The different antigens may be unique for a particular serogroup or antigenic determinants may be shared, resulting in cross-reactions with other serogroups of E. coli or even with other members of the family Enterobacteriacea. To establish the uniqueness of a particular serogroup or to identify the presence of common epitopes, a database of the structures of O-antigenic polysaccharides has been created. The E. coli database (ECODAB) contains structures, nuclear magnetic resonance chemical shifts and to some extent cross-reactivity relationships. All fields are searchable. A ranking is produced based on similarity, which facilitates rapid identification of strains that are difficult to serotype (if known) based on classical agglutinating methods. In addition, results pertinent to the biosynthesis of the repeating units of O-antigens are discussed. The ECODAB is accessible to the scientific community at http://www.casper.organ.su.se/ECODAB/
NMR, structure, serotype, O-antigen, Enterobacteriacea, database
NCBI PubMed ID: 16594963Publication DOI: 10.1111/j.1574-6976.2006.00016.xJournal NLM ID: 8902526Publisher: Oxford University Press
Correspondence: andrej.weintraub@ki.se
Institutions: Department of Organic Chemistry, Arrhenius Laboratory, Stockholm University, Stockholm, Sweden
- Article ID: 4411
Vinnikova AN, Druzhinina TN, Danilov LL, Utkina NS, Torgov VI, Veselovsky VV, Wang S, Liu B, Wang L, Brockhausen I "Synthesis of a fluorescent acceptor substrate for glycosyltransferases involved in the assembly of O-antigens of enterohemorrhagic Escherichia coli O157 and O5" -
Carbohydrate Research 366 (2013) 17-24
The assembly of the repeating units of O-antigens in Gram negative bacteria is catalyzed by specific glycosyltransferases. Previously we used GlcNAc/GalNAc?-diphosphate-phenoxyundecyl as natural acceptor substrate analogs in assays of the transfer of radioactive sugars by bacterial glycosyltransferases. In order to develop new, fluorescence based assays we have synthesized a fluorescent acceptor P(1)-[11-(anthracen-9-ylmethoxy)undecyl]-P(2)-(2-acetamido-2-deoxy-α-d-galactopyranosyl) diphosphate and have shown that the compound was an excellent acceptor for glucosyltransferase WbdN from Escherichia coli (E. coli) O157 and for galactosyltransferase WbwC from E. coli O5. This is the first report of the Gal-transferase activity of the wbwC gene product of E.coli O5. The presence of the fluorescent label in the acceptor molecule allows the detection of glycosyltransferase reaction products with high sensitivity, eliminating the need for radioactive nucleotide sugars.
Escherichia coli O157, glycosyltransferases, chemical synthesis, WbdN, fluorescent assay, E. coli O157, E. coli O5
NCBI PubMed ID: 23261778Publication DOI: 10.1016/j.carres.2012.11.009Journal NLM ID: 0043535Publisher: Elsevier
Correspondence: I. Brockhausen
Institutions: N.D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia, Department of Medicine and Department of Biomedical and Molecular Sciences, Queen's University, Kingston, Canada
Methods: 13C NMR, 1H NMR, glycosyltransferase assays, 31P NMR, MALDI-MS, chemical methods, biochemical methods, HPLC, TCL
- Article ID: 5423
Dobrochaeva KL, Khasbiulina NR, Shilova NV, Obukhova PS, Knirel YA, Nokel AY, Bovin NV "Human antibodies eluted from ligand-free Sepharose capable of binding bacterial polysaccharides and sulfated glycans" -
Molecular Immunology 106 (2019) 63-68
Sepharose matrix without immobilized ligands binds antibodies from human blood serum or immunoglobulin preparations. The eluted antibodies bind bacterial polysaccharides having no structural similarity to agarose (Sepharose is a cross-linked polysaccharide agarose) with a high affinity. It is concluded that the identified antibodies are capable of recognizing spatial rather than linear epitopes of bacterial polysaccharides. This side activity of Sepharose matrix should be taken into account in isolating target antibodies and other proteins from human blood.
antibodies, bacteria, polysaccharides, printed glycan array, Agarose, Sepharose
NCBI PubMed ID: 30583222Publication DOI: 10.1016/j.molimm.2018.12.011Journal NLM ID: 7905289Publisher: Elsevier
Correspondence: N.V. Bovin
Institutions: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 16/10 ul. Miklukho-Maklaya, Moscow, Russian Federation, Semiotik LLC, Moscow, Russian Federation, N. D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, 47 Leninsky pr., Moscow, Russian Federation, School of Engineering, Computer & Mathematical Sciences, Auckland University of Technology, Auckland 1010, New Zealand
Methods: serological methods, UV, affinity chromatography, antibody binding, glycan array analysis, microarray binding assays, serum analysis, isolation of antibodies
- Article ID: 6430
Melamed J, Brockhausen I "Biosynthesis of the O antigen of pathogenic Escherichia coli O157:H7. Characterization of α1,4-Fuc-transferase WbdO" -
Glycobiology 33(2) (2023) 165-175
The O157:H7 strain of Escherichia coli is responsible for frequent outbreaks of hemorrhagic colitis worldwide. Its lipopolysaccharide is a virulence factor and contains an O antigen having repeating units with the tetrasaccharide structure [2-D-PerNAcα1-3-L-Fucα1-4-D-Glcβ1-3-D-GalNAcα1-]n. Genes encoding glycosyltransferases WbdN, WbdO, and WbdP are responsible for the biosynthesis of this repeating unit. We have previously characterized the second enzyme in the pathway, WbdN, which transfers Glc in β1-3 linkage to GalNAcα-O-PO3-PO3-(CH2)11-O-Ph (GalNAc-PP-PhU). In this work, Fuc-transferase WbdO from E. coli O157:H7 expressed in BL21 bacteria was characterized using the product of WbdN as the acceptor substrate. We showed that WbdO is specific for GDP-β-L-Fuc as the donor substrate. Compounds that contained terminal Glc or Glcβ1-3GalNAc structures but lacked the diphosphate group did not serve as acceptor substrates. The structure of the WbdO product was identified by mass spectrometry and Nuclear magnetic resonance (NMR) as L-Fucα1-4-D-Glcβ1-3-D-GalNAc PP-PhU. WbdO is an unusual bivalent metal ion-dependent Fuc-transferase classified as an inverting GT2 family enzyme that has 2 conserved sequences near the N-terminus. The Asp37 residue within the 36VDGGSTD42 sequence was found to be essential for catalysis. Mutation of Asp68 to Ala within the conserved 67YDAMNK72 sequence resulted in a 3-fold increase in activity. These studies show that WbdOO157 is a highly specific Fuc-transferase with little homology to other characterized Fuc-transferases.
O antigen, Escherichia coli O157:H7, specificity, fucosyltransferase, WbdO
NCBI PubMed ID: 36715215Publication DOI: 10.1093/glycob/cwac079Journal NLM ID: 9104124Publisher: IRL Press at Oxford University Press
Correspondence: I. Brockhausen
Institutions: Department of Biomedical and Molecular Sciences, Queen's University, 18 Stuart Street, Kingston, ON K7L3N6, Canada
Methods: 13C NMR, 1H NMR, gel filtration, NMR-2D, ESI-MS, Western blotting, radiolabeling, genetic methods, biochemical methods, HPLC, bioinformatic analysis, crystallization, enzymatic assays, chemoenzymatic synthesis, protein expression
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2. Compound ID: 178
|
-2)-a-D-Rhap4NAc-(1-3)-a-L-Fucp-(1-4)-b-D-Glcp-(1-3)-a-D-GalpNAc-(1- |
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Structure type: polymer chemical repeating unit
Compound class: O-polysaccharide, O-antigen
Contained glycoepitopes: IEDB_130648,IEDB_136045,IEDB_137473,IEDB_1391961,IEDB_141584,IEDB_142488,IEDB_142489,IEDB_144562,IEDB_146664,IEDB_152214,IEDB_174333,IEDB_885822,IEDB_983931,SB_192,SB_86
The structure is contained in the following publication(s):
- Article ID: 37
Caroff M, Karibian D "Structure of bacterial lipopolysaccharides" -
Carbohydrate Research 338(23) (2003) 2431-2447
Bacterial lipopolysaccharides are the major components of the outer surface of Gram-negative bacteria They are often of interest in medicine for their immunomodulatory properties. In small amounts they can be beneficial, but in larger amounts they may cause endotoxic shock. Although they share a common architecture, their structural details exert a strong influence on their activity. These molecules comprise: a lipid moiety, called lipid A, which is considered to be the endotoxic component, a glycosidic part consisting of a core of approximately 10 monosaccharides and, in 'smooth-type' lipopolysaccharides, a third region, named O-chain, consisting of repetitive subunits of one to eight monosaccharides responsible for much of the immunospecificity of the bacterial cell.
Lipopolysaccharide, structure, core, lipid A, endotoxin, O-chains
NCBI PubMed ID: 14670707Publication DOI: 10.1016/j.carres.2003.07.010Journal NLM ID: 0043535Publisher: Elsevier
Correspondence: martine.carloff@bbmpc.u-psud.fr
Institutions: Equipe Endotoxines, UMR 8619 du Centre National de la Recherche Scientifique, IBBMC, Université de Paris-Sud, F-Orsay, France
- Article ID: 1059
Nishiuchi Y, Doe M, Hotta H, Kobayashi K "Addendum to: 'Structure and serological properties of O-specific polysaccharide from Citrobacter freundii possessing cross-reactivity with Escherichia coli O157:H7'. [FEMS Immunol. Med. Microbiol. 28 (2000) 163-171]" -
FEMS Immunology and Medical Microbiology 32(3) (2002) 255
structure, polysaccharide, property, Escherichia, Escherichia coli, O-specific, O-specific polysaccharide, serological, cross-reactivity, crossreactivity, Citrobacter, Citrobacter freundii
NCBI PubMed ID: 11934572Journal NLM ID: 9315554Publisher: Elsevier
Correspondence: nishiuchi@med.osaka-cu.ac.jp
- Article ID: 1468
Knirel YA, Kocharova NA, Bystrova OV, Katzenellenbogen E, Gamian A "Structures and serology of the O-specific polysaccharides of bacteria of the genus Citrobacter" -
Archivum Immunologiae et Therapiae Experimentalis 50(6) (2002) 379-391
The review presents the structures of the O-specific polysaccharides (O-antigens) of the lipopolysaccharides isolated from over 25 Citrobacter strains, which represent different species and serogroups. The correlation between O-antigen structure and immunospecificity as well as numerous cross-reactions between Citrobacter and other enterobacterial species are discussed.
Lipopolysaccharide, structure, O-antigen, O-specific polysaccharide, serology, Citrobacter, immunospecificity
NCBI PubMed ID: 12546064Journal NLM ID: 0114365Publisher: Basel, Boston: Birkhaüser
Correspondence: knirel@ioc.ac.ru
Institutions: N.D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia
- 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: 3313
Zhao G, Liu J, Liu X, Chen M, Zhang H, Wang PG "Cloning and characterization of GDP-perosamine synthetase (Per) from Escherichia coli O157:H7 and synthesis of GDP-perosamine in vitro" -
Biochemical and Biophysical Research Communications 363(3) (2007) 525-530
GDP-perosamine synthetase (Per, E.C. not yet classified) is important to the synthesis of Escherichia coli O157:H7 O-antigen. The mutant in per gene can disrupt the synthesis of O157 O-antigen. In this study, GDP-perosamine synthetase was cloned from E. coli O157:H7 and over-expressed in E. coli BL21 (DE3). The recombinant His-tagged Per fusion protein was a decamer with molecular weight of 431 kDa. The optimal pH value of this recombinant protein was 7.5. The divalent ions had no significant effect on Per-catalyzed reaction. The K(m) and K(cat)/K(m) for GDP-4-keto-6-deoxy-d-mannose were 0.09 mM and 2.1 x 10(5)M(-1)S(-1), and those for l-glutamate were 2mM and 0.52 x 10(5)M(-1)S(-1), respectively. Per was used to synthesize GDP-perosamine from GDP-mannose together with recombinant GDP-mannose dehydratase (GMD, E.C. 4.2.1.47). The purified GDP-perosamine was identified by MS and NMR. In summary, this work provided a feasible approach for the synthesis of GDP-perosamine which can lead to the study of LPS biosynthesis of pathogenic E. coli O157:H7
E. coli O157:H7, GDP-perosamine, GDP-perosamine synthetase, Per, Drug design
NCBI PubMed ID: 17888872Journal NLM ID: 0372516Publisher: Academic Press
Correspondence: zhanghoucheng@sdu.edu.cn
Institutions: The State Key Laboratory of Microbial Technology, Shandong University, Jinan, Shandong 250100, China
Methods: 13C NMR, 1H NMR, SDS-PAGE, ESI-MS, genetic methods, biochemical methods, capillary electrophoresis (CE)
- Article ID: 3847
Gajdus J, Glosnicka R, Szafranek J "Primary structure of Salmonella spp. O-antigens" -
Wiadomosci Chemiczne [Polish] 60(9-10) (2006) 621-653
Salmonella spp. are pathogenic Gram-negative bacteria that belong to Enterobacteriaceae family with lipopolysaccharide (LPS) as a constituent of cell wall. This is an integral component of the outer membrane of the wall. Salmonella smooth (S) forms produce LPS, which is composed of three parts, chemically bonded together viz. polysaccharide O-antigen, oligosaccharide core region and lipid A. Antigens O (O-PS) together with H flagella antigens are the foundation of serological classification of these bacteria. O-chain, which is built with up to 50 oligosaccharide repeating units, is one of the products of mild acidic hydrolysis of LPS. Due to the fact that polysaccharide antigens are the sites of specific antibody complexing, any difference in primary and secondary structures of O-antigens reflect serological specificity of bacteria. Taking this fact into consideration, we can distinguish about 2541 Salmonella serotypes with O and H antigenic formulas defined [4]. In this review we present 55 chemical structures of O-antigenic repeating units of Salmonella strains including their heterogeneity structures. The structures can have 22 different monosaccharide residues usually in 3 to 6 sugar repeating units. We describe here selected chemical and spectroscopic (MS, NMR) methods for primary structure examination of these bacterial O-PS. Enzymatic and immunochemical methods are also described. Cross-reactions of Salmonella spp. with any other bacteria or blood group A, B, 0 antigens are explained on the molecular level. Thus, structural assignments of somatic antigens of Salmonella spp. allow us to understand the molecular level of the classification system of these bacteria.
NMR spectroscopy, O-antigens, Salmonella, MS, primary structure
WWW link: http://baztech.icm.edu.pl/baztech/cgi-bin/btgetdoc.cgi?BUS2-0016-0014Publisher: Polish Chemical Society
Correspondence: jerzyg@chemik.chem.univ.gda.pl
Institutions: Wydzial Chemii, Uniwersytet Gdanski, ul. Sobieskiego 18, 80-952 Gdansk
- Article ID: 4300
Gao Y, Liu B, Strum S, Schutzbach JS, Druzhinina TN, Utkina NS, Torgov VI, Danilov LL, Veselovsky VV, Vlahakis JZ, Szarek WA, Wang L, Brockhausen I "Biochemical characterization of WbdN, a b1,3-glucosyltransferase involved in O-antigen synthesis in enterohemorrhagic Escherichia coli O157" -
Glycobiology 22(8) (2012) 1092-1102
The enterohemorrhagic O157 strain of Escherichia coli, which is one of the most well known bacterial pathogens, has an O-antigen repeating unit structure with the sequence [-2-D-Rha4NAcα1-3-L-Fucα1-4-D-Glcβ1-3-D-GalNAcα1-]. The O-antigen gene cluster of E. coli O157 contains the genes responsible for the assembly of this repeating unit and includes wbdN. In spite of cloning many O-antigen genes, biochemical characterization has been done on very few enzymes involved in O-antigen synthesis. In this work we expressed the wbdN gene in E. coli BL21, and the His-tagged protein was purified. WbdN activity was characterized using the donor substrate UDP-[(14)C]Glc and the synthetic acceptor substrate GalNAcα-O-PO(3)-PO(3)-(CH(2))(11)-O-Ph. The enzyme product was isolated by HPLC, and mass spectrometry showed that one Glc residue was transferred to the acceptor by WbdN. NMR analysis of the product structure indicated that Glc was β1-3 linked to GalNAc. WbdN contains a conserved DxD motif and requires divalent metal ions for full activity. WbdN activity has an optimal pH between 7 and 8, and is highly specific for UDP-Glc as the donor substrate. GalNAcα derivatives lacking the diphosphate group were inactive as substrates, and the enzyme did not transfer Glc to GlcNAcα-O-PO(3)-PO(3)-(CH(2))(11)-O-Ph. Our results illustrate that WbdN is a specific UDP-Glc: GalNAcα-diphosphate-lipid β1,3-Glc-transferase. The enzyme is a target for the development of inhibitors to block O157-antigen synthesis.
Escherichia coli O157, Substrate Specificity, glucosyltransferase, gastrointestinal infections, WbdN
NCBI PubMed ID: 22556057Publication DOI: 10.1093/glycob/cws081Journal NLM ID: 9104124Publisher: IRL Press at Oxford University Press
Correspondence: I. Brockhausen
Institutions: Department of Medicine, Division of Rheumatology, and Department of Biomedical and Molecular Sciences, Queen's University, Kingston, Ontario, Canada
Methods: 13C NMR, 1H NMR, NMR-2D, 31P NMR, ESI-MS, Western blotting, NMR-1D, biochemical methods, HPLC, enzymatic 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
- Article ID: 4404
Thoden JB, Reinhardt LA, Cook PD, Menden P, Cleland WW, Holden HM "The Catalytic Mechanism of Perosamine N-Acetyltransferase Revealed by High Resolution X-ray Crystallographic Studies and Kinetic Analyses" -
Biochemistry 51(16) (2012) 3433-3444
N-acetylperosamine is an unusual dideoxysugar found in the O-antigens of some Gram-negative bacteria including the pathogenic Escherichia coli strain O157:H7. The last step in its biosynthesis is catalyzed by PerB, an N-acetyltransferase belonging to the left-handed beta-helix superfamily of proteins. Here we describe a combined structural and functional investigation on PerB from Caulobacter crescentus. For this study, three structures were determined to 1.0 A resolution or better: the enzyme in complex with CoA and GDP-perosamine, the protein with bound CoA and GDP-N-acetylperosamine, and the enzyme containing a tetrahedral transition state mimic bound in the active site. Each subunit of the trimeric enzyme folds into two distinct regions. The N-terminal domain is globular and dominated by a six-stranded mainly parallel beta-sheet. It provides most of the interactions between the protein and GDP-perosamine. The C-terminal domain consists of a left-handed beta-helix, which has nearly seven turns. This region provides the scaffold for CoA binding. On the basis of these high-resolution structures, site-directed mutant proteins were constructed to test the roles of His 141 and Asp 142 in the catalytic mechanism. Kinetic data and pH rate profiles are indicative of His 141 serving as a general base. In addition, the backbone amide group of Gly 159 provides an oxyanion hole for stabilization of the tetrahedral transition state. The pH rate profiles are also consistent with the GDP-linked amino sugar substrate entering the active site in its unprotonated form. Finally, for this investigation, we show that PerB can accept GDP-3-deoxyperosamine as an alternative substrate, thus representing the production of a novel trideoxysugar.
biosynthesis, O-antigen, X-ray, Escherichia coli, gram negative bacteria, binding, Caulobacter crescentus, perosamine N-acetyltransferase, N-acetylperosamine
NCBI PubMed ID: 22443398Publication DOI: 10.1021/bi300197hJournal NLM ID: 0370623Publisher: American Chemical Society
Correspondence: Hazel_Holden@biochem.wisc.edu; cleland@biochem.wisc.edu
Institutions: Department of Biochemistry, University of Wisconsin, Madison, WI, USA
Methods: 1H NMR, X-ray, ESI-MS, biochemical methods, HPLC, crystallization
- Article ID: 4517
Gao Y, Vinnikova A, Brockhausen I "Functional Identification of Bacterial Glucosyltransferase WbdN" -
Book: Methods in Molecular Biology (2013) Vol. 1022, 199-214
The outer membrane of gram-negative bacteria is stabilized by lipopolysaccharides (LPS). The O-antigenic polysaccharides of LPS are composed of repeating units that are exposed to and can interact with the environment. The glycosyltransferases that assemble these repeating units are encoded by the O-antigen gene cluster and utilize undecaprenol-phosphate-linked intermediates as natural acceptor substrates, and nucleotide sugars as donor substrates on the cytoplasmic face of the inner membrane. Many of the glycosyltransferase genes are known but the enzymatic functions of most of them remain to be identified. We describe here how the function of a recombinant glucosyltransferase WbdN from Escherichia coli O157 can be determined by NMR analysis of the enzyme product, using a synthetic acceptor substrate analog. A fluorescent acceptor substrate analog can be used in highly sensitive enzyme assays that allow the characterization of enzyme activity without the use of radioactive nucleotide sugar donor substrates.
NMR, glucosyltransferase, linkage analysis, WbdN, fluorescent acceptor substrate
NCBI PubMed ID: 23765664Publication DOI: 10.1007/978-1-62703-465-4_16Publisher: Totowa, NJ: Humana Press
Correspondence: I. Brockhausen
Editors: Holst O, Walker JM, Beck A
Institutions: Department of Medicine and Department of Biomedical and Molecular Sciences, Queen's University, Kingston, Canada
Methods: 13C NMR, 1H NMR, NMR-2D, SDS-PAGE, sugar analysis, 31P NMR, MALDI-MS, Western blotting, NMR-1D, biochemical methods, HPLC
- Article ID: 5185
Martinez-Gomez E, Stahle J, Gil-Ramirez Y, Zuniga-Ripa A, Zaccheus M, Moriyón I, Iriarte M, Widmalm G, Conde-Alvarez R "Genomic Insertion of a Heterologous Acetyltransferase Generates a New Lipopolysaccharide Antigenic Structure in Brucella abortus and Brucella melitensis" -
Frontiers in Microbiology 9 (2018) 1092
Brucellosis is a bacterial zoonosis of worldwide distribution caused by bacteria of the genus Brucella. In Brucella abortus and Brucella melitensis, the major species infecting domestic ruminants, the smooth lipopolysaccharide (S-LPS) is a virulence factor. This S-LPS carries a N-formyl-perosamine homopolymer O-polysaccharide that is the major antigen in serodiagnostic tests and is required for virulence. We report that the Brucella O-PS can be structurally and antigenically modified using wbdR, the acetyl-transferase gene involved in N-acetyl-perosamine synthesis in Escherichia coli O157:H7. Brucella constructs carrying plasmidic wbdR expressed a modified O-polysaccharide but were unstable, a problem circumvented by inserting wbdR into a neutral site of chromosome II. As compared to wild-type bacteria, both kinds of wbdR constructs expressed shorter O-polysaccharides and NMR analyses showed that they contained both N-formyl and N-acetyl-perosamine. Moreover, deletion of the Brucella formyltransferase gene wbkC in wbdR constructs generated bacteria producing only N-acetyl-perosamine homopolymers, proving that wbdR can replace for wbkC. Absorption experiments with immune sera revealed that the wbdR constructs triggered antibodies to new immunogenic epitope(s) and the use of monoclonal antibodies proved that B. abortus and B. melitensis wbdR constructs respectively lacked the A or M epitopes, and the absence of the C epitope in both backgrounds. The wbdR constructs showed resistance to polycations similar to that of the wild-type strains but displayed increased sensitivity to normal serum similar to that of a per R mutant. In mice, the wbdR constructs produced chronic infections and triggered antibody responses that can be differentiated from those evoked by the wild-type strain in S-LPS ELISAs. These results open the possibilities of developing brucellosis vaccines that are both antigenically tagged and lack the diagnostic epitopes of virulent field strains, thereby solving the diagnostic interference created by current vaccines against Brucella.
antigen, Brucella, acetyltransferase, lipopolysaccharide (LPS), virulence factor, brucellosis, bacterial pathogenesis, vaccine development
NCBI PubMed ID: 29887851Publication DOI: 10.3389/fmicb.2018.01092Journal NLM ID: 101548977Publisher: Lausanne: Frontiers Research Foundation
Correspondence: Raquel Conde-Бlvarez
Institutions: Department of Organic Chemistry, Arrhenius Laboratory, Stockholm University, Stockholm, Sweden, Instituto de Salud Tropical, Instituto de Investigacion Sanitaria de Navarra, Departamento de Microbiologia y Parasitologia, Universidad de Navarra, Pamplona, Spain
Methods: 13C NMR, 1H NMR, NMR-2D, SDS-PAGE, DNA techniques, ELISA, mild acid hydrolysis, Western blotting, biological assays, serum bactericidal assays, immunological assays
- Article ID: 5423
Dobrochaeva KL, Khasbiulina NR, Shilova NV, Obukhova PS, Knirel YA, Nokel AY, Bovin NV "Human antibodies eluted from ligand-free Sepharose capable of binding bacterial polysaccharides and sulfated glycans" -
Molecular Immunology 106 (2019) 63-68
Sepharose matrix without immobilized ligands binds antibodies from human blood serum or immunoglobulin preparations. The eluted antibodies bind bacterial polysaccharides having no structural similarity to agarose (Sepharose is a cross-linked polysaccharide agarose) with a high affinity. It is concluded that the identified antibodies are capable of recognizing spatial rather than linear epitopes of bacterial polysaccharides. This side activity of Sepharose matrix should be taken into account in isolating target antibodies and other proteins from human blood.
antibodies, bacteria, polysaccharides, printed glycan array, Agarose, Sepharose
NCBI PubMed ID: 30583222Publication DOI: 10.1016/j.molimm.2018.12.011Journal NLM ID: 7905289Publisher: Elsevier
Correspondence: N.V. Bovin
Institutions: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 16/10 ul. Miklukho-Maklaya, Moscow, Russian Federation, Semiotik LLC, Moscow, Russian Federation, N. D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, 47 Leninsky pr., Moscow, Russian Federation, School of Engineering, Computer & Mathematical Sciences, Auckland University of Technology, Auckland 1010, New Zealand
Methods: serological methods, UV, affinity chromatography, antibody binding, glycan array analysis, microarray binding assays, serum analysis, isolation of antibodies
- Article ID: 5472
Liu B, Furevi A, Perepelov AV, Guo X, Cao H, Wang Q, Reeves PR, Knirel YA, Wang L, Widmalm G "Structure and genetics of Escherichia coli O antigens" -
FEMS Microbiology Reviews 44(6) (2020) 655-683
Escherichia coli includes clonal groups of both commensal and pathogenic strains, with some of the latter causing serious infectious diseases. O antigen variation is current standard in defining strains for taxonomy and epidemiology, providing the basis for many serotyping schemes for Gram-negative bacteria. This review covers the diversity in E. coli O antigen structures and gene clusters, and the genetic basis for the structural diversity. Of the 187 formally defined O antigens, six (O31, O47, O67, O72, O94 and O122) have since been removed and four (O14, O34, O89 and O144) strains do not produce any O antigen. Therefore, structures are presented for 176 of the 181 E. coli O antigens, some of which include subgroups. Most (93%) of these O antigens are synthesized via the Wzx/Wzy pathway, 11 via the ABC transporter pathway, with O20, O57 and O60 still uncharacterized due to failure to find their O antigen gene clusters. Biosynthetic pathways are given for 38 of the 49 sugars found in E. coli O antigens, and several pairs or groups of the E. coli antigens that have related structures show close relationships of the O antigen gene clusters within clades, thereby highlighting the genetic basis of the evolution of diversity.
structure, O antigen, Escherichia coli, gene cluster, serogroup, diversity
NCBI PubMed ID: 31778182Publication DOI: 10.1093/femsre/fuz028Journal NLM ID: 8902526Publisher: Oxford University Press
Correspondence: G. Widmalm
; Lei Wang
Institutions: Department of Organic Chemistry, Arrhenius Laboratory, Stockholm University, Stockholm, Sweden, N.D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia, Tianjin Key Laboratory of Microbial Functional Genomics, Tianjin, China, The Key Laboratory of Molecular Microbiology and Technology, Ministry of Education, Tianjin, China, School of Molecular and Microbial Bioscience (G08), University of Sydney, Sydney, Australia, TEDA Institute of Biological Sciences and Biotechnology, Nankai University, TEDA, Tianjin, China, Department of Immunology, School of Basic Medical Sciences, Tianjin Medical University, Tianjin, China
- Article ID: 5535
Plattner M, Shneider MM, Arbatsky NP, Shashkov AS, Chizhov AO, Nazarov S, Taylor NMI, Prokhorov NS, Buth SA, Gambino M, Gencay YE, Brondsted L, Kutter EM, Knirel YA, Leiman PG "Structure and function of the branched receptor-binding complex of bacteriophage CBA120" -
Journal of Molecular Biology 431 (2019) 3718-3719
Bacteriophages recognize their host cells with the help of tail fiber and tailspike proteins that bind, cleave, or modify certain structures on the cell surface. The spectrum of ligands to which the tail fibers and tailspikes can bind is the primary determinant of the host range. Bacteriophages with multiple tailspike/tail fibers are thought to have a wider host range than their less endowed relatives but the function of these proteins remains poorly understood. Here, we describe the structure, function, and substrate specificity of three tailspike proteins of bacteriophage CBA120-TSP2, TSP3 and TSP4 (orf211 through orf213, respectively). We show that tailspikes TSP2, TSP3 and TSP4 are hydrolases that digest the O157, O77, and O78 Escherichia coli O-antigens, respectively. We demonstrate that recognition of the E. coli O157:H7 host by CBA120 involves binding to and digesting the O157 O-antigen by TSP2. We report the crystal structure of TSP2 in complex with a repeating unit of the O157 O-antigen. We demonstrate that according to the specificity of its tailspikes TSP2, TSP3, and TSP4, CBA120 can infect E. coli O157, O77, and O78, respectively. We also show that CBA120 infects Salmonella enterica serovar Minnesota, and this host range expansion is likely due to the function of TSP1. Finally, we describe the assembly pathway and the architecture of the TSP1-TSP2-TSP3-TSP4 branched complex in CBA120 and its related ViI-like phages.
NMR, X-ray crystallography, bacterial surface polysaccharides, bacteriophage attachment, host-cell recognition
NCBI PubMed ID: 31325442Publication DOI: 10.1016/j.jmb.2019.07.022Journal NLM ID: 2985088RPublisher: Elsevier
Correspondence: Petr G. Leiman
Institutions: N. D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia, Department of Biochemistry and Molecular Biology, Sealy Center for Structural Biology and Molecular Biophysics, University of Texas Medical Branch, 301 University Blvd, Galveston, TX 77555-0647, USA, Ecole Polytechnique Federale de Lausanne, Lausanne CH-1015, Switzerland, Laboratory of Molecular Bioengineering, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, 16/10 Miklukho-Maklaya St., 117997 Moscow, Russia, Structural Biology of Molecular Machines Group, Protein Structure & Function Programme, Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, Blegdamsvej 3B, Copenhagen 2200, Denmark, Department of Veterinary and Animal Sciences, University of Copenhagen, Stigbojlen 4, 1870 Frederiksberg C, Denmark, The Evergreen State College, Olympia, WA 98505, USA
Methods: 13C NMR, 1H NMR, NMR-2D, X-ray, SDS-PAGE, electron microscopy, cloning, bioinformatic analysis, crystallization, binding assays, HR-ESI-MS, bacteriophage digestion, phage characterization
- Article ID: 5742
Caroff M, Novikov A "Lipopolysaccharides: structure, function and bacterial identification" -
OCL - Oilseeds and fats, Crops and Lipids 27 (2020) 31
Lipopolysaccharides (LPS) are the main components of the outer membrane of Gram-negative bacteria. They are glycolipids containing a lipid moiety called lipid A, more often made of a bis-phosphorylated glucosamine disaccharide, carrying fatty acids in ester and amide linkages. Lipid A is linked to a core oligosaccharide of about 10 sugars, substituted in the wild-type strains, by long-chain oligosaccharide repetitive units, extending outside the bacteria and representing their main antigens. In addition to determine the serotype of the bacterium, LPS are highly potent biological molecules, capable of eliciting at the level of minute amounts, beneficial, as well as deleterious activities.
Lipopolysaccharide, serology, endotoxins, inflammation, structure-activity
Publication DOI: 10.1051/ocl/2020025Publisher: France: EDP Sciences (ISSN: 22726977, 22576614)
Correspondence: martine.caroff@lpsbiosciences.com
Institutions: LPS-BioSciences, Paris-Saclay University, Orsay, France, Hephaistos-Pharma, Paris-Saclay University, Orsay, France
- 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: 409
Structure type: homopolymer
; n=100
Compound class: O-polysaccharide
Contained glycoepitopes: IEDB_131172,IEDB_134281,IEDB_1397515,IEDB_2116320,IEDB_434547,IEDB_628715
The structure is contained in the following publication(s):
- Article ID: 138
Ugalde JE, Czibener C, Feldman MF, Ugalde RA "Identification and characterization of the Brucella abortus phosphoglucomutase gene: role of lipopolysaccharide in virulence and intracellular multiplication" -
Infection and Immunity 68(10) (2000) 5716-5723
Smooth lipopolysaccharide (LPS) of Brucella abortus has been reported to be an important virulence factor, although its precise role in pathogenesis is not yet clear. While the protective properties of LPS against complement are well accepted, there is still some controversy about the capacity of rough mutants to replicate intracellularly. The B. abortus phosphoglucomutase gene (pgm) was cloned, sequenced, and disrupted. The gene has a high index of identity to Agrobacterium tumefaciens pgm but is not part of the glycogen operon. A B. abortus null mutant lacks LPS O antigen but has an LPS core with an electrophoretic profile undistinguishable from that of the wild-type core, suggesting that glucose, galactose, or a derivative of these sugars may be part of the linkage between the core and the O antigen. This mutant is unable to survive in mice but replicates in HeLa cells, indicating that the complete LPS is not essential either for invasion or for intracellular multiplication. This behavior suggests that the LPS may play a role in extracellular survival in the animal, probably protecting the cell against complement-mediated lysis, but is not involved in intracellular survival.
Lipopolysaccharide, gene, role, virulence, characterization, identification, Brucella, Brucella abortus, intracellular, multiplication, phosphoglucomutase
NCBI PubMed ID: 10992476Journal NLM ID: 0246127Publisher: American Society for Microbiology
Correspondence: rugalde@inti.gov.ar
Institutions: Instituto de Investigations Biotecnologicas-Instituto Tecnologico de Chascomus, Universidad National de General San Martin, Buenos Aires, Centra de Virologia Animal, CEVAN, Capital Federal, Argentina.
Methods: genetic methods
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4. Compound ID: 437
Structure type: homopolymer
; n=12-18
Compound class: O-polysaccharide
The structure is contained in the following publication(s):
- Article ID: 163
Villeneuve S, Boutonnier A, Mulard LA, Fournier JM "Immunochemical characterization of an Ogawa-Inaba common antigenic determinant of Vibrio cholerae O1" -
Microbiology 145(9) (1999) 2477-2484
Cholera remains an important public health problem in many parts of the world and the availability of an effective cholera vaccine is important for the prevention of cholera in the countries affected by this disease. Despite the appearance in 1992 of a new serogroup, 0139, of Vibrio cholerae, most of the cholera outbreaks are still caused by V. cholerae O1 biotype El Tor. Vaccine trials in Asia from 1968 to 1971, and studies of the production of serotype-specific antiserum in rabbits and of the protective activity of monoclonal antibodies against diarrhoeal disease in neonatal mice, have led to the conclusion that the Ogawa serotype contains a specific antigenic determinant whereas the Inaba serotype contains a different antigenic determinant that cross-reacts with the Ogawa serotype. By studying the binding of anti-Ogawa monoclonal antibodies to synthetic oligosaccharide fragments mimicking the Ogawa O-specific polysaccharide, it has been shown that the terminal monosaccharide, bearing the 2-O-methyl group in the O-specific polysaccharide, is most probably the serotype-specific determinant for the Ogawa strain. However, study of the binding of a monoclonal antibody recognizing both Ogawa and Inaba serotypes suggested partial recognition of the core as well as of the O-specific polysaccharide of the LPS of V. cholerae O1. To further characterize this antigenic determinant that is common to the Ogawa and Inaba serotypes, the core and the O-specific polysaccharide linked to the core of V. cholerae O1 LPS were purified by preparative electrophoresis. The O-specific polysaccharide linked to the core was subjected to periodate oxidation to destroy sugars from the core. Binding studies of these purified saccharide fragments to a monoclonal antibody which is protective in mice and specific to the antigenic determinant common to Ogawa and Inaba serotypes showed that both the core and the O-specific polysaccharide are involved in this common antigenic determinant. This explains how the presence or the absence of the Ogawa-specific antigenic determinant would lead to the expression of two independent antigenic determinants of V. cholerae O1, one specific to the Ogawa serotype and the other common to both Ogawa and Inaba serotypes.
LPS, common, core, characterization, antigenic determinant, immunochemical, Vibrio, antigenic, determinant, Vibrio cholerae, Vibrio cholerae O1, preparative electrophoresis
NCBI PubMed ID: 10517600Journal NLM ID: 0376646Publisher: Washington, DC: Kluwer Academic/Plenum Publishers
Correspondence: fournier@pasteur.fr
Institutions: Unite du Cholera et des Vibrions, Centre National de Reference des Vibrions et du Cholera, Unite de Chimie Organique, Institut Pasteur, 25 rue du Dr. Roux, 75724 Paris Cedex 15, France.
Methods: serological methods
- Article ID: 1452
Chatterjee SN, Chaudhuri K "Lipopolysaccharides of Vibrio cholerae. I. Physical and chemical characterization" -
Biochimica et Biophysica Acta 1639(2) (2003) 65-79
Vibrio cholerae is the causative organism of the disease cholera. The lipopolysaccharide (LPS) of V. cholerae plays an important role in eliciting the antibacterial immune response of the host and in classifying the vibrios into some 200 or more serogroups. This review presents an account of our up-to-date knowledge of the physical and chemical characteristics of the three constituents, lipid-A, core-polysaccharide (core-PS) and O-antigen polysaccharide (O-PS), of the LPS of V. cholerae of different serogroups including the disease-causing ones, O1 and O139. The structure and occurrence of the capsular polysaccharide (CPS) on V. cholerae O139 have been discussed as a relevant topic. Similarity and dissimilarity between the structures of LPS of different serogroups, and particularly between O22 and O139, have been analysed with a view to learning their role in the causation of the epidemic form of the disease by avoiding the host defence mechanism and in the evolution of the newer pathogenic strains in future. An idea of the emerging trends of research involving the use of immunogens prepared from synthetic oligosaccharides that mimic terminal epitopes of the O-PS of V. cholerae O1 in the development of a conjugate anti cholera vaccine is also discussed.
Lipopolysaccharide, structure, lipid A, capsular polysaccharide, serogroup, Vibrio cholerae
NCBI PubMed ID: 14559113Publication DOI: 10.1016/j.bbadis.2003.08.004Journal NLM ID: 0217513Publisher: Elsevier
Correspondence: sncac@sify.com (S.N. Chatterjee)
Institutions: Saha Institute of Nuclear Physics, 1/AF Bidhannagar, Sector-1, Calcutta-700 064, India, Biophysics Division, Indian Institute of Chemical Biology, Jadavpur, Calcutta-700 032, India
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5. Compound ID: 438
Structure type: monomer
Trivial name: methyl 4,6-dideoxy-4-(3-deoxy-L-glycero-tetronamido)-α-D-mannopyranoside
The structure is contained in the following publication(s):
- Article ID: 164
Villeneuve S, Souchon H, Riottot M, Mazie J, Lei PS, Glaudemans CPJ, Kovác P, Fournier JM, Alzari PM "Crystal structure of an anti-carbohydrate antibody directed against Vibrio cholerae O1 in complex with antigen: Molecular basis for serotype specificity" -
Proceedings of the National Academy of Sciences of the USA 97(15) (2000) 8433-8438
The crystal structure of the murine Fab S-20-4 from a protective anti-cholera Ab specific for the lipopolysaccharide Ag of the Ogawa serotype has been determined in its unliganded form and in complex with synthetic fragments of the Ogawa O-specific polysaccharide (O-SP). The upstream terminal O-SP monosaccha-ride is shown to be the primary antigenic determinant. Additional perosamine residues protrude outwards from the Ab surface and contribute only marginally to the binding affinity and specificity. A complementary water-excluding hydrophobic interface and five Ab-Ag hydrogen bonds are crucial for carbohydrate recognition. The structure reported here explains the serotype specificity of anti-Ogawa Abs and provides a rational basis toward the development of a synthetic carbohydrate-based anti-cholera vaccine.
antigen, lipopolysaccharides, LPS, structure, serotype, antigenic determinant, molecular, antibodies, antibody, complex, specificity, crystal, crystal structure, Vibrio, antigenic, determinant, Vibrio cholerae, Vibrio cholerae O1, anti-carbohydrate, Serotypes
NCBI PubMed ID: 10880560Journal NLM ID: 7505876Publisher: National Academy of Sciences
Correspondence: alzarie@pasteur.fr
Institutions: Unite de Biochimie Structurale (Centre National de la Recherche Scientifique, Unite de Recherche Associee 2185), Unite du Cholera et des Vibrions, Unite d'lmmunologie Structurale, and 'Laboratoire d'lngenierie des Anticorps, Institut Pasteur, 25 rue du Dr. Roux, 75724 Paris, France, Laboratory of Medicinal Chemistry, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892-0815.
Methods: serological methods
- Article ID: 352
Poirot E, Zhang X, Whittaker NF, Kovác P "Syntheses of the L-manno and some other analogs of the terminal determinants of the O-PS of Vibrio cholerae O:1" -
Carbohydrate Research 330(1) (2001) 7-20
Analogs of the methyl α-glycosides of the terminal residues of the O-specific polysaccharides (O-PS) of Vibrio cholerae O:1, serotype Inaba and Ogawa, have been prepared as probes to study their interaction with anti V. cholerae O:1 antibodies. They differ from the termini of the respective O-PSs in anomeric or absolute configuration of perosamine, position of the O-methyl group in D-perosamine, and nature of the N-acyl side chain.
terminal, O-polysaccharide, Vibrio, determinant, Vibrio cholerae
NCBI PubMed ID: 11217964Journal NLM ID: 0043535Publisher: Elsevier
Correspondence: kpn@helix.nih.gov
Institutions: National Institutes of Health, Building 8, Room B1A24, Bethesda, MD 20892-0815, USA
Methods: chemical methods
- Article ID: 413
Wang J, Villeneuve S, Zhang J, Lei PS, Miller CE, Lafaye P, Nato F, Szu SSC, Karpas A, Bystricky S, Robbins JB, Kovác P, Fournier JM, Glaudemans CPJ "On the antigenic determinants of the lipopolysaccharides of Vibrio cholerae O:1, serotypes Ogawa and Inaba" -
Journal of Biological Chemistry 273(5) (1998) 2777-2783
Monoclonal, murine IgG1s S-20-4, A-20-6, and IgA 2D6, directed against Vibrio cholerae O:1 Ogawa-lipopolysaccharide exhibited the same fine specificities and similar affinities for the synthetic methyl α-glycosides of the (oligo)saccharide fragments mimicking the Ogawa O-polysaccharide (O-PS). They did not react with the corresponding synthetic fragments of Inaba O-PS. IgG1s S-20-4 and A-20-6 have absolute affinity constants for synthetic Ogawa mono- to hexasaccharides of from approximately 10(5) to approximately 10(6) M-1. For IgG1s S-20-4, A-20-6, and IgA 2D6, the nonreducing terminal residue of Ogawa O-PS is the dominant determinant, accounting for approximately 90% of the maximal binding energy shown by these antibodies. Binding studies of derivatives of the Ogawa monosaccharide and IgGs S-20-4 and A-20-6 revealed that the C-2 O-methyl group fits into a somewhat flexible antibody cavity and that hydrogen bonds involving the oxygen and, respectively, the OH at the 2- and 3-position of the sugar moiety as well as the 2'-position in the amide side chain are required. Monoclonal IgA ZAC-3 and IgG3 I-24-2 are specific for V. cholerae O:1 serotypes Ogawa/Inaba-LPS.1 The former did not show binding with members of either series of the synthetic ligands related to the O-antigens of the Ogawa or Inaba serotypes, in agreement with its reported specificity for the lipid/core region (1). Inhibition studies revealed that the binding of purified IgG3 I-24-2 to Ogawa-LPS might be mediated by a region in the junction of the OPS to the lipid-core region of the LPS. cDNA cloning and analysis of the anti-Ogawa antibodies S-20-4, A-20-6, and 2D6 revealed a very high degree of homology among the heavy chains. Among the light chains, no such homology between S-20-4 and A-20-6 on the one hand, and 2D6 on the other hand, exists. For the anti-Inaba/Ogawa antibodies I-24-2 and ZAC-3, their heavy chains are completely different, with some homology among the light chains
Lipopolysaccharide, lipopolysaccharides, LPS, serotype, antigenic determinant, Vibrio, antigenic, determinant, Vibrio cholerae, Serotypes
NCBI PubMed ID: 9446585Journal NLM ID: 2985121RPublisher: Baltimore, MD: American Society for Biochemistry and Molecular Biology
Correspondence: glau@helix.nih.gov
Institutions: Laboratory of Medicinal Chemistry, NIDDK and the Laboratory of Developmental and Molecular Immunity, NICHD, National Institutes of Health, Bethesda, MD, USA, Unite du Cholera et des Vibrions, Centre National de Reference des Vibrions et du Cholera and Hybridolab, Institut Pasteur, Paris, France
Methods: 1H NMR, PCR, DNA sequencing, SDS-PAGE, ELISA, genetic methods
- Article ID: 924
Lei PS, Ogawa Y, Flippen-Anderson JL, Kovác P "Synthesis and crystal structure of methyl 4,6-dideoxy-4-(3-deoxy-L-glycero-tetronamido)-2-O-methyl-a-D-mannopyranoside, the methyl a-glycoside of the terminal unit, and presumed antigenic determinant, of the O-specific polysaccharide of Vibrio cholerae O:1, serotype Ogawa" -
Carbohydrate Research 275(1) (1995) 117-129
Methyl 4-azido-4,6-dideoxy-3-O-benzyl-α-D-mannopyranoside and its analogous 3-O-(4-methoxybenzyl) derivative were methylated and the 2-O-methyl derivatives formed were convetted into methyl 4-amino-4,6-dideoxy-2-O-methyl-α-o-mannopyranoside. Reaction of the latter with 3-deoxy-L-glycero-tetronolactone gave the methyl glycoside of 4,6-dideoxy-4-(3-deoxy-L-glycero-tetronamido)-2-O-methyl-α-D-mannopyranose, the monosaccharide that is reported to be the terminal moiety of the O-specific polysaccharide of Vibrio cholerae O:1, serotype Ogawa. The unit cell packing of the compound, which crystallized as a monohydrate, differs from that of the previously described crystalline compound lacking the 2-O-methyl group. The unmethylated sugar is the terminal moiety of the O-specific polysaccharide of Vibrio cholerae O:1, serotype Inaba. The crystal structure of methyl 4,6-dideoxy-2-O-methyl-4-trifluoroacetamido-α-D-mannopyranoside is also described.
synthesis, structure, terminal, polysaccharide, serotype, D-mannose, antigenic determinant, O-polysaccharide, O-specific, O-specific polysaccharide, crystal, crystal structure, Vibrio, antigenic, determinant, Vibrio cholerae, Vibrio cholerae O1, glycoside, O-methyl, methyl
NCBI PubMed ID: 7585718Journal NLM ID: 0043535Publisher: Elsevier
Institutions: NIDDK, National Institutes of Health, Bethesda, MD, USA, Laboratory for the Structure of Matter, Naval Research Laboratory, Washington, DC, USA
- Article ID: 1396
Bystricky S, Szu SC, Gotoh M, Kovác P "Circular dichroism of the O-specific polysaccharide of Vibrio cholerae O1 and some related derivatives" -
Carbohydrate Research 270(2) (1995) 115-122
The O-specific polysaccharide (O-SP) of Vibrio cholerae O1 is a homopolymer of α-(1→2)-linked 4-amino-4, 6-dideoxy-D-mannopyranose whose amino group is acylated with 3-deoxy-L-glycero-tetronic acid [N-(3-deoxy-L-glycero- tetronyl)-α-D-perosamine]. The circular dichroism (CD) of the O-SP as well as of a number of N-acyl (formyl, acetyl, 4-hydroxybutyl, 3-deoxy-L-and D-glycero-tetronyl) derivatives of methyl α-glycosides of 4-amino-4,6-dideoxy-D-mannopyranose (methyl α-D-perosaminide) has been studied for solutions in water, acetonitrile and 1,1,1-trifluoroethanol. The strong solvent dependence of the sign and intensity of the CD observed for the monosaccharide amides bearing achiral acyl groups is explained by solvent-mediated change of the orientation of the amido group relative to the proximal hydroxyl group at C-3. A change in the population of the nonplanar conformers with a pyramidal arrangement of bonds at the amido nitrogen has also been considered. The effect of solvents upon the CD spectra of compounds bearing chiral N-acyl substituents is less pronounced than that of their counterparts bearing achiral N-acyl substituents. The sign of the CD for the O-SP was found negative in all solvents used. This result is in agreement with the negative sign of the CD of the n → pi electron transition observed, independent of the solvent, for the monosaccharide derivative containing the L-glycero-3-deoxytetronamido group, and the positive sign found for its D-glycero-counterpart.
polysaccharide, O-specific, O-specific polysaccharide, derivative, Vibrio, Vibrio cholerae, Vibrio cholerae O1, circular dichroism, Vibrio cholerae O1NT:
NCBI PubMed ID: 7585695Journal NLM ID: 0043535Publisher: Elsevier
Institutions: NICHD, Laboratory of Developmental and Molecular Immunity, National Institutes of Health, Bethesda, USA, NICHD, Laboratory of Developmental and Molekular Immunity, National Institutes of Health, Bethesda, USA
Methods: CD
- Article ID: 3294
Gotoh M, Barnes CN, Kovác P "Improved synthesis and the crystal structure of methyl 4,6-dideoxy-4-(3-deoxy-L-glycero- tetronamido)-a-D-mannopyranoside, the methyl a-glycoside of the intracatenary repeating unit of the O-polysaccharide of Vibrio cholerae O:1" -
Carbohydrate Research 260(2) (1994) 203-218
The crude product of deamination of the commercially available L-homoserine was acetylated and the 2-O-acetyl-3-deoxy-L-glycero-tetronolactone (18) formed was used to N-acylate methyl perosaminide (methyl 4-amino-4,6-dideoxy-α-D-mannopyranoside, 12) and its 2,3-O-isopropylidene derivative. The major product isolated from the reaction was the crystalline methyl 4-(4-O-acetyl-3-deoxy-L-glycero-tetronamido)-4,6-dideoxy-α-D-mannopyranoside (1, 70-75%) resulting from acetyl group migration in the initially formed 2'-O-acetyl derivative. O-Deacetylation of 1 gave the title amide 2. Compound 2, obtained crystalline for the first time, was fully characterized, and its crystal structure was determined. Deoxytetronamido derivatives diastereomeric with 1 and 2, respectively, were obtained by the acylation of 12 with 2-O-acetyl-3-deoxy-D-glycero-tetronolactone (prepared from D-homoserine), and subsequent deacetylation. Structures of several byproducts of the reaction of 12 with 18 have been deduced from their spectral characteristics. Since these byproducts were various O-acetyl derivatives of 2, the title compound could be obtained in approximately 90% yield by deacetylating (Zemplen) the crude mixture of N-acylation products, followed by chromatography.
synthesis, structure, polysaccharide, repeating unit, group, crystalline, O-polysaccharide, O polysaccharide, crystal, crystal structure, derivative, 2, Vibrio, Vibrio cholerae, chromatography, acetylated, monosaccharide, reaction, 4-amino-4, 6-dideoxy-D-mannose, PDF, O-acetyl, acylation, glycoside, methyl, characteristics, amide, deacetylation, 4-dihydroxybutyryl, Health
NCBI PubMed ID: 7520833Journal NLM ID: 0043535Publisher: Elsevier
Institutions: NIDDK, National Institutes of Health, Bethesda, MD, USA
Methods: 13C NMR, 1H NMR, chemical synthesis, crystallography
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6. Compound ID: 536
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S-2,4HOBut-(1-4)-+
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S-2,4HOBut-(1-4)-a-D-Rhap4N-(1-2)-a-D-Rhap4N-(1-1)-Me |
Show graphically |
Structure type: oligomer
The structure is contained in the following publication(s):
- Article ID: 164
Villeneuve S, Souchon H, Riottot M, Mazie J, Lei PS, Glaudemans CPJ, Kovác P, Fournier JM, Alzari PM "Crystal structure of an anti-carbohydrate antibody directed against Vibrio cholerae O1 in complex with antigen: Molecular basis for serotype specificity" -
Proceedings of the National Academy of Sciences of the USA 97(15) (2000) 8433-8438
The crystal structure of the murine Fab S-20-4 from a protective anti-cholera Ab specific for the lipopolysaccharide Ag of the Ogawa serotype has been determined in its unliganded form and in complex with synthetic fragments of the Ogawa O-specific polysaccharide (O-SP). The upstream terminal O-SP monosaccha-ride is shown to be the primary antigenic determinant. Additional perosamine residues protrude outwards from the Ab surface and contribute only marginally to the binding affinity and specificity. A complementary water-excluding hydrophobic interface and five Ab-Ag hydrogen bonds are crucial for carbohydrate recognition. The structure reported here explains the serotype specificity of anti-Ogawa Abs and provides a rational basis toward the development of a synthetic carbohydrate-based anti-cholera vaccine.
antigen, lipopolysaccharides, LPS, structure, serotype, antigenic determinant, molecular, antibodies, antibody, complex, specificity, crystal, crystal structure, Vibrio, antigenic, determinant, Vibrio cholerae, Vibrio cholerae O1, anti-carbohydrate, Serotypes
NCBI PubMed ID: 10880560Journal NLM ID: 7505876Publisher: National Academy of Sciences
Correspondence: alzarie@pasteur.fr
Institutions: Unite de Biochimie Structurale (Centre National de la Recherche Scientifique, Unite de Recherche Associee 2185), Unite du Cholera et des Vibrions, Unite d'lmmunologie Structurale, and 'Laboratoire d'lngenierie des Anticorps, Institut Pasteur, 25 rue du Dr. Roux, 75724 Paris, France, Laboratory of Medicinal Chemistry, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892-0815.
Methods: serological methods
- Article ID: 413
Wang J, Villeneuve S, Zhang J, Lei PS, Miller CE, Lafaye P, Nato F, Szu SSC, Karpas A, Bystricky S, Robbins JB, Kovác P, Fournier JM, Glaudemans CPJ "On the antigenic determinants of the lipopolysaccharides of Vibrio cholerae O:1, serotypes Ogawa and Inaba" -
Journal of Biological Chemistry 273(5) (1998) 2777-2783
Monoclonal, murine IgG1s S-20-4, A-20-6, and IgA 2D6, directed against Vibrio cholerae O:1 Ogawa-lipopolysaccharide exhibited the same fine specificities and similar affinities for the synthetic methyl α-glycosides of the (oligo)saccharide fragments mimicking the Ogawa O-polysaccharide (O-PS). They did not react with the corresponding synthetic fragments of Inaba O-PS. IgG1s S-20-4 and A-20-6 have absolute affinity constants for synthetic Ogawa mono- to hexasaccharides of from approximately 10(5) to approximately 10(6) M-1. For IgG1s S-20-4, A-20-6, and IgA 2D6, the nonreducing terminal residue of Ogawa O-PS is the dominant determinant, accounting for approximately 90% of the maximal binding energy shown by these antibodies. Binding studies of derivatives of the Ogawa monosaccharide and IgGs S-20-4 and A-20-6 revealed that the C-2 O-methyl group fits into a somewhat flexible antibody cavity and that hydrogen bonds involving the oxygen and, respectively, the OH at the 2- and 3-position of the sugar moiety as well as the 2'-position in the amide side chain are required. Monoclonal IgA ZAC-3 and IgG3 I-24-2 are specific for V. cholerae O:1 serotypes Ogawa/Inaba-LPS.1 The former did not show binding with members of either series of the synthetic ligands related to the O-antigens of the Ogawa or Inaba serotypes, in agreement with its reported specificity for the lipid/core region (1). Inhibition studies revealed that the binding of purified IgG3 I-24-2 to Ogawa-LPS might be mediated by a region in the junction of the OPS to the lipid-core region of the LPS. cDNA cloning and analysis of the anti-Ogawa antibodies S-20-4, A-20-6, and 2D6 revealed a very high degree of homology among the heavy chains. Among the light chains, no such homology between S-20-4 and A-20-6 on the one hand, and 2D6 on the other hand, exists. For the anti-Inaba/Ogawa antibodies I-24-2 and ZAC-3, their heavy chains are completely different, with some homology among the light chains
Lipopolysaccharide, lipopolysaccharides, LPS, serotype, antigenic determinant, Vibrio, antigenic, determinant, Vibrio cholerae, Serotypes
NCBI PubMed ID: 9446585Journal NLM ID: 2985121RPublisher: Baltimore, MD: American Society for Biochemistry and Molecular Biology
Correspondence: glau@helix.nih.gov
Institutions: Laboratory of Medicinal Chemistry, NIDDK and the Laboratory of Developmental and Molecular Immunity, NICHD, National Institutes of Health, Bethesda, MD, USA, Unite du Cholera et des Vibrions, Centre National de Reference des Vibrions et du Cholera and Hybridolab, Institut Pasteur, Paris, France
Methods: 1H NMR, PCR, DNA sequencing, SDS-PAGE, ELISA, genetic methods
- Article ID: 1397
Bystricky S, Szu SC, Zhang J, Kovác P "Conformational differences among mono- and oligosaccharide fragments of the O-specific polysaccharides of Vibrio cholerae O1 revealed by circular dichroism" -
Carbohydrate Research 314(1-2) (1998) 135-139
The circular dichroism (CD) of synthetic mono- and oligosaccharides that represent the terminal, non-reducing group of O-antigens of Vibrio cholerae O1 from the subtypes Ogawa and Inaba was measured in various solvents. We found differences in the CD of the monosaccharides of these subtypes that decrease with increasing chain lengths of the oligosaccharides. The differences can be explained by different orientations of the N-acyl side chain of the terminal monosaccharides. The linear relationship of ellipticity versus the number of residues in an oligosaccharide chain follows the principle of optical superposition. This, together with a similar contribution by internal units to the overall ellipticity, suggests an identical, regular conformation of oligosaccharide fragments of both Ogawa and Inaba series.
oligosaccharide, polysaccharide, conformational, O-antigens, O-specific, O-specific polysaccharide, polysaccharides, difference, fragment, Vibrio, Vibrio cholerae, Vibrio cholerae O1, O-specific polysaccharides, circular dichroism
NCBI PubMed ID: 10230041Journal NLM ID: 0043535Publisher: Elsevier
Institutions: Institute of Chemistry, Slovak Academy of Sciences, Bratislava, Slovak Republic
Methods: CD
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7. Compound ID: 866
Structure type: homopolymer
Trivial name: perosamine homopolymer, perosamine
Compound class: O-polysaccharide, O-antigen
Contained glycoepitopes: IEDB_131172,IEDB_134281,IEDB_1397515,IEDB_2116320,IEDB_434547,IEDB_628715
The structure is contained in the following publication(s):
- Article ID: 241
Godfroid F, Cloeckaert A, Taminiau B, Danese I, Tibor A, De Bolle X, Mertens P, Letesson JJ "Genetic organisation of the lipopolysaccharide O-antigen biosynthesis region of Brucella melitensis 16M (wbk)" -
Research in Microbiology 151(8) (2000) 655-668
Brucella spp. are Gram-negative, facultative intracellular bacteria that cause a zoonotic world-wide disease. As in other Gram-negative bacteria, its S-LPS (smooth lipopolysaccharide) is a major determinant of virulence. The Brucella melitensis 16M LPS O-antigen is a homopolymer of 4-formamido-4,6, dideoxymannose. In this study, the previously cloned 14-kb wbk gene cluster was sequenced, and seven open reading frames (ORFs) as well as four insertion sequences were identified. Six of the seven ORFs are homologous to LPS biosynthesis genes from other organisms. The gmd, per and wbkC gene products are predicted to be involved in 4-formamido-4,6,dideoxymannose synthesis. By deletion experiments, we demonstrated that the putative formyltransferase WbkC is absolutely required for the O-side-chain production. The wbkA gene product is similar to several mannosyltransferases and is probably involved in the polymerisation of the B. melitensis O-side-chain. We also identified two genes (wzm and wzt) encoding proteins with high similarity to several two-component ABC (ATP-binding cassette) transporters. Their implication in O-antigen translocation across the inner membrane was confirmed by gene replacement. Finally, no function has been assigned to the wbkB gene either by homology search or functionally, because deletion of wbkB did not interfere with the O-antigen structure. The seven ORFs have a low G + C content, indicating that they might have been acquired by lateral transfer from a progenitor with more A + T rich DNA
Lipopolysaccharide, biosynthesis, genetic, O-antigen, O antigen, O-side-chain, region, Brucella, ABC transporter, Brucella melitensis, lipopolysaccharide O-antigen, LPS genetics
NCBI PubMed ID: 11081580Journal NLM ID: 8907468Publisher: Elsevier
Correspondence: Jean-jaques.letesson@fundp.ac.be
Institutions: Unite de recherche en biologie moleculaire (URBM), Laboratoire d'immunologie et de microbiologie, Facultes universitaires Notre Dame de la Paix, Belgium
- Article ID: 269
Isshiki Y, Haishima Y, Kondo S, Hisatsune K "Immunochemistry of group A and Inaba C antigen factors constituting the O antigen of O1 Vibrio cholerae" -
European Journal of Biochemistry 229 (1995) 583-588
Serological cross-reactivity among intact lipopolysaccharides (LPS) from O1 Vibrio cholerae Inaba O-form (Inaba), Yersinia enterocolitica O9 (O9), non-O1 V. cholerae serogroup Hakata (Hakata) and Vibrio bio-serogroup 1875 Variant (1875 Variant) (all of which share Inaba antigen factor C), as well as a total of six kinds of chemically modified LPS (three from O9 and three from Inaba) was demonstrated by passive hemolysis and passive hemolysis inhibition by using these LPS as antigen for sensitizing sheep red blood cells and as inhibitor. These intact as well as chemically modified LPS contained, in their O polysaccharide chain, α(1→2)-linked linear perosamine (4-amino-4,6-dideoxy-D-manno-pyranose) homopolymers with different N-acyl groups: their acyl groups comprise 3-deoxy-L-glycero-tetronyl (Inaba LPS), formyl (O9 LPS), 3-hydroxypropionyl (1875 Variant LPS), acetyl (Hakata LPS and artificially introduced into Inaba and O9 LPS), propionyl and butyryl (both artificially introduced into Inaba and O9 LPS) groups. N-Deacylation of the α(l→2)-linked N-(3-deoxy-L-glycero-tetronyl)perosamine homopolymer of Inaba and the N-formyl one of O9 LPS resulted in virtual elimination of their serological reactivity with both homologous and heterologous antisera. Furthermore, when the resultant NH2 groups of the N-deacylated perosamine homopolymers of both LPS were N-acylated with acetyl, propionyl or butyryl groups, they markedly recovered both of their serological reactivities. These results are compatible with the interpretation that the Inaba antigen factor C possessed by the four bacteria is substantially related to the common presence of N-acyl groups, regardless of their identity, residing in the perosamine residues constituting the O polysaccharide chain of their LPS. It was also indicated that the group antigen factor A of O1 V. cholerae is substantially related to the 3-deoxy-L-glycerotetronyl groups residing in the perosamine homopolymer of Inaba LPS.
antigen, LPS, O-antigen, Vibrio, Vibrio cholerae O1
NCBI PubMed ID: 7538078Publication DOI: 10.1111/j.1432-1033.1995.0583k.xJournal NLM ID: 0107600Publisher: Oxford, UK: Blackwell Science Ltd. on behalf of the Federation of European Biochemical Societies
Institutions: Department of Microbiology, School of Pharmaceutical Sciences Josai University, Saitama, Japan, Division of Microbiology, National Institute of Hygienic Science, Tokyo, Japan
Methods: 13C NMR, 1H NMR, methylation, gel filtration, GC-MS, GLC, serological methods
- Article ID: 308
Lin M, Nielsen K "Binding of the Brucella abortus lipopolysaccharide O-chain fragment to a monoclonal antibody - Quantitative analysis by fluorescence quenching and polarization" -
Journal of Biological Chemistry 272(5) (1997) 2821-2827
An antigenic O-chain polysaccharide fragment derived from Brucella abortus lipopolysaccharide was labeled with 14.8 +/- 1.8 (n = 5) and 52.3 +/- 2.4 (n = 3) micromol of fluorescein/g of polysaccharide (designated FL1 and FL2, respectively) for use in investigating the binding of O-chain to a specific murine antibody YsT9 under equilibrium conditions. Upon binding to YsT9, the fluorescence of FL1 and FL2 was quenched 45-57% with no shift in the excitation and emission spectra, and polarization of fluorescence increased by 300-335%. With fluorescence quenching and polarization as sensitive signals for antibody-bound labeled O-chains, the equilibrium constants for binding of FL1, FL2, and unlabeled O-chain to YsT9 were determined to be within a similar order (1.5 x 10(7) to 2.0 x 10(7) M-1) using a nonlinear curve fitting approach rather than Scatchard analysis. These results indicated that covalent attachment of fluorescein groups to the O-chain did not influence the recognition of the YsT9-defined epitope by the antibody. The reversibility of the O-chain-antibody reaction was also demonstrated by showing a rapid depolarization of the labeled O-chain-antibody complex in the presence of unlabeled O-chain, suggesting that this displacement experiment could be exploited to quantify the Brucella polysaccharide antigen. The study described here provides a useful model for characterization of the complex formation between a carbohydrate-binding protein and a carbohydrate ligand and also for the design of a homogeneous assay system to quantitate antigens or antibodies of clinical interest.
Lipopolysaccharide, monoclonal antibodies, Brucella, Brucella abortus, binding, fluorescence, polarization, quantitative
NCBI PubMed ID: 9006923Journal NLM ID: 2985121RPublisher: Baltimore, MD: American Society for Biochemistry and Molecular Biology
Institutions: Animal Diseases Research Institute, Nepean, ON, Canada
- 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
- Article ID: 605
Aragón V, Díaz R, Moreno E, Moriyón I "Characterization of Brucella abortus and Brucella melitensis native haptens as outer membrane O-type polysaccharides independent from the smooth lipopolysaccharide" -
Journal of Bacteriology 178(4) (1996) 1070-1079
Brucella native haptens (NHs) extracted with hot water from smooth (S)-type B. abortus and B. melitensis were purified to high levels of serological activity and compared with the polysaccharide obtained by acid hydrolysis (PS) of the S lipopolysaccharide (S-LPS). By 13C nuclear magnetic resonance analysis, NHs showed the spectrum of a homopolymer of a-1,2- or a-1,2- plus a-1,3-linked 4-formamido-4,6-dideoxy-D-mannose (N-formylperosamine) previously reported for the LPS O chain. However, while PS contained up to 0.6% 3-deoxy-D-manno-2-octulosonate, this LPS-core marker was absent from NH. High performance liquid chromatography and thin-layer chromatography showed heterogeneity in NH purified from whole cells but not in PS. By immunoprecipitation, polysaccharides indistinguishable from NH were demonstrated in extracts obtained with phenol-water, saline at 60 C, and ether-water treatments, and none of these treatments caused S-LPS hydrolysis detectable with antibodies to the O chain and lipid A. Two lines of evidence showed that NH was in the cell surface. First, NH became biotinylated when B. abortus live cells were labelled with biotin-hydrazide, and the examination of cell fractions and electron microscopy sections with streptavidin-peroxidase and streptavidin-coloidal gold, respectively, showed that labelling was extrinsic. Moreover, whereas only traces of NH were found in cytosols, the amount of NH was enriched in cell envelopes and in the outer membrane blebs spontaneously released by brucellae during growth. Interactions between NH and S-LPS were observed in crude cell extracts, and such interactions could be reconstituted by using purified NH and LPS. The results demonstrate that NH is not a hydrolytic product of S-LPS and suggest a model in which LPS-independent O-type polysaccharides (NH) are intertwined with the O chain in the outer membrane of S-type brucellae.
Lipopolysaccharide, LPS, characterization, polysaccharide, polysaccharides, Brucella, Brucella abortus, hapten, Brucella melitensis, membrane, outer membrane, native, Haptens, O-type, smooth
NCBI PubMed ID: 8576040Journal NLM ID: 2985120RPublisher: American Society for Microbiology
Correspondence: iMORiYON@MAiL2.CTi.UNAV.ES
Institutions: Departamento de Microbiologia, Clinica Universitaria y Facultad de Medicina, Universidad de Navarra, Pamplona, Spain, Programa de Investigacion en Enfermedades Tropicales (PIEt), Escuela de Medicina Veterinaria, Universidad Nacional, Heredia, Costa Rica
Methods: gel filtration, NMR, SDS-PAGE, HPLC, immunoblotting, MAb studies, gel immunoprecipitation, HPTLC, extrinsic labelling
- Article ID: 774
Cloeckaert A, Weynants V, Godfroid J, Verger JM, Grayon M, Zygmunt MS "O-Polysaccharide epitope heterogeneity at the surface of Brucella spp. srudied by enzyme-linked immunosorbent assay and flow cytometry" -
Clinical and Diagnostic Laboratory Immunology 5(6) (1998) 862-870
Smooth Brucella strains are classified into three serotypes, i.e., A+M-, A-M+, and A+M+, according to slide agglutination with A and M monospecific polyclonal sera. The epitopes involved have been located on the O-polysaccharide (O-PS) moiety of the smooth lipopolysaccharide (S-LPS), which represents the most exposed antigenic structure on the surface of Brucella spp. By use of monoclonal antibodies (MAbs) a number of epitope specificities on the O-PS have been reported: A, M, and epitopes shared by both A and M dominant strains, which have been named common (C) epitopes. The latter have been further subdivided, according to relative MAb binding in enzyme-linked immunosorbent assays (ELISA) to A- and M-dominant Brucella strains and to cross-reacting Yersinia enterocolitica O:9, into five epitopic specificities: C (M>A), C (M=A), C/Y (M>A), C/Y (M=A), and C/Y (A>M). In the present study, we studied the occurrence of these epitopes at the surface of representatives of all Brucella species and biovars including the live vaccine strains by analyzing the levels of MAb binding to whole Brucella cells in ELISA and flow cytometry assays. In ELISA, the level of MAb binding correlated well with the previously defined epitope specificity and the serotype defined by polyclonal sera for each Brucella species, biovar, or strain. However, MAbs to the C (M=A) and C (M>A) epitopes showed insignificant binding to B. suis biovar 2 strains and bound at lower titers to B. suis biovar 3 and B. neotomae than to the other Brucella strains. Some of the flow cytometry results were contradictory to those obtained by ELISA. In fact, it appeared by flow cytometry that all O-PS epitopes, including the A and M epitopes, are shared to different degrees by Brucella spp. which nevertheless show a high degree of O-PS heterogeneity according to MAb binding intensities. The subdivision of MAb specificities and Brucella serotypes was therefore less evident by flow cytometry than by ELISA. Whereas in ELISA the MAb specific for the A epitope showed insignificant binding to Y. enterocolitica O:9, this MAb bound strongly to Y. enterocolitica O:9 in flow cytometry. One of the two MAbs specific to the C (M=A) epitope also bound at a low but significant level to B. suis biovar 2 strains. However, as in ELISA the MAb specific for the C (M>A) epitope did not bind at all to B. suis biovar 2 strains in flow cytometry. Flow cytometry provided new information regarding specificity of the MAbs and may further explain some aspects of the capacity of passive protection of some MAbs against smooth Brucella infection in mice. As shown in the present study the occurrence of Brucella strains apparently completely devoid of one specific C O-PS epitope (e.g., B. suis biovar 2 devoid of the C [M>A] epitope) offers the possibility of obtaining vaccine strains devoid of a diagnostic O-PS epitope, which could further help to resolve the problem of discriminating infected from vaccinated animals that remains a major goal in brucellosis research.
strain, characterization, epitope, O-polysaccharide, O polysaccharide, specific, surface, Brucella, Yersinia enterocolitica, assay, enzyme-linked immunosorbent assay, flow cytometry, heterogeneity, immunosorbent
NCBI PubMed ID: 9801349Journal NLM ID: 9421292Publisher: Washington, DC: American Society for Microbiology
Correspondence: Cloeckaert@tours.inra.fr
Institutions: Institut National de la Recherche Agronomique, Laboratoire de Pathologie Infectieuse et Immunologie, 37380 Nouzilly, France, Unite d'Immunologie-Microbiologie, Facultes Universitaires Notre-Dame de la Paix, Namur, Centre d'Etude et de Recherches Veterinaires et Agrochimiques, B-1180 Brussels,3 Belgium
Methods: serological methods
- Article ID: 953
Lubeck PS, Hoorfar J, Ahrens P, Skurnik M "Cloning and characterization of the Yersinia enterocolitica serotype O:9 lipopolysaaccharide O-antigen gene cluster" -
Advances in Experimental Medicine and Biology 529 (2003) 207-210
no abstract
Lipopolysaccharide, biosynthesis, antigen, lipopolysaccharides, structure, core, gene, strain, characterization, serotype, O-antigen, O antigen, cell, cloning, cluster, gene cluster, cell wall, PAGE, Yersinia pseudotuberculosis, function, genus, bacteriophage, Yersinia, Yersinia enterocolitica, Yersinia pestis, influence, Bacteriophages, growth, temperature
NCBI PubMed ID: 12756758Publication DOI: 10.1007/0-306-48416-1_40Journal NLM ID: 0121103Publisher: Kluwer Academic/Plenum Publishers
Institutions: Department of Bacteriology, Danish Veterinary Institute, Copenhagen, Denmark, Department of Medical Biochemistry, University of Turku, Turku, Finland.
- Article ID: 1537
Lapaque N, Moriyón I, Moreno E, Gorvel JP "Brucella lipopolysaccharide acts as a virulence factor" -
Current Opinion in Microbiology 8(1) (2005) 60-66
Brucella is a facultative intracellular bacterium responsible for brucellosis. Virulence factors involved in Brucella replication and Brucella's strategies to circumvent the immune response are under investigation. VirB proteins that form the type IV secretion system and that are involved in intracellular replication are considered as one of Brucella's virulence factors. In addition to this secretion system, bacterial outer membrane components have also been described as being implicated in Brucella survival in the host. For example, this bacterium possesses an unconventional non-endotoxic lipopolysaccharide that confers resistance to anti-microbial attacks and modulates the host immune response. These properties make lipopolysaccharide an important virulence factor for Brucella survival and replication in the host
Lipopolysaccharide, Bacterial, host, virulence, form, property, type, factor, protein, response, bacteria, Brucella, intracellular, component, resistance, case, membrane, proteins, immune response, outer membrane, virulence factor, immune, France, secretion, antimicrobial, brucellosis, survival
NCBI PubMed ID: 15694858Journal NLM ID: 9815056Publisher: London; New York: Current Biology
Institutions: Centre d'Immunologie INSERM-CNRS-Universite de la Mediterranee, Parc Scientifique de Luminy, Case 906, 13288 Marseille 9, France
- 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: 1828
L'vov VL, Malikov VE, Shashkov AS, Dranovskaya EA, Dmitriev BA "Brucella somatic antigens. The structure of the O-specific polysaccharide chain of Brucella melitensis lipopolysaccharide" -
Bioorganicheskaya Khimia = Bioorganic Chemistry [Russian] 11(7) (1985) 963-969
The phenol-phase soluble antigenic lipopolysaccharide was isolated from Brucella melitensis, strain 565, by the routine phenol/water procedure followed by chromatography on Sepharose 4B. After mild acid hydrolysis and chromatography on Sephadex G-50, the lipopolysaccharide yielded a linear O-specific polysaccharide built up from 1,2-linked 4,6-dideoxy-4-formamido-α-D-mannopyranosyl units. The structure of the polysaccharide was deduced mainly from the nuclear magnetic resonance and methylation analyses. The phenol-soluble lipopolysaccharide, isolated from commercial vaccine strain B. abortus 19-BA, on mild hydrolysis afforded material, 13C and 1H-NMR spectra of which were identical to those of the O-specific polysaccharide from B. melitensis 565.
NCBI PubMed ID: 2413867Journal NLM ID: 7804941Publisher: Moskva: Nauka
Institutions: N.D. Zelinsky Institute of Organic Chemistry, Academy of Sciences of the USSR, Moscow, Russia
Methods: 13C NMR, 1H NMR
- Article ID: 2378
Caroff M, Bundle DR, Perry MB "Structure of the O-chain of the phenol-phase soluble cellular lipopolysaccharide of Yersinia enterocolitica serotype O:9" -
European Journal of Biochemistry 139 (1984) 195-200
The phenol-phase soluble cellular lipopolysaccharide isolated by the phenol/water extraction method from Yersinia enterocolitica serotype O:9 cells was shown by hydrolytic, periodate oxidation, methylation and nuclear magnetic resonance studies to be an S-type lipopolysaccharide with a linear O-antigenic polysaccharide of 1,2-linked 4,6-dideoxy-4-formamido-α-D-mannopyranosyl units. The serological cross-reactivity between Y. enterocolitica serotype O:9 and the lipopolysaccharides of Vibrio cholerae and Brucella species can now be related to the presence of N-acylated 4-amino-4,6-dideoxy-α-D-mannopyranosyl residues in their respective O-antigenic chains.
NCBI PubMed ID: 6199199Publication DOI: 10.1111/j.1432-1033.1984.tb07994.xJournal NLM ID: 0107600Publisher: Oxford, UK: Blackwell Science Ltd. on behalf of the Federation of European Biochemical Societies
Institutions: Division of Biological Sciences, National Research Council of Canada, 100 Sussex Drive, Ottawa, Ontario, Canada, K1A OR6
Methods: 13C NMR, 1H NMR, methylation, periodate oxidation, sugar analysis, MS
- Article ID: 2533
Caroff M, Bundle DR, Perry MB, Cherwonogrodzky JW, Duncan JR "Antigenic S-type lipopolysaccharide of Brucella abortus 1119-3" -
Infection and Immunity 46 (1984) 384-388
Antigenic phenol-phase soluble lipopolysaccharide isolated from Brucella abortus 1119-3 by hot phenol-water extraction was shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, controlled hydrolysis, periodate oxidation, methylation, and 1H and 13C nuclear magnetic resonance studies to be an S-type lipopolysaccharide which could be cleaved to yield a lipid A and an O-chain polysaccharide identified as an unbranched linear homopolymer of 1,2-linked 4,6-dideoxy-4-formamido-α-D-mannopyranosyl residues. The serological reactivity of bovine antiserum to B. abortus 1119-3 with the lipopolysaccharides of Yersinia enterocolitica serotype O:9 and Vibrio cholerae species has now been related to the occurrence of 1,2-linked N-acylated 4-amino-4,6-dideoxy-α-D-mannopyranosyl units in the O-chain polysaccharides of their lipopolysaccharides.
NCBI PubMed ID: 6437981Journal NLM ID: 0246127Publisher: American Society for Microbiology
Methods: 13C NMR, 1H NMR, methylation, periodate oxidation, SDS-PAGE, sugar analysis
- Article ID: 2745
Altman E, Bundle DR "Polysaccharide affinity columns for purification of lipopolysaccharide-specific murine monoclonal antibodies" -
Methods in Enzymology 247 (1994) 243-253
NCBI PubMed ID: 7534862Journal NLM ID: 0212271Institutions: Institute for Biological Sciences, National Research Council of Canada, Ottawa, Ontario
- Article ID: 2804
Bundle DR "Antibody combining sites and oligosaccharide determinants studied by competitive binding, sequencing, and X-ray crystallography" -
Pure and Applied Chemistry 61 (1989) 1171-1180
Synthetic oligosaccharides and polysaccharides of known structure have been covalently attached to carrier protein and enzymes for use in a strategy to generate and select monoclonal antibodies with well defined carbohydrate binding profiles. Direct competitive binding assays utilizing antibody bound to a solid phase and saccharide - enzyme conjugates provided convenient and reliable measurement of specificity . Protocols were developed to prepare these carbohydrate - enzyme conjugates. Antibodies specific for the Brucella A and M polysaccharide/antigen were selected and characterized by these techniques. The crystal structure of an Fab fragment derived from one antibody has been solved at 2.7A resolution. Preliminary crystal structure data in conjunction with the amino acid sequence of light and Fd polypeptide chains implicate tyrosine, arginine, asparagine, glutamine and aspartic acid as the residues that contact antigen. A second a antibody - antigen system involving the Shigella flexneri Y antigen has been studied by a similar approach, although in this case two distinct antibodies have been subjected to detailed binding studies with oligosaccharide fragments of the polysaccharide repeating unit and specifically modified derivatives. These data support the contention that the principal polar interactions between an oligosaccharide and its antibody involve a small group of hydroxyl residues and that specifically deoxygenated oligosaccharide inhibitors can have association constants significantly higher than the natural oligosaccharide.
Journal NLM ID: 0376514WWW link: http://pac.iupac.org/publications/pac/pdf/1989/pdf/6107x1171.pdfPublisher: Oxford: Blackwell Scientific Publications
Institutions: Division of Biological Science, National Research Council of Canada, Ottawa, Ontario, K1A OR6, Canada
- Article ID: 4307
Greenfield LK, Whitfield C "Synthesis of lipopolysaccharide O-antigens by ABC transporter-dependent pathways" -
Carbohydrate Research 356 (2012) 12-24
The O-polysaccharide (O-PS; O-antigen) of bacterial lipopolysaccharides is made up of repeating units of one or more sugar residues and displays remarkable structural diversity. Despite the structural variations, there are only three strategies for O-PS assembly. The ATP-binding cassette (ABC)-transporter-dependent mechanism of O-PS biosynthesis is widespread. The Escherichia coli O9a and Klebsiella pneumoniae O2a antigens provide prototypes, which are distinguished by the fine details that link glycan polymerization and chain termination at the cytoplasmic face of the inner membrane to its export via the ABC transporter. Here, we describe the current understanding of these processes. Since glycoconjugate assembly complexes that utilize an ABC transporter-dependent pathway are widespread among the bacterial kingdom, the models described here are expected to extend beyond O-PS biosynthesis systems
Lipopolysaccharide, O-polysaccharide, ATP-binding cassette transporter, Escherichia coli O9a, Klebsiella pneumoniae O2a
NCBI PubMed ID: 22475157Publication DOI: 10.1016/j.carres.2012.02.027Journal NLM ID: 0043535Publisher: Elsevier
Correspondence: C. Whitfield
Institutions: Department of Molecular and Cellular Biology, University of Guelph, Guelph, Ontario, Canada N1G 2W1
- Article ID: 4329
Knirel YA "Structure of O-antigens" -
Book: Bacterial lipopolysaccharides: Structure, chemical synthesis, biogenesis and interaction with host cells (2011) Chapter 3, 41-115
The lipopolysaccharide (LPS) is the major constituent of the outer leaflet of the outer membrane of Gram-negative bacteria. Its lipid A moiety is embedded in the membrane and serves as an anchor for the rest of the LPS molecule. The outermost repetitive glycan region of the LPS is linked to the lipid A through a core oligosaccharide (OS), and is designated as the O-specific polysaccharide (O-polysaccharide, OPS) or O-antigen. The O-antigen is the most variable portion of the LPS and provides serological specificity, which is used for bacterial serotyping. The OPS also provides protection to the microorganisms from host defenses such as complement mediated killing and phagocytosis, and is involved in interactions of bacteria with plants and bacteriophages. Studies of the OPSs ranging from the elucidation of their chemical structures and conformations to their biological and physico-chemical properties help improving classification schemes of Gram-negative bacteria. Furthermore, these studies contributed to a better understanding of the mechanisms of pathogenesis of infectious diseases, as well as provided information to develop novel vaccines and diagnostic reagents.
Lipopolysaccharide, synthesis, lipopolysaccharides, structure, Bacterial, host, O-antigen, O antigen, cell, O antigens, O-antigens, chemical, interaction, cells, PDF, chemical synthesis, biogenesis
Publication DOI: 10.1007/978-3-7091-0733-1_3Publisher: Springer
Correspondence: knirel@ioc.ac.ru
Editors: Knirel YA, Valvano MA
Institutions: Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia
- Article ID: 4560
Kubler-Kielb J, Vinogradov E "Reinvestigation of the structure of Brucella O-antigens" -
Carbohydrate Research 378 (2013) 144-147
O-Specific polysaccharides of Brucella contain two antigenic determinants, called A and M. Most of the strains express epitope A with a small amount of epitope M, whereas Brucella melitensis strain 16M expresses longer polymer consisting mostly of M-type epitopes. Proposed explanation was that epitope A is defined by 1-2-linked homopolymer of N-formylperosamine (Rha4NFo), while epitope M is a pentasaccharide with four 2- and one 3-substituted Rha4NFo. We reinvestigated both types of structures by 2D NMR and showed that M-epitope is a tetrasaccharide, missing one of the 2-linked Rha4NFo as compared to the previously proposed structure. Polysaccharide from B. melitensis 16M contains a fragment of 1-2-linked polymer, capped with M-type polymer. Other strains contain one or two M-type units at the non-reducing end of the 1-2-linked O-chain.
NMR, LPS, structure, O-antigens, O-specific, O-specific polysaccharide, Brucella, MS, Brucella melitensis
NCBI PubMed ID: 23664729Publication DOI: 10.1016/j.carres.2013.03.021Journal NLM ID: 0043535Publisher: Elsevier
Correspondence: E. Vinogradov
Institutions: Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD, USA
Methods: 13C NMR, 1H NMR, NMR-2D, GC-MS, sugar analysis, MALDI-TOF MS, NMR-1D, N-acetylation, de-N-acelation
- Article ID: 4644
Zaccheus MV, Ali T, Cloeckaert A, Zygmunt MS, Weintraub A, Iriarte M, Moriyón I, Widmalm G "The Epitopic and Structural Characterization of Brucella suis Biovar 2 O-Polysaccharide Demonstrates the Existence of a New M-Negative C-Negative Smooth Brucella Serovar" -
PLoS One 8(1) (2013) e53941
The brucellae are Gram-negative bacteria that cause an important zoonosis. Studies with the main Brucella species have shown that the O-antigens of the Brucella smooth lipopolysaccharide are α-(1→2) and α-(1→3)-linked N-formyl-perosamine polysaccharides that carry M, A and C (A = M, A>M and AA) and M specificities. However, the biovar 2 O-antigen bound monoclonal antibodies to the Brucella A epitope, and to the C/Y epitope shared by brucellae and Yersinia enterocolitica O:9, a bacterium that carries an N-formyl-perosamine O-antigen in exclusively α-(1→2)-linkages. By (13)C NMR spectroscopy, B. suis biovar 1 but not B. suis biovar 2 or Y. enterocolitica O:9 polysaccharide showed the signal characteristic of α-(1→3)-linked N-formyl-perosamine, indicating that biovar 2 may altogether lack this linkage. Taken together, the NMR spectroscopy and monoclonal antibody analyses strongly suggest a role for α-(1→3)-linked N-formyl-perosamine in the C (A = M) and C (M>A) epitopes. Moreover, they indicate that B. suis biovar 2 O-antigen lacks some lipopolysaccharide epitopes previously thought to be present in all smooth brucellae, thus representing a new brucella serovar that is M-negative, C-negative. Serologically and structurally this new serovar is more similar to Y. enterocolitica O:9 than to other brucellae.
Lipopolysaccharide, O-antigen, epitope, monoclonal antibodies, typing, Brucella suis
NCBI PubMed ID: 23335981Publication DOI: 10.1371/journal.pone.0053941Journal NLM ID: 101285081Publisher: San Francisco, CA: Public Library of Science
Correspondence: IM
; GW
Institutions: Department of Organic Chemistry, Arrhenius Laboratory, Stockholm University, Stockholm, Sweden
Methods: 13C NMR, 1H NMR, NMR-2D, DNA sequencing, ELISA, NMR-1D
- Article ID: 4983
Casabuono AC, Czibener C, Del Giudice MG, Valguarnera E, Ugalde JE, Couto AS "New Features in the Lipid A Structure of Brucella suis and Brucella abortus Lipopolysaccharide" -
Journal of the American Society for Mass Spectrometry 28(12) (2017) 2716-2723
Brucellaceae are Gram-negative bacteria that cause brucellosis, one of the most distributed worldwide zoonosis, transmitted to humans by contact with either infected animals or their products. The lipopolysaccharide exposed on the cell surface has been intensively studied and is considered a major virulence factor of Brucella. In the last years, structural studies allowed the determination of new structures in the core oligosaccharide and the O-antigen of this lipopolysaccharide. In this work, we have reinvestigated the lipid A structure isolated from B. suis and B. abortus lipopolysaccharides. A detailed study by MALDI-TOF mass spectrometry in the positive and negative ion modes of the lipid A moieties purified from both species was performed. Interestingly, a new feature was detected: the presence of a pyrophosphorylethanolamine residue substituting the backbone. LID-MS/MS analysis of some of the detected ions allowed assurance that the Lipid A structure composed by the diGlcN3N disaccharide, mainly hexa-acylated and penta-acylated, bearing one phosphate and one pyrophosphorylethanolamine residue. Graphical abstract
Lipopolysaccharide, lipid A, Brucella, Brucella abortus, MALDI-TOF MS, brucellosis, Brucella suis
NCBI PubMed ID: 28924631Publication DOI: 10.1007/s13361-017-1805-xJournal NLM ID: 9010412Publisher: Elsevier
Correspondence: acouto@qo.fcen.uba.ar
Institutions: Instituto de Investigaciones Biotecnologicas 'Dr. Rodolfo A. Ugalde', IIB-INTECH, CONICET, Universidad Nacional de San Martin, San Martin, Buenos Aires, Argentina, Universidad de Buenos Aires, Facultad de Ciencias Exactas y Naturales, Departamento de Quimica Organica - Consejo Nacional de Investigaciones Cientificas y Tecnicas, Centro de Investigacion en Hidratos de Carbono (CIHIDECAR), Ciudad Universitaria, Intendente Guiraldes 2160, C1428GA, Buenos Aires, Argentina
Methods: SDS-PAGE, acid hydrolysis, UV-MALDI-TOF MS, MALDI-LID-MS/MS
- Article ID: 5062
Bundle DR, McGiven J "Brucellosis: Improved Diagnostics and Vaccine Insights from Synthetic Glycans" -
Accounts of Chemical Research 50(12) (2017) 2958-2967
Brucellosis is a serious zoonotic bacterial disease that is ranked by the World Health Organization among the top seven "neglected zoonoses" that threaten human health and cause poverty. It is a costly, highly contagious disease that affects ruminants, cattle, sheep, goats, and other productive animals such as pigs. Symptoms include abortions, infertility, decreased milk production, weight loss, and lameness. Brucellosis is also the most common bacterial disease that is transmitted from animals to humans, with approximately 500 000 new human cases each year. Detection and slaughter of infected animals is required to eradicate the disease, as vaccination alone is currently insufficient. However, as the most protective vaccines compromise serodiagnosis, this creates policy dilemmas, and these often result in the failure of eradication and control programs. Detection of antibodies to the Brucella bacterial cell wall O-polysaccharide (OPS) component of smooth lipopolysaccharide is used in diagnosis of this disease, and the same molecule contributes important protective efficacy to currently deployed veterinary whole-cell vaccines. This has set up a long-standing paradox that while Brucella OPS confers protective efficacy to vaccines, its presence results in similar antibody profiles in infected and vaccinated animals. Consequently, differentiation of infected from vaccinated animals (DIVA) is not possible, and this limits efforts to combat the disease. Recent clarification of the chemical structure of Brucella OPS as a block copolymer of two oligosaccharide sequences has provided an opportunity to utilize unique oligosaccharides only available via chemical synthesis in serodiagnostic tests for the disease. These oligosaccharides show excellent sensitivity and specificity compared with the native polymer used in current commercial tests and have the added advantage of assisting discrimination between brucellosis and infections caused by several bacteria with OPS that share some structural features with those of Brucella. During synthesis and immunochemical evaluation of these synthetic antigens, it became apparent that an opportunity existed to create a polysaccharide-protein conjugate vaccine that would not create antibodies that give false positive results in diagnostic tests for infection. This objective was reduced to practice, and immunization of mice showed that antibodies to the Brucella A antigen could be developed without reacting in a diagnostic test based on the M antigen. A conjugate vaccine of this type could readily be developed for use in humans and animals. However, as chemical methods advance and modern methods of bacterial engineering mature, it is expected that the principles elucidated by these studies could be applied to the development of an inexpensive and cost-effective vaccine to combat endemic brucellosis in animals.
O-polysaccharide, Brucella, vaccine
NCBI PubMed ID: 29219305Publication DOI: 10.1021/acs.accounts.7b00445Journal NLM ID: 0157313Publisher: Washington, DC: American Chemical Society
Correspondence: dave.bundle@ualberta.ca
Institutions: Department of Chemistry, University of Alberta, Edmonton, AB, Canada, FAO/WHO Collaborating Centre for Brucellosis, OIE Brucellosis Reference Laboratory, Department of Bacteriology, Animal & Plant Health Agency , Woodham Lane, Addlestone, Surrey KT15 3NB, United Kingdom
Methods: chemical synthesis, glycosylation, immunization, conjugation to tetanus toxoid
- Article ID: 5185
Martinez-Gomez E, Stahle J, Gil-Ramirez Y, Zuniga-Ripa A, Zaccheus M, Moriyón I, Iriarte M, Widmalm G, Conde-Alvarez R "Genomic Insertion of a Heterologous Acetyltransferase Generates a New Lipopolysaccharide Antigenic Structure in Brucella abortus and Brucella melitensis" -
Frontiers in Microbiology 9 (2018) 1092
Brucellosis is a bacterial zoonosis of worldwide distribution caused by bacteria of the genus Brucella. In Brucella abortus and Brucella melitensis, the major species infecting domestic ruminants, the smooth lipopolysaccharide (S-LPS) is a virulence factor. This S-LPS carries a N-formyl-perosamine homopolymer O-polysaccharide that is the major antigen in serodiagnostic tests and is required for virulence. We report that the Brucella O-PS can be structurally and antigenically modified using wbdR, the acetyl-transferase gene involved in N-acetyl-perosamine synthesis in Escherichia coli O157:H7. Brucella constructs carrying plasmidic wbdR expressed a modified O-polysaccharide but were unstable, a problem circumvented by inserting wbdR into a neutral site of chromosome II. As compared to wild-type bacteria, both kinds of wbdR constructs expressed shorter O-polysaccharides and NMR analyses showed that they contained both N-formyl and N-acetyl-perosamine. Moreover, deletion of the Brucella formyltransferase gene wbkC in wbdR constructs generated bacteria producing only N-acetyl-perosamine homopolymers, proving that wbdR can replace for wbkC. Absorption experiments with immune sera revealed that the wbdR constructs triggered antibodies to new immunogenic epitope(s) and the use of monoclonal antibodies proved that B. abortus and B. melitensis wbdR constructs respectively lacked the A or M epitopes, and the absence of the C epitope in both backgrounds. The wbdR constructs showed resistance to polycations similar to that of the wild-type strains but displayed increased sensitivity to normal serum similar to that of a per R mutant. In mice, the wbdR constructs produced chronic infections and triggered antibody responses that can be differentiated from those evoked by the wild-type strain in S-LPS ELISAs. These results open the possibilities of developing brucellosis vaccines that are both antigenically tagged and lack the diagnostic epitopes of virulent field strains, thereby solving the diagnostic interference created by current vaccines against Brucella.
antigen, Brucella, acetyltransferase, lipopolysaccharide (LPS), virulence factor, brucellosis, bacterial pathogenesis, vaccine development
NCBI PubMed ID: 29887851Publication DOI: 10.3389/fmicb.2018.01092Journal NLM ID: 101548977Publisher: Lausanne: Frontiers Research Foundation
Correspondence: Raquel Conde-Бlvarez
Institutions: Department of Organic Chemistry, Arrhenius Laboratory, Stockholm University, Stockholm, Sweden, Instituto de Salud Tropical, Instituto de Investigacion Sanitaria de Navarra, Departamento de Microbiologia y Parasitologia, Universidad de Navarra, Pamplona, Spain
Methods: 13C NMR, 1H NMR, NMR-2D, SDS-PAGE, DNA techniques, ELISA, mild acid hydrolysis, Western blotting, biological assays, serum bactericidal assays, immunological assays
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8. Compound ID: 867
|
-2)-a-D-Rhap4NFo-(1-2)-a-D-Rhap4NFo-(1-3)-a-D-Rhap4NFo-(1-2)-a-D-Rhap4NFo-(1-2)-a-D-Rhap4NFo-(1- |
Show graphically |
Structure type: polymer chemical repeating unit
Trivial name: M antigen
Compound class: O-polysaccharide, O-antigen
Contained glycoepitopes: IEDB_130647,IEDB_131172,IEDB_134279,IEDB_134280,IEDB_134281,IEDB_1397515,IEDB_2116320,IEDB_434544,IEDB_434545,IEDB_434546
The structure is contained in the following publication(s):
- Article ID: 241
Godfroid F, Cloeckaert A, Taminiau B, Danese I, Tibor A, De Bolle X, Mertens P, Letesson JJ "Genetic organisation of the lipopolysaccharide O-antigen biosynthesis region of Brucella melitensis 16M (wbk)" -
Research in Microbiology 151(8) (2000) 655-668
Brucella spp. are Gram-negative, facultative intracellular bacteria that cause a zoonotic world-wide disease. As in other Gram-negative bacteria, its S-LPS (smooth lipopolysaccharide) is a major determinant of virulence. The Brucella melitensis 16M LPS O-antigen is a homopolymer of 4-formamido-4,6, dideoxymannose. In this study, the previously cloned 14-kb wbk gene cluster was sequenced, and seven open reading frames (ORFs) as well as four insertion sequences were identified. Six of the seven ORFs are homologous to LPS biosynthesis genes from other organisms. The gmd, per and wbkC gene products are predicted to be involved in 4-formamido-4,6,dideoxymannose synthesis. By deletion experiments, we demonstrated that the putative formyltransferase WbkC is absolutely required for the O-side-chain production. The wbkA gene product is similar to several mannosyltransferases and is probably involved in the polymerisation of the B. melitensis O-side-chain. We also identified two genes (wzm and wzt) encoding proteins with high similarity to several two-component ABC (ATP-binding cassette) transporters. Their implication in O-antigen translocation across the inner membrane was confirmed by gene replacement. Finally, no function has been assigned to the wbkB gene either by homology search or functionally, because deletion of wbkB did not interfere with the O-antigen structure. The seven ORFs have a low G + C content, indicating that they might have been acquired by lateral transfer from a progenitor with more A + T rich DNA
Lipopolysaccharide, biosynthesis, genetic, O-antigen, O antigen, O-side-chain, region, Brucella, ABC transporter, Brucella melitensis, lipopolysaccharide O-antigen, LPS genetics
NCBI PubMed ID: 11081580Journal NLM ID: 8907468Publisher: Elsevier
Correspondence: Jean-jaques.letesson@fundp.ac.be
Institutions: Unite de recherche en biologie moleculaire (URBM), Laboratoire d'immunologie et de microbiologie, Facultes universitaires Notre Dame de la Paix, Belgium
- Article ID: 605
Aragón V, Díaz R, Moreno E, Moriyón I "Characterization of Brucella abortus and Brucella melitensis native haptens as outer membrane O-type polysaccharides independent from the smooth lipopolysaccharide" -
Journal of Bacteriology 178(4) (1996) 1070-1079
Brucella native haptens (NHs) extracted with hot water from smooth (S)-type B. abortus and B. melitensis were purified to high levels of serological activity and compared with the polysaccharide obtained by acid hydrolysis (PS) of the S lipopolysaccharide (S-LPS). By 13C nuclear magnetic resonance analysis, NHs showed the spectrum of a homopolymer of a-1,2- or a-1,2- plus a-1,3-linked 4-formamido-4,6-dideoxy-D-mannose (N-formylperosamine) previously reported for the LPS O chain. However, while PS contained up to 0.6% 3-deoxy-D-manno-2-octulosonate, this LPS-core marker was absent from NH. High performance liquid chromatography and thin-layer chromatography showed heterogeneity in NH purified from whole cells but not in PS. By immunoprecipitation, polysaccharides indistinguishable from NH were demonstrated in extracts obtained with phenol-water, saline at 60 C, and ether-water treatments, and none of these treatments caused S-LPS hydrolysis detectable with antibodies to the O chain and lipid A. Two lines of evidence showed that NH was in the cell surface. First, NH became biotinylated when B. abortus live cells were labelled with biotin-hydrazide, and the examination of cell fractions and electron microscopy sections with streptavidin-peroxidase and streptavidin-coloidal gold, respectively, showed that labelling was extrinsic. Moreover, whereas only traces of NH were found in cytosols, the amount of NH was enriched in cell envelopes and in the outer membrane blebs spontaneously released by brucellae during growth. Interactions between NH and S-LPS were observed in crude cell extracts, and such interactions could be reconstituted by using purified NH and LPS. The results demonstrate that NH is not a hydrolytic product of S-LPS and suggest a model in which LPS-independent O-type polysaccharides (NH) are intertwined with the O chain in the outer membrane of S-type brucellae.
Lipopolysaccharide, LPS, characterization, polysaccharide, polysaccharides, Brucella, Brucella abortus, hapten, Brucella melitensis, membrane, outer membrane, native, Haptens, O-type, smooth
NCBI PubMed ID: 8576040Journal NLM ID: 2985120RPublisher: American Society for Microbiology
Correspondence: iMORiYON@MAiL2.CTi.UNAV.ES
Institutions: Departamento de Microbiologia, Clinica Universitaria y Facultad de Medicina, Universidad de Navarra, Pamplona, Spain, Programa de Investigacion en Enfermedades Tropicales (PIEt), Escuela de Medicina Veterinaria, Universidad Nacional, Heredia, Costa Rica
Methods: gel filtration, NMR, SDS-PAGE, HPLC, immunoblotting, MAb studies, gel immunoprecipitation, HPTLC, extrinsic labelling
- Article ID: 774
Cloeckaert A, Weynants V, Godfroid J, Verger JM, Grayon M, Zygmunt MS "O-Polysaccharide epitope heterogeneity at the surface of Brucella spp. srudied by enzyme-linked immunosorbent assay and flow cytometry" -
Clinical and Diagnostic Laboratory Immunology 5(6) (1998) 862-870
Smooth Brucella strains are classified into three serotypes, i.e., A+M-, A-M+, and A+M+, according to slide agglutination with A and M monospecific polyclonal sera. The epitopes involved have been located on the O-polysaccharide (O-PS) moiety of the smooth lipopolysaccharide (S-LPS), which represents the most exposed antigenic structure on the surface of Brucella spp. By use of monoclonal antibodies (MAbs) a number of epitope specificities on the O-PS have been reported: A, M, and epitopes shared by both A and M dominant strains, which have been named common (C) epitopes. The latter have been further subdivided, according to relative MAb binding in enzyme-linked immunosorbent assays (ELISA) to A- and M-dominant Brucella strains and to cross-reacting Yersinia enterocolitica O:9, into five epitopic specificities: C (M>A), C (M=A), C/Y (M>A), C/Y (M=A), and C/Y (A>M). In the present study, we studied the occurrence of these epitopes at the surface of representatives of all Brucella species and biovars including the live vaccine strains by analyzing the levels of MAb binding to whole Brucella cells in ELISA and flow cytometry assays. In ELISA, the level of MAb binding correlated well with the previously defined epitope specificity and the serotype defined by polyclonal sera for each Brucella species, biovar, or strain. However, MAbs to the C (M=A) and C (M>A) epitopes showed insignificant binding to B. suis biovar 2 strains and bound at lower titers to B. suis biovar 3 and B. neotomae than to the other Brucella strains. Some of the flow cytometry results were contradictory to those obtained by ELISA. In fact, it appeared by flow cytometry that all O-PS epitopes, including the A and M epitopes, are shared to different degrees by Brucella spp. which nevertheless show a high degree of O-PS heterogeneity according to MAb binding intensities. The subdivision of MAb specificities and Brucella serotypes was therefore less evident by flow cytometry than by ELISA. Whereas in ELISA the MAb specific for the A epitope showed insignificant binding to Y. enterocolitica O:9, this MAb bound strongly to Y. enterocolitica O:9 in flow cytometry. One of the two MAbs specific to the C (M=A) epitope also bound at a low but significant level to B. suis biovar 2 strains. However, as in ELISA the MAb specific for the C (M>A) epitope did not bind at all to B. suis biovar 2 strains in flow cytometry. Flow cytometry provided new information regarding specificity of the MAbs and may further explain some aspects of the capacity of passive protection of some MAbs against smooth Brucella infection in mice. As shown in the present study the occurrence of Brucella strains apparently completely devoid of one specific C O-PS epitope (e.g., B. suis biovar 2 devoid of the C [M>A] epitope) offers the possibility of obtaining vaccine strains devoid of a diagnostic O-PS epitope, which could further help to resolve the problem of discriminating infected from vaccinated animals that remains a major goal in brucellosis research.
strain, characterization, epitope, O-polysaccharide, O polysaccharide, specific, surface, Brucella, Yersinia enterocolitica, assay, enzyme-linked immunosorbent assay, flow cytometry, heterogeneity, immunosorbent
NCBI PubMed ID: 9801349Journal NLM ID: 9421292Publisher: Washington, DC: American Society for Microbiology
Correspondence: Cloeckaert@tours.inra.fr
Institutions: Institut National de la Recherche Agronomique, Laboratoire de Pathologie Infectieuse et Immunologie, 37380 Nouzilly, France, Unite d'Immunologie-Microbiologie, Facultes Universitaires Notre-Dame de la Paix, Namur, Centre d'Etude et de Recherches Veterinaires et Agrochimiques, B-1180 Brussels,3 Belgium
Methods: serological methods
- Article ID: 1754
Bundle DR, Cherwonogrodzky JW, Perry MB "Structural elucidation of the Brucella melitensis M antigen by high resolution NMR at 500-MHz" -
Biochemistry 26 (1987) 8717-8726
The Brucella M antigen from the species type strain Brucella melitensis 16M has been identified as a component of the cell wall lipopolysaccharide (LPS). O polysaccharide liberated from this LPS by mild acid hydrolysis exhibited M activity in serological tests and was shown to be a homopolymer of 4-formamido-4,6-dideoxy-α-D-mannopyranosyl residues arranged in an oligosaccharide repeating unit as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the native lipopolysaccharide. Structural analysis of the O polysaccharide by NMR methods was difficult due to apparent microheterogeneity of the repeating unit, which was in fact caused by the presence of rotational isomers of the N-formyl moiety. This problem was resolved by chemical modification of the polysaccharide to its amino and N-acetyl derivatives, the 500-MHz 1H and 125-MHz 13C NMR spectra of which could be analyzed in terms of a unique structure through application of pH-dependent beta-shifts and two-dimensional techniques that included COSY, relayed COSY, and NOESY experiments together with heteronuclear C/H shift correlation spectroscopy. On the basis of these experiments and supported by methylation and periodate oxidation data, the structure of the M polysaccharide was determined as a linear polymer of unbranched pentasaccharide repeating units consisting of four 1,2-linked and one 1,3-linked 4,6-dideoxy-4-formamido-α-D-mannopyranosyl residues. The marked structural similarity of the M antigen and the A antigen, which is known to be a 1,2-linked homopolysaccharide of 4,6-dideoxy-4-formamido-α-D-mannopyranosyl units, accounts for cross-serological reactions of the two and the long-standing confusion surrounding the nature of their antigenic determinants. Structural and serological considerations in conjuction with the sodium dodecyl sulfate banding pattern of Brucella A LPS suggest that its biosynthesis differs appreciably from that of the M antigen, which appears to be synthesized by regulated assembly of preformed oligosaccharide repeating units. Temperature, lysogenic phage may be responsible for such biosynthetic and structural variations.
NCBI PubMed ID: 3442684Journal NLM ID: 0370623Publisher: American Chemical Society
Institutions: Division of Biological Science, National Research Council of Canada, Ottawa
Methods: 13C NMR, 1H NMR
- 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: 2535
Moreno E, Mayer H, Moriyón I "Characterization of a native polysaccharide hapten from Brucella melitensis" -
Infection and Immunity 55 (1987) 2850-2853
Journal NLM ID: 0246127Publisher: American Society for Microbiology
Methods: 13C NMR
- Article ID: 2706
Cloeckaert A, Zygmunt MS, Dubray G, Limet JN "Characterization of O-polysaccharide specific monoclonal antibodies derived from mice infected with the rough Brucella melitensis strain B115" -
Journal of General Microbiology 139 (1993) 1551-1556
- Article ID: 2804
Bundle DR "Antibody combining sites and oligosaccharide determinants studied by competitive binding, sequencing, and X-ray crystallography" -
Pure and Applied Chemistry 61 (1989) 1171-1180
Synthetic oligosaccharides and polysaccharides of known structure have been covalently attached to carrier protein and enzymes for use in a strategy to generate and select monoclonal antibodies with well defined carbohydrate binding profiles. Direct competitive binding assays utilizing antibody bound to a solid phase and saccharide - enzyme conjugates provided convenient and reliable measurement of specificity . Protocols were developed to prepare these carbohydrate - enzyme conjugates. Antibodies specific for the Brucella A and M polysaccharide/antigen were selected and characterized by these techniques. The crystal structure of an Fab fragment derived from one antibody has been solved at 2.7A resolution. Preliminary crystal structure data in conjunction with the amino acid sequence of light and Fd polypeptide chains implicate tyrosine, arginine, asparagine, glutamine and aspartic acid as the residues that contact antigen. A second a antibody - antigen system involving the Shigella flexneri Y antigen has been studied by a similar approach, although in this case two distinct antibodies have been subjected to detailed binding studies with oligosaccharide fragments of the polysaccharide repeating unit and specifically modified derivatives. These data support the contention that the principal polar interactions between an oligosaccharide and its antibody involve a small group of hydroxyl residues and that specifically deoxygenated oligosaccharide inhibitors can have association constants significantly higher than the natural oligosaccharide.
Journal NLM ID: 0376514WWW link: http://pac.iupac.org/publications/pac/pdf/1989/pdf/6107x1171.pdfPublisher: Oxford: Blackwell Scientific Publications
Institutions: Division of Biological Science, National Research Council of Canada, Ottawa, Ontario, K1A OR6, Canada
- Article ID: 4307
Greenfield LK, Whitfield C "Synthesis of lipopolysaccharide O-antigens by ABC transporter-dependent pathways" -
Carbohydrate Research 356 (2012) 12-24
The O-polysaccharide (O-PS; O-antigen) of bacterial lipopolysaccharides is made up of repeating units of one or more sugar residues and displays remarkable structural diversity. Despite the structural variations, there are only three strategies for O-PS assembly. The ATP-binding cassette (ABC)-transporter-dependent mechanism of O-PS biosynthesis is widespread. The Escherichia coli O9a and Klebsiella pneumoniae O2a antigens provide prototypes, which are distinguished by the fine details that link glycan polymerization and chain termination at the cytoplasmic face of the inner membrane to its export via the ABC transporter. Here, we describe the current understanding of these processes. Since glycoconjugate assembly complexes that utilize an ABC transporter-dependent pathway are widespread among the bacterial kingdom, the models described here are expected to extend beyond O-PS biosynthesis systems
Lipopolysaccharide, O-polysaccharide, ATP-binding cassette transporter, Escherichia coli O9a, Klebsiella pneumoniae O2a
NCBI PubMed ID: 22475157Publication DOI: 10.1016/j.carres.2012.02.027Journal NLM ID: 0043535Publisher: Elsevier
Correspondence: C. Whitfield
Institutions: Department of Molecular and Cellular Biology, University of Guelph, Guelph, Ontario, Canada N1G 2W1
- Article ID: 4500
Ciesielski F, Griffin DC, Rittig M, Moriyón I, Bonev BB "Interactions of lipopolysaccharide with lipid membranes, raft models - a solid state NMR study" -
Biochimica et Biophysica Acta 1828(8) (2013) 1731-1742
Lipopolysaccharide (LPS) is a major component of the external leaflet of bacterial outer membranes, key pro-inflammatory factor and an important mediator of host-pathogen interactions. In host cells it activates the complement along with a pro-inflammatory response via a TLR4-mediated signalling cascade and shows preference for cholesterol-containing membranes. Here, we use solid state (13)C and (31)P MAS NMR to investigate the interactions of LPS from three bacterial species, Brucella melitensis, Klebsiella pneumoniae and Escherichia coli, with mixed lipid membranes, raft models. All endotoxin types are found to be pyrophosphorylated and Klebsiellar LPS is phosphonylated, as well. Carbon-13 MAS NMR indicates an increase in lipid order in the presence of LPS. Longitudinal (31)P relaxation, providing a direct probe of LPS molecular and segmental mobility, reveals a significant reduction in (31)P T1 times and lower molecular mobility in the presence of ternary lipid mixtures. Along with the ordering effect on membrane lipid, this suggests a preferential partitioning of LPS into ordered bilayer sphingomyelin/cholesterol-rich domains. We hypothesise that this is an important evolutionary drive for the selection of GPI-anchored raft-associated LPS-binding proteins as a first line of response to membrane-associated LPS.
LPS, endotoxin, host-pathogen interactions, lipid domains, high resolution solid state NMR, longitudinal relaxation
NCBI PubMed ID: 23567915Publication DOI: 10.1016/j.bbamem.2013.03.029Journal NLM ID: 0217513Publisher: Elsevier
Correspondence: B.B. Bonev
Institutions: School of Biomedical Sciences, University of Nottingham, Nottingham NG7 2UH, UK, Departamento de Microbiología, Universidad de Navarra, e Instituto de Salud Tropical, 31008 Pamplona, Spain
Methods: NMR
- Article ID: 4560
Kubler-Kielb J, Vinogradov E "Reinvestigation of the structure of Brucella O-antigens" -
Carbohydrate Research 378 (2013) 144-147
O-Specific polysaccharides of Brucella contain two antigenic determinants, called A and M. Most of the strains express epitope A with a small amount of epitope M, whereas Brucella melitensis strain 16M expresses longer polymer consisting mostly of M-type epitopes. Proposed explanation was that epitope A is defined by 1-2-linked homopolymer of N-formylperosamine (Rha4NFo), while epitope M is a pentasaccharide with four 2- and one 3-substituted Rha4NFo. We reinvestigated both types of structures by 2D NMR and showed that M-epitope is a tetrasaccharide, missing one of the 2-linked Rha4NFo as compared to the previously proposed structure. Polysaccharide from B. melitensis 16M contains a fragment of 1-2-linked polymer, capped with M-type polymer. Other strains contain one or two M-type units at the non-reducing end of the 1-2-linked O-chain.
NMR, LPS, structure, O-antigens, O-specific, O-specific polysaccharide, Brucella, MS, Brucella melitensis
NCBI PubMed ID: 23664729Publication DOI: 10.1016/j.carres.2013.03.021Journal NLM ID: 0043535Publisher: Elsevier
Correspondence: E. Vinogradov
Institutions: Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD, USA
Methods: 13C NMR, 1H NMR, NMR-2D, GC-MS, sugar analysis, MALDI-TOF MS, NMR-1D, N-acetylation, de-N-acelation
- Article ID: 4983
Casabuono AC, Czibener C, Del Giudice MG, Valguarnera E, Ugalde JE, Couto AS "New Features in the Lipid A Structure of Brucella suis and Brucella abortus Lipopolysaccharide" -
Journal of the American Society for Mass Spectrometry 28(12) (2017) 2716-2723
Brucellaceae are Gram-negative bacteria that cause brucellosis, one of the most distributed worldwide zoonosis, transmitted to humans by contact with either infected animals or their products. The lipopolysaccharide exposed on the cell surface has been intensively studied and is considered a major virulence factor of Brucella. In the last years, structural studies allowed the determination of new structures in the core oligosaccharide and the O-antigen of this lipopolysaccharide. In this work, we have reinvestigated the lipid A structure isolated from B. suis and B. abortus lipopolysaccharides. A detailed study by MALDI-TOF mass spectrometry in the positive and negative ion modes of the lipid A moieties purified from both species was performed. Interestingly, a new feature was detected: the presence of a pyrophosphorylethanolamine residue substituting the backbone. LID-MS/MS analysis of some of the detected ions allowed assurance that the Lipid A structure composed by the diGlcN3N disaccharide, mainly hexa-acylated and penta-acylated, bearing one phosphate and one pyrophosphorylethanolamine residue. Graphical abstract
Lipopolysaccharide, lipid A, Brucella, Brucella abortus, MALDI-TOF MS, brucellosis, Brucella suis
NCBI PubMed ID: 28924631Publication DOI: 10.1007/s13361-017-1805-xJournal NLM ID: 9010412Publisher: Elsevier
Correspondence: acouto@qo.fcen.uba.ar
Institutions: Instituto de Investigaciones Biotecnologicas 'Dr. Rodolfo A. Ugalde', IIB-INTECH, CONICET, Universidad Nacional de San Martin, San Martin, Buenos Aires, Argentina, Universidad de Buenos Aires, Facultad de Ciencias Exactas y Naturales, Departamento de Quimica Organica - Consejo Nacional de Investigaciones Cientificas y Tecnicas, Centro de Investigacion en Hidratos de Carbono (CIHIDECAR), Ciudad Universitaria, Intendente Guiraldes 2160, C1428GA, Buenos Aires, Argentina
Methods: SDS-PAGE, acid hydrolysis, UV-MALDI-TOF MS, MALDI-LID-MS/MS
Expand this compound
Collapse this compound
9. Compound ID: 868
Structure type: homopolymer
Compound class: O-polysaccharide, O-antigen
The structure is contained in the following publication(s):
- Article ID: 242
Gonzalez L, Asensio JL, Ariosa-Alvarez A, Verez-Bencomo V, Jiménez-Barbero J "Solution conformation and dynamics of the trisaccharide fragments of the O-antigen of Vibrio cholerae O1, serotypes Inaba and Ogawa" -
Carbohydrate Research 321(1-2) (1999) 88-95
The conformational behavior of the trisaccharide fragments of the Ogawa and Inaba Vibrio cholerae serotypes has been studied using NMR and molecular dynamics (MD). The obtained results indicate that there are no significant differences in the major conformation and in the extent of motion of the glycosidic torsions of these molecules. The differences in biological activity are probably not due to conformational effects but to van der Waals and/or hydrogen bonding interactions between the antigens and the biological receptor.
NMR spectroscopy, Vibrio cholerae, Serotypes, Molecular mechanics
NCBI PubMed ID: 10612004Journal NLM ID: 0043535Publisher: Elsevier
Institutions: Departamento Quimica Organica Biologica, Instituto Quimica Organica, CSIC, Madrid, Spain
Methods: NMR
- Article ID: 269
Isshiki Y, Haishima Y, Kondo S, Hisatsune K "Immunochemistry of group A and Inaba C antigen factors constituting the O antigen of O1 Vibrio cholerae" -
European Journal of Biochemistry 229 (1995) 583-588
Serological cross-reactivity among intact lipopolysaccharides (LPS) from O1 Vibrio cholerae Inaba O-form (Inaba), Yersinia enterocolitica O9 (O9), non-O1 V. cholerae serogroup Hakata (Hakata) and Vibrio bio-serogroup 1875 Variant (1875 Variant) (all of which share Inaba antigen factor C), as well as a total of six kinds of chemically modified LPS (three from O9 and three from Inaba) was demonstrated by passive hemolysis and passive hemolysis inhibition by using these LPS as antigen for sensitizing sheep red blood cells and as inhibitor. These intact as well as chemically modified LPS contained, in their O polysaccharide chain, α(1→2)-linked linear perosamine (4-amino-4,6-dideoxy-D-manno-pyranose) homopolymers with different N-acyl groups: their acyl groups comprise 3-deoxy-L-glycero-tetronyl (Inaba LPS), formyl (O9 LPS), 3-hydroxypropionyl (1875 Variant LPS), acetyl (Hakata LPS and artificially introduced into Inaba and O9 LPS), propionyl and butyryl (both artificially introduced into Inaba and O9 LPS) groups. N-Deacylation of the α(l→2)-linked N-(3-deoxy-L-glycero-tetronyl)perosamine homopolymer of Inaba and the N-formyl one of O9 LPS resulted in virtual elimination of their serological reactivity with both homologous and heterologous antisera. Furthermore, when the resultant NH2 groups of the N-deacylated perosamine homopolymers of both LPS were N-acylated with acetyl, propionyl or butyryl groups, they markedly recovered both of their serological reactivities. These results are compatible with the interpretation that the Inaba antigen factor C possessed by the four bacteria is substantially related to the common presence of N-acyl groups, regardless of their identity, residing in the perosamine residues constituting the O polysaccharide chain of their LPS. It was also indicated that the group antigen factor A of O1 V. cholerae is substantially related to the 3-deoxy-L-glycerotetronyl groups residing in the perosamine homopolymer of Inaba LPS.
antigen, LPS, O-antigen, Vibrio, Vibrio cholerae O1
NCBI PubMed ID: 7538078Publication DOI: 10.1111/j.1432-1033.1995.0583k.xJournal NLM ID: 0107600Publisher: Oxford, UK: Blackwell Science Ltd. on behalf of the Federation of European Biochemical Societies
Institutions: Department of Microbiology, School of Pharmaceutical Sciences Josai University, Saitama, Japan, Division of Microbiology, National Institute of Hygienic Science, Tokyo, Japan
Methods: 13C NMR, 1H NMR, methylation, gel filtration, GC-MS, GLC, serological methods
- Article ID: 394
Szu SC, Gupta R, Kovác P, Taylor DN, Robbins JB "Development of O-specific polysaccharide-protein conjugates is based upon the protective effect of serum vibriocidal antibodies against cholera" -
Bulletin de l'Institut Pasteur 93 (1995) 269-272
antibodies, O-specific, serum, conjugates, polysaccharide-protein conjugate, cholera, protective, vibriocidal
NCBI PubMed ID: 7582370Journal NLM ID: 0413647Publisher: Elsevier
Institutions: National Institutes of Health, Bethesda, MD 20892, Walter Reed Army Institute for Research, Washington, DC, 20014 (USA)
Methods: serological methods
- Article ID: 1046
Nesper J, Kapfhammer D, Klose KE, Merkert H, Reidl J "Characterization of Vibrio cholerae O1 antigen as the bacteriophage K139 receptor and identification of IS1004 insertions aborting O1 antigen biosynthesis" -
Journal of Bacteriology 182(18) (2000) 5097-5104
Bacteriophage K139 was recently characterized as a temperate phage of O1 Vibrio cholerae. In this study we have determined the phage adsorption site on the bacterial cell surface. Phage-binding studies with purified lipopolysaccharide (LPS) of different O1 serotypes and biotypes revealed that the O1 antigen serves as the phage receptor. In addition, phage-resistant O1 El Tor strains were screened by using a virulent isolate of phage K139. Analysis of the LPS of such spontaneous phage-resistant mutants revealed that most of them synthesize incomplete LPS molecules, composed of either defective O1 antigen or core oligosaccharide. By applying phage-binding studies, it was possible to distinguish between receptor mutants and mutations which probably caused abortion of later steps of phage infection. Furthermore, we investigated the genetic nature of O1-negative strains by Southern hybridization with probes specific for the O antigen biosynthesis cluster (rfb region). Two of the investigated O1 antigen-negative mutants revealed insertions of element IS1004 into the rfb gene cluster. Treating one wbeW::IS1004 serum-sensitive mutant with normal human serum, we found that several survivors showed precise excision of IS1004, restoring O antigen biosynthesis and serum resistance. Investigation of clinical isolates by screening for phage resistance and performing LPS analysis of nonlysogenic strains led to the identification of a strain with decreased O1 antigen presentation. This strain had a significant reduction in its ability to colonize the mouse small intestine.
biosynthesis, antigen, characterization, identification, insertion, Vibrio, Vibrio cholerae, Vibrio cholerae O1, bacteriophage, receptor
NCBI PubMed ID: 10960093Journal NLM ID: 2985120RPublisher: American Society for Microbiology
Correspondence: joachim.reidl@mail.uni-wuerzburg.de
Institutions: Zentrum fur Infektionsforschung1 and Institut fur Molekulare Infektionsforschung,3 Universitat Wurzburg, Wurzburg, Germany, Health Science Center, University of Texas, San Antonio, Texas 78284-77582
- Article ID: 1329
Zhang JA, Kovác P "Studies on vaccines against cholera. Synthesis of neoglycoconjugates from the hexasaccharide determinant of Vibrio cholerae O:1, serotype Ogawa, by single-point attachment or by attachment of the hapten in the form of clusters" -
Carbohydrate Research 321(3-4) (1999) 157-167
The terminal hexasaccharide of the O-antigen of Vibrio cholerae O:1, serotype Ogawa, has been synthesized in the form of a glycoside whose aglycon (linker) allows conjugation to carrier proteins by reductive amination. The conjugate obtained from direct, single-point attachment of the linker-equipped hapten to chicken serum albumin (CSA) contained seven hapten residues/CSA. A neoglycoconjugate containing the carbohydrate antigen in the form of clusters was obtained using, as a hapten subcarrier, an oligopeptide containing 16 amino groups. It was treated with a limited amount of hapten, to give a hapten-carrying subcarrier (HCS). Subsequent conjugation of HCS to CSA, using squaric acid diethyl ester as a conjugation reagent, gave a cross-linked, glycocluster conjugate containing 51% (w/w) of the carbohydrate
synthesis, oligosaccharide, hexasaccharide, serotype, form, cluster, vaccines, Vibrio, hapten, determinant, Vibrio cholerae, vaccine, cholera, glycoconjugate, attachment, neoglycoconjugate, glycocluster, subcarrier, synthetic vaccine
NCBI PubMed ID: 10614066Journal NLM ID: 0043535Publisher: Elsevier
Institutions: National Institutes of Health, NIDDK, Laboratory of Medicinal Chemistry, Bethesda, MD 20892-0815, USA
- Article ID: 1396
Bystricky S, Szu SC, Gotoh M, Kovác P "Circular dichroism of the O-specific polysaccharide of Vibrio cholerae O1 and some related derivatives" -
Carbohydrate Research 270(2) (1995) 115-122
The O-specific polysaccharide (O-SP) of Vibrio cholerae O1 is a homopolymer of α-(1→2)-linked 4-amino-4, 6-dideoxy-D-mannopyranose whose amino group is acylated with 3-deoxy-L-glycero-tetronic acid [N-(3-deoxy-L-glycero- tetronyl)-α-D-perosamine]. The circular dichroism (CD) of the O-SP as well as of a number of N-acyl (formyl, acetyl, 4-hydroxybutyl, 3-deoxy-L-and D-glycero-tetronyl) derivatives of methyl α-glycosides of 4-amino-4,6-dideoxy-D-mannopyranose (methyl α-D-perosaminide) has been studied for solutions in water, acetonitrile and 1,1,1-trifluoroethanol. The strong solvent dependence of the sign and intensity of the CD observed for the monosaccharide amides bearing achiral acyl groups is explained by solvent-mediated change of the orientation of the amido group relative to the proximal hydroxyl group at C-3. A change in the population of the nonplanar conformers with a pyramidal arrangement of bonds at the amido nitrogen has also been considered. The effect of solvents upon the CD spectra of compounds bearing chiral N-acyl substituents is less pronounced than that of their counterparts bearing achiral N-acyl substituents. The sign of the CD for the O-SP was found negative in all solvents used. This result is in agreement with the negative sign of the CD of the n → pi electron transition observed, independent of the solvent, for the monosaccharide derivative containing the L-glycero-3-deoxytetronamido group, and the positive sign found for its D-glycero-counterpart.
polysaccharide, O-specific, O-specific polysaccharide, derivative, Vibrio, Vibrio cholerae, Vibrio cholerae O1, circular dichroism, Vibrio cholerae O1NT:
NCBI PubMed ID: 7585695Journal NLM ID: 0043535Publisher: Elsevier
Institutions: NICHD, Laboratory of Developmental and Molecular Immunity, National Institutes of Health, Bethesda, USA, NICHD, Laboratory of Developmental and Molekular Immunity, National Institutes of Health, Bethesda, USA
Methods: CD
- Article ID: 1704
Hisatsune K, Kondo S, Isshiki Y, Iguchi T, Haishima Y "Occurrence of 2-O-methyl-N-(3-deoxy-L-glycero-tetronyl)-D-perosamine(4-amino-4,6-dideoxy-D-manno-pyranose) in lipopolysaccharide from Ogawa but not from Inaba O forms of O1 Vibrio cholerae" -
Biochemical and Biophysical Research Communications 190 (1993) 302-307
A structural study by GC-MS, methylation analysis, and 1H and 13C NMR was carried out on α (1→2)-linked linear N-(3-deoxy-L-glycero-tetronyl)-D-perosamine homopolymer constituting the O-polysaccharide chain of lipopolysaccharide from O1 Vibrio cholerae Ogawa and Inaba O forms. Occurrence of 2-O-methyl-N-(3-deoxy-L-glycero-tetronyl)-D-perosamine was demonstrated at the non-reducing terminus of the perosamine-homopolymer of lipopolysaccharide from the Ogawa O form in contrast to the presence of N-(3-deoxy-L-glycero-tetronyl)-D-perosamine at the nonreducing terminus for the Inaba O form
NCBI PubMed ID: 8422256Journal NLM ID: 0372516Publisher: Academic Press
Institutions: Department of Microbiology, School of Pharmaceutical Sciences, Josai University, Saitama, Japan
- 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: 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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10. Compound ID: 869
|
S-2,4HOBut-(1-4)-+
|
S-2,4HOBut-(1-4)-+ |
| |
S-2,4HOBut-(1-4)-a-D-Rhap4N-(1-2)-a-D-Rhap4N-(1-2)-a-D-Rhap4N-(1-1)-Me |
Show graphically |
Structure type: oligomer
The structure is contained in the following publication(s):
- Article ID: 242
Gonzalez L, Asensio JL, Ariosa-Alvarez A, Verez-Bencomo V, Jiménez-Barbero J "Solution conformation and dynamics of the trisaccharide fragments of the O-antigen of Vibrio cholerae O1, serotypes Inaba and Ogawa" -
Carbohydrate Research 321(1-2) (1999) 88-95
The conformational behavior of the trisaccharide fragments of the Ogawa and Inaba Vibrio cholerae serotypes has been studied using NMR and molecular dynamics (MD). The obtained results indicate that there are no significant differences in the major conformation and in the extent of motion of the glycosidic torsions of these molecules. The differences in biological activity are probably not due to conformational effects but to van der Waals and/or hydrogen bonding interactions between the antigens and the biological receptor.
NMR spectroscopy, Vibrio cholerae, Serotypes, Molecular mechanics
NCBI PubMed ID: 10612004Journal NLM ID: 0043535Publisher: Elsevier
Institutions: Departamento Quimica Organica Biologica, Instituto Quimica Organica, CSIC, Madrid, Spain
Methods: NMR
- Article ID: 413
Wang J, Villeneuve S, Zhang J, Lei PS, Miller CE, Lafaye P, Nato F, Szu SSC, Karpas A, Bystricky S, Robbins JB, Kovác P, Fournier JM, Glaudemans CPJ "On the antigenic determinants of the lipopolysaccharides of Vibrio cholerae O:1, serotypes Ogawa and Inaba" -
Journal of Biological Chemistry 273(5) (1998) 2777-2783
Monoclonal, murine IgG1s S-20-4, A-20-6, and IgA 2D6, directed against Vibrio cholerae O:1 Ogawa-lipopolysaccharide exhibited the same fine specificities and similar affinities for the synthetic methyl α-glycosides of the (oligo)saccharide fragments mimicking the Ogawa O-polysaccharide (O-PS). They did not react with the corresponding synthetic fragments of Inaba O-PS. IgG1s S-20-4 and A-20-6 have absolute affinity constants for synthetic Ogawa mono- to hexasaccharides of from approximately 10(5) to approximately 10(6) M-1. For IgG1s S-20-4, A-20-6, and IgA 2D6, the nonreducing terminal residue of Ogawa O-PS is the dominant determinant, accounting for approximately 90% of the maximal binding energy shown by these antibodies. Binding studies of derivatives of the Ogawa monosaccharide and IgGs S-20-4 and A-20-6 revealed that the C-2 O-methyl group fits into a somewhat flexible antibody cavity and that hydrogen bonds involving the oxygen and, respectively, the OH at the 2- and 3-position of the sugar moiety as well as the 2'-position in the amide side chain are required. Monoclonal IgA ZAC-3 and IgG3 I-24-2 are specific for V. cholerae O:1 serotypes Ogawa/Inaba-LPS.1 The former did not show binding with members of either series of the synthetic ligands related to the O-antigens of the Ogawa or Inaba serotypes, in agreement with its reported specificity for the lipid/core region (1). Inhibition studies revealed that the binding of purified IgG3 I-24-2 to Ogawa-LPS might be mediated by a region in the junction of the OPS to the lipid-core region of the LPS. cDNA cloning and analysis of the anti-Ogawa antibodies S-20-4, A-20-6, and 2D6 revealed a very high degree of homology among the heavy chains. Among the light chains, no such homology between S-20-4 and A-20-6 on the one hand, and 2D6 on the other hand, exists. For the anti-Inaba/Ogawa antibodies I-24-2 and ZAC-3, their heavy chains are completely different, with some homology among the light chains
Lipopolysaccharide, lipopolysaccharides, LPS, serotype, antigenic determinant, Vibrio, antigenic, determinant, Vibrio cholerae, Serotypes
NCBI PubMed ID: 9446585Journal NLM ID: 2985121RPublisher: Baltimore, MD: American Society for Biochemistry and Molecular Biology
Correspondence: glau@helix.nih.gov
Institutions: Laboratory of Medicinal Chemistry, NIDDK and the Laboratory of Developmental and Molecular Immunity, NICHD, National Institutes of Health, Bethesda, MD, USA, Unite du Cholera et des Vibrions, Centre National de Reference des Vibrions et du Cholera and Hybridolab, Institut Pasteur, Paris, France
Methods: 1H NMR, PCR, DNA sequencing, SDS-PAGE, ELISA, genetic methods
- Article ID: 925
Lei PS, Ogawa Y, Kovác P "Synthesis of the methyl a-glycoside of a trisaccharide mimicking the terminus of the antigen of Vibrio cholerae O:1, serotype Inaba" -
Carbohydrate Research 279 (1995) 117-131
Coupling of methyl 4-amino-4,6-dideoxy-2-O-4-methoxybenzyl-α-D-mannopyranoside, obtained from the corresponding 4-azido derivative by treatment with H2S, with 3-deoxy-L-glycero-tetronolactone gave the crystalline methyl 4-(3-deoxy-L-glycero-tetronamido)-4,6-dideoxy-2-O-4-methoxybenzyl-α-D-mannopyranoside (7). Subsequent acetylation of 7, followed by O-demethoxybenzylation of the 8 formed gave the crystalline methyl 3-O-acetyl-4,6-dideoxy-4-(2,4-di-O-acetyl-3-deoxy-L-glycero-tetronamido)-α-D-mannopyranoside (9), which was used as the key intermediate in the construction of the title trisaccharide. To make a glycosyl donor allowing the extension of the oligosaccharide chain at O-2, compound 9 was converted, via conventional transformations, into 3-O-acetyl-2-O-bromoacetyl-4,6-dideoxy-4-(2,4-di-O-acetyl-3-deoxy-L-glyc ero- tetronamido)-α-D-mannopyranosyl chloride (12). Condensation of 12 with 9 afforded the disaccharide 20 having a selectively removable protecting group at O-2(2). The latter was O-debromoacetylated, and the disaccharide nucleophile thus obtained was treated with 2,3-di-O-acetyl-4,6-dideoxy-4-(2,4-di-O-acetyl-3-deoxy-L-glycero-tetr onamido)-α-D-mannopyranosyl chloride to give, after O-deacetylation, the target, title trisaccharide. The constituent monosaccharide of the O-specific polysaccharide antigen of Vibrio cholerae serotype Inaba, 4-(3-deoxy-L-glycero-tetronamido)-4,6-dideoxy-D-mannopyranose (18), was obtained from the peracetate of its methyl α-glycoside by acetolysis, followed by O-deacetylation. The amorphous compound 18 was characterized by 1H and 13C NMR spectroscopy and through its crystalline α-per-O-acetyl derivative.
synthesis, antigen, serotype, trisaccharide, Vibrio, Vibrio cholerae, Vibrio cholerae O1, 4-amino-4, 6-dideoxy-D-mannose, glycoside, methyl, terminus
NCBI PubMed ID: 8593618Journal NLM ID: 0043535Publisher: Elsevier
Institutions: NIDDK, National Institutes of Health, Bethesda, MD, USA
- Article ID: 1324
Zhang J, Kovác P "Synthesis of methyl a-glycosides of some higher oligosaccharide fragments of the O-antigen of Vibrio cholerae O1, serotype Inaba and Ogawa" -
Carbohydrate Research 300(4) (1997) 329-339
The title oligosaccharides, the tri- through the hexasaccharide in the Inaba series and the penta- and the hexasaccharide in the Ogawa series, have been synthesized using 1-thioglycosides of precursors to 3-O-benzyl-perosamine (4-amino-4,6-dideoxy-D-mannose) as building blocks and N-iodosuccinimide/silver triflate as a promoter. The azido groups in the assembled oligosaccharides were reduced to amino groups, which were then acylated using 2,4-O-benzylidene-3-deoxy-L-glycero-tetronic acid as the derivatizing reagent. Catalytic hydrogenolysis, simultaneously of the benzyl and benzylidene groups, gave the desired products that were characterized by 1H and 13C NMR spectroscopy
synthesis, oligosaccharide, serotype, O-antigen, O antigen, fragment, Vibrio, Vibrio cholerae, Vibrio cholerae O1, methyl, synthetic oligosaccharides, N-iodosuccinimide silver triflate-, promoted glycosylation, tetronic acid
NCBI PubMed ID: 9210300Journal NLM ID: 0043535Publisher: Elsevier
Institutions: NIDDK, National Institutes of Health, Bethesda, MD, USA
- Article ID: 1397
Bystricky S, Szu SC, Zhang J, Kovác P "Conformational differences among mono- and oligosaccharide fragments of the O-specific polysaccharides of Vibrio cholerae O1 revealed by circular dichroism" -
Carbohydrate Research 314(1-2) (1998) 135-139
The circular dichroism (CD) of synthetic mono- and oligosaccharides that represent the terminal, non-reducing group of O-antigens of Vibrio cholerae O1 from the subtypes Ogawa and Inaba was measured in various solvents. We found differences in the CD of the monosaccharides of these subtypes that decrease with increasing chain lengths of the oligosaccharides. The differences can be explained by different orientations of the N-acyl side chain of the terminal monosaccharides. The linear relationship of ellipticity versus the number of residues in an oligosaccharide chain follows the principle of optical superposition. This, together with a similar contribution by internal units to the overall ellipticity, suggests an identical, regular conformation of oligosaccharide fragments of both Ogawa and Inaba series.
oligosaccharide, polysaccharide, conformational, O-antigens, O-specific, O-specific polysaccharide, polysaccharides, difference, fragment, Vibrio, Vibrio cholerae, Vibrio cholerae O1, O-specific polysaccharides, circular dichroism
NCBI PubMed ID: 10230041Journal NLM ID: 0043535Publisher: Elsevier
Institutions: Institute of Chemistry, Slovak Academy of Sciences, Bratislava, Slovak Republic
Methods: CD
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11. Compound ID: 870
|
S-2,4HOBut-(1-4)-+
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S-2,4HOBut-(1-4)-+ |
| |
S-2,4HOBut-(1-4)-a-D-Rhap4N2Me-(1-2)-a-D-Rhap4N-(1-2)-a-D-Rhap4N-(1-1)-Me |
Show graphically |
Structure type: oligomer
Contained glycoepitopes: IEDB_130664,IEDB_161054
The structure is contained in the following publication(s):
- Article ID: 242
Gonzalez L, Asensio JL, Ariosa-Alvarez A, Verez-Bencomo V, Jiménez-Barbero J "Solution conformation and dynamics of the trisaccharide fragments of the O-antigen of Vibrio cholerae O1, serotypes Inaba and Ogawa" -
Carbohydrate Research 321(1-2) (1999) 88-95
The conformational behavior of the trisaccharide fragments of the Ogawa and Inaba Vibrio cholerae serotypes has been studied using NMR and molecular dynamics (MD). The obtained results indicate that there are no significant differences in the major conformation and in the extent of motion of the glycosidic torsions of these molecules. The differences in biological activity are probably not due to conformational effects but to van der Waals and/or hydrogen bonding interactions between the antigens and the biological receptor.
NMR spectroscopy, Vibrio cholerae, Serotypes, Molecular mechanics
NCBI PubMed ID: 10612004Journal NLM ID: 0043535Publisher: Elsevier
Institutions: Departamento Quimica Organica Biologica, Instituto Quimica Organica, CSIC, Madrid, Spain
Methods: NMR
- Article ID: 413
Wang J, Villeneuve S, Zhang J, Lei PS, Miller CE, Lafaye P, Nato F, Szu SSC, Karpas A, Bystricky S, Robbins JB, Kovác P, Fournier JM, Glaudemans CPJ "On the antigenic determinants of the lipopolysaccharides of Vibrio cholerae O:1, serotypes Ogawa and Inaba" -
Journal of Biological Chemistry 273(5) (1998) 2777-2783
Monoclonal, murine IgG1s S-20-4, A-20-6, and IgA 2D6, directed against Vibrio cholerae O:1 Ogawa-lipopolysaccharide exhibited the same fine specificities and similar affinities for the synthetic methyl α-glycosides of the (oligo)saccharide fragments mimicking the Ogawa O-polysaccharide (O-PS). They did not react with the corresponding synthetic fragments of Inaba O-PS. IgG1s S-20-4 and A-20-6 have absolute affinity constants for synthetic Ogawa mono- to hexasaccharides of from approximately 10(5) to approximately 10(6) M-1. For IgG1s S-20-4, A-20-6, and IgA 2D6, the nonreducing terminal residue of Ogawa O-PS is the dominant determinant, accounting for approximately 90% of the maximal binding energy shown by these antibodies. Binding studies of derivatives of the Ogawa monosaccharide and IgGs S-20-4 and A-20-6 revealed that the C-2 O-methyl group fits into a somewhat flexible antibody cavity and that hydrogen bonds involving the oxygen and, respectively, the OH at the 2- and 3-position of the sugar moiety as well as the 2'-position in the amide side chain are required. Monoclonal IgA ZAC-3 and IgG3 I-24-2 are specific for V. cholerae O:1 serotypes Ogawa/Inaba-LPS.1 The former did not show binding with members of either series of the synthetic ligands related to the O-antigens of the Ogawa or Inaba serotypes, in agreement with its reported specificity for the lipid/core region (1). Inhibition studies revealed that the binding of purified IgG3 I-24-2 to Ogawa-LPS might be mediated by a region in the junction of the OPS to the lipid-core region of the LPS. cDNA cloning and analysis of the anti-Ogawa antibodies S-20-4, A-20-6, and 2D6 revealed a very high degree of homology among the heavy chains. Among the light chains, no such homology between S-20-4 and A-20-6 on the one hand, and 2D6 on the other hand, exists. For the anti-Inaba/Ogawa antibodies I-24-2 and ZAC-3, their heavy chains are completely different, with some homology among the light chains
Lipopolysaccharide, lipopolysaccharides, LPS, serotype, antigenic determinant, Vibrio, antigenic, determinant, Vibrio cholerae, Serotypes
NCBI PubMed ID: 9446585Journal NLM ID: 2985121RPublisher: Baltimore, MD: American Society for Biochemistry and Molecular Biology
Correspondence: glau@helix.nih.gov
Institutions: Laboratory of Medicinal Chemistry, NIDDK and the Laboratory of Developmental and Molecular Immunity, NICHD, National Institutes of Health, Bethesda, MD, USA, Unite du Cholera et des Vibrions, Centre National de Reference des Vibrions et du Cholera and Hybridolab, Institut Pasteur, Paris, France
Methods: 1H NMR, PCR, DNA sequencing, SDS-PAGE, ELISA, genetic methods
- Article ID: 926
Lei PS, Ogawa Y, Kovác P "Synthesis of the methyl a-glycosides of a di-, tri-, and a tetra-saccharide fragment mimicking the terminus of the O-polysaccharide of Vibrio cholerae O:1, serotype Ogawa" -
Carbohydrate Research 281 (1996) 47-60
Methyl 4-(3-deoxy-L-glycero-tetronamido)-4,6-dideoxy-2-O-methyl-α-D-mannopyranoside was acetylated, and the fully protected methyl glycoside was treated with dichloromethyl methyl ether-ZnCl2 (DCMME-ZnCl2) reagent to give 3-O-acetyl-4-(2,4-di-O-acetyl-3- deoxy-L-glycero-tetronamido)-4,6-dideoxy-2-O-methyl-α-D-mannop yranosyl chloride (3). Condensation of 3 with methyl 3-O-acetyl-4-(2,4-di-O-acetyl-3-deoxy-L-glycero-tetronamido)-4,6- dideoxy-α-D-mannopyranoside (4) gave the fully acetylated disaccharide 5, which was deacetylated yielding the methyl α-glycoside of title disaccharide. The disaccharide glycosyl donor required for the blockwise synthesis of the title tri- and the tetra-saccharide, 3-O-acetyl-4-(2,4-di-O-acetyl-3-deoxy-L-glycero-tetronamido)-4,6-d ideoxy-2-O- methyl-α-D-mannopyranosyl-(1→2)-3-O-acetyl-4- (2,4-di-O-acetyl-3-deoxy-L-glycero-tetronamido)-4,6-dideoxy-α-D-mannopyranosyl chloride (12), was obtained by condensation of 3 with the 1-O-acetyl analog of 4, followed by treatment of the disaccharide formed with DCMME-ZnCl2. The synthesis of the methyl α-glycoside of the title trisaccharide involved a condensation of 12 with 4, followed by deacetylation. Similarly, the condensation of 12 with 15, the latter being the analog of 5 having a free HO-2, followed by deacetylation, gave the methyl α-glycoside of the title tetrasaccharide. All glycosylation reactions were mediated by silver trifluoromethanesulfonate in the presence of 2,4,6-trimethylpyridine. 4-(3-Deoxy-L-glycero-tetronamido)-4,6-dideoxy-2-O-methyl-α,β-D-mannopyranose was prepared for the first time. It was characterized by NMR spectroscopy, and via its crystalline per-O-acetyl derivative. It is the saccharide whose alpha-form constitutes the terminal, non-reducing end-group of the O-PS of V. cholerea O:1, serotype Ogawa.
synthesis, tetrasaccharide, polysaccharide, serotype, Oligosaccharides, O-polysaccharide, O polysaccharide, fragment, Vibrio, Vibrio cholerae, glycoside, methyl, terminus, TRIS
NCBI PubMed ID: 8839176Journal NLM ID: 0043535Publisher: Elsevier
Institutions: NIDDK, National Institutes of Health, Bethesda, MD, USA
- Article ID: 1397
Bystricky S, Szu SC, Zhang J, Kovác P "Conformational differences among mono- and oligosaccharide fragments of the O-specific polysaccharides of Vibrio cholerae O1 revealed by circular dichroism" -
Carbohydrate Research 314(1-2) (1998) 135-139
The circular dichroism (CD) of synthetic mono- and oligosaccharides that represent the terminal, non-reducing group of O-antigens of Vibrio cholerae O1 from the subtypes Ogawa and Inaba was measured in various solvents. We found differences in the CD of the monosaccharides of these subtypes that decrease with increasing chain lengths of the oligosaccharides. The differences can be explained by different orientations of the N-acyl side chain of the terminal monosaccharides. The linear relationship of ellipticity versus the number of residues in an oligosaccharide chain follows the principle of optical superposition. This, together with a similar contribution by internal units to the overall ellipticity, suggests an identical, regular conformation of oligosaccharide fragments of both Ogawa and Inaba series.
oligosaccharide, polysaccharide, conformational, O-antigens, O-specific, O-specific polysaccharide, polysaccharides, difference, fragment, Vibrio, Vibrio cholerae, Vibrio cholerae O1, O-specific polysaccharides, circular dichroism
NCBI PubMed ID: 10230041Journal NLM ID: 0043535Publisher: Elsevier
Institutions: Institute of Chemistry, Slovak Academy of Sciences, Bratislava, Slovak Republic
Methods: CD
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12. Compound ID: 905
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S-2,4HOBut-(1-4)-+
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S-2,4HOBut-(1-4)-a-D-Rhap4N2Me-(1-2)-a-D-Rhap4N-(1--/lipid A/ |
Show graphically |
Structure type: oligomer
Aglycon: lipid A
Compound class: LPS
Contained glycoepitopes: IEDB_130664
The structure is contained in the following publication(s):
- Article ID: 259
Hisatsune K, Kondo S, Iguchi T, Ito T, Hiramatsu K "Lipopolysaccharides of Escherichia coli K12 strains that express cloned genes for the Ogawa and Inaba antigens of Vibrio cholerae O1: identification of O-antigenic factors" -
Microbiology and Immunology 40(9) (1996) 621-626
Structural and serological studies were performed with the lipopolysaccharide (LPS) expressed by Escherichia coli K12 strains No. 30 and No. 64, into which cosmid clones derived from Vibrio cholerae O1 NIH 41 (Ogawa) and NIH 35A3 (Inaba) had been introduced, respectively. The two recombinant strains, No. 30 (Ogawa) and No. 64 (Inaba), produced LPS that included, in common, the O-polysaccharide chain composed of an α(1→2)-linked N-(3-deoxy-L-glycero-tetronyl)-D-perosamine (4-amino-4,6-dideoxy-D-manno-pyranose) homopolymer attached to the core oligosaccharide of the LPS of E. coli K12. Structural analysis revealed the presence of N-(3-deoxy-L-glycero-tetronyl)-2-O-methyl-D-perosamine at the non-reducing terminus of the O-polysaccharide chain of LPS from No. 30 (Ogawa) but not from No. 64 (Inaba). Serological analysis revealed that No. 30 (Ogawa) and No. 64 (Inaba) LPS were found to share the group antigen factor A of V. cholerae O1. They were distinguished by presence of the Ogawa antigen factor B [co-existing with relatively small amounts of the Inaba antigen factor (c)] in the former LPS and the Inaba antigen factor C in the latter LPS. It appears, therefore, that No. 30 (Ogawa) and No. 64 (Inaba) have O-antigenic structures that are fully consistent with the AB(c) structure for the Ogawa and the AC structure for the Inaba O-forms of V. cholerae O1, respectively. Thus, the present study clearly confirmed our previous finding that the Ogawa antigenic factor B is substantially related to the 2-O-methyl group at the non-reducing terminus of the α(1→2)-linked N-(3-deoxy-L-glycero-tetronyl)-D-perosamine homopolymer that forms the O-polysaccharide chain of LPS of V. cholerae O1 (Ogawa).
antigen, lipopolysaccharides, Escherichia coli, O-antigenic, Vibrio cholerae O1
NCBI PubMed ID: 8908606Journal NLM ID: 7703966Publisher: Japanese Society For Bacteriology
Institutions: Department of Microbiology, School of Pharmaceutical Sciences, Josai University, Sakado, Saitama 350-02, Japan, Departament of Bacteriology, Shool of Medicine, Juntendo University, Bunkyo-ku, Tolyo 113, Japan., Departament of Bacteriology, Shool of Medicine, Juntendo University, Bunkyo-ku, Tolyo 113, Japan
Methods: methylation
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13. Compound ID: 931
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3HOPro-(1-4)-+
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3HOPro-(1-4)-a-D-Rhap4N2Me-(1-2)-a-D-Rhap4N-(1--/O-antigen/ |
Show graphically |
Structure type: oligomer
Aglycon: O-antigen
Compound class: LPS
The structure is contained in the following publication(s):
- Article ID: 268
Isshiki Y, Kondo S, Haishima Y, Iguchi T, Hisatsune K "Identification of N-3-hydroxypropionyl-2-O-methyl-D-perosamine as a specific constituent of the lipopolysaccharide from Vibrio bio-serogroup 1875 which has Ogawa antigen factor B of Vibrio cholerae O1" -
Journal of Endotoxin Research 3 (1996) 143-149
A marine vibrio, designated Vibrio bio-serogroup 1875 Original (1875 Original), has both the Ogawa antigen factor B and the Inaba antigen factor C of Vibrio cholerae O1, in addition to its own O antigen factor. Its variant strain (1875 Variant) has the Inaba antigen factor C. The O polysaccharide chain (O-chain) of the lipopolysaccharide (LPS) isolated from this vibrio has been shown to consist of an a (1-2)-linked homopolymer of /V-3-hydroxypropionyl-D-perosamine. The present structural study revealed the presence of N-3-hydroxypropionyl-2-O-methyl-D-perosamine at the non-reducing terminus of the O-chain from the LPS of 1875 Original but not from that of the 1875 Variant. Since 2-O-methyi-D-perosamine is known as an Ogawa-specific constituent of the O-chain from V. cholerae O1 Ogawa O form, it appears that the presence of 2-O-methyl-D-perosamine at the non-reducing terminus of the a (1-2)-linked D-perosamine homopolymer that forms the O-chain is responsible for the expression of the serological specificity of the Ogawa antigen factor B of 1875 Original.
Lipopolysaccharide, antigen, Vibrio, Vibrio cholerae O1, 3-hydroxypropionyl, 4-amino-4, 6-dideoxy-D-mannose, factor B
Publication DOI: 10.1177/096805199600300207Journal NLM ID: 9433350Publisher: Maney Publishing
Institutions: Department of Microbiology, School of Pharmaceutical Sciences, Josai University, Saitama, Japan, Division of Microbiology, National Institute of Hygienic Sciences, Tokyo, Japan
Methods: 13C NMR, 1H NMR, methylation
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14. Compound ID: 975
Structure type: homopolymer
Trivial name: perosamine
Compound class: O-polysaccharide, O-antigen, LPS
The structure is contained in the following publication(s):
- Article ID: 269
Isshiki Y, Haishima Y, Kondo S, Hisatsune K "Immunochemistry of group A and Inaba C antigen factors constituting the O antigen of O1 Vibrio cholerae" -
European Journal of Biochemistry 229 (1995) 583-588
Serological cross-reactivity among intact lipopolysaccharides (LPS) from O1 Vibrio cholerae Inaba O-form (Inaba), Yersinia enterocolitica O9 (O9), non-O1 V. cholerae serogroup Hakata (Hakata) and Vibrio bio-serogroup 1875 Variant (1875 Variant) (all of which share Inaba antigen factor C), as well as a total of six kinds of chemically modified LPS (three from O9 and three from Inaba) was demonstrated by passive hemolysis and passive hemolysis inhibition by using these LPS as antigen for sensitizing sheep red blood cells and as inhibitor. These intact as well as chemically modified LPS contained, in their O polysaccharide chain, α(1→2)-linked linear perosamine (4-amino-4,6-dideoxy-D-manno-pyranose) homopolymers with different N-acyl groups: their acyl groups comprise 3-deoxy-L-glycero-tetronyl (Inaba LPS), formyl (O9 LPS), 3-hydroxypropionyl (1875 Variant LPS), acetyl (Hakata LPS and artificially introduced into Inaba and O9 LPS), propionyl and butyryl (both artificially introduced into Inaba and O9 LPS) groups. N-Deacylation of the α(l→2)-linked N-(3-deoxy-L-glycero-tetronyl)perosamine homopolymer of Inaba and the N-formyl one of O9 LPS resulted in virtual elimination of their serological reactivity with both homologous and heterologous antisera. Furthermore, when the resultant NH2 groups of the N-deacylated perosamine homopolymers of both LPS were N-acylated with acetyl, propionyl or butyryl groups, they markedly recovered both of their serological reactivities. These results are compatible with the interpretation that the Inaba antigen factor C possessed by the four bacteria is substantially related to the common presence of N-acyl groups, regardless of their identity, residing in the perosamine residues constituting the O polysaccharide chain of their LPS. It was also indicated that the group antigen factor A of O1 V. cholerae is substantially related to the 3-deoxy-L-glycerotetronyl groups residing in the perosamine homopolymer of Inaba LPS.
antigen, LPS, O-antigen, Vibrio, Vibrio cholerae O1
NCBI PubMed ID: 7538078Publication DOI: 10.1111/j.1432-1033.1995.0583k.xJournal NLM ID: 0107600Publisher: Oxford, UK: Blackwell Science Ltd. on behalf of the Federation of European Biochemical Societies
Institutions: Department of Microbiology, School of Pharmaceutical Sciences Josai University, Saitama, Japan, Division of Microbiology, National Institute of Hygienic Science, Tokyo, Japan
Methods: 13C NMR, 1H NMR, methylation, gel filtration, GC-MS, GLC, serological methods
- Article ID: 534
Ovchinnikova OG, Kocharova NA, Katzenellenbogen E, Zatonsky GV, Shashkov AS, Knirel YA, Lipinski T, Gamian A "Structures of two O-polysaccharides of the lipopolysaccharide of Citrobacter youngae PCM 1538 (serogroup O9)" -
Carbohydrate Research 339(4) (2004) 881-884
Mild acid degradation of the lipopolysaccharide of Citrobacter youngae O9, strain PCM 1538 released a homopolysaccharide of 4-acetamido-4,6-dideoxy-D-mannose (D-Rha4NAc, N-acetyl-D-perosamine). Studies by methylation analysis and (1)H and (13)C NMR spectroscopy, using two-dimensional (1)H,(1)H COSY, TOCSY, NOESY and H-detected (1)H,(13)C HSQC experiments showed the presence of two structurally different polysaccharides consisting of the following units: →2)-α-D-Rhap4NAc-(1→ and →3)-α-D-Rhap4NAc-(1→3)-β-D-Rhap4NAc-(1→.
Lipopolysaccharide, O-antigen, polysaccharide structure, 6-dideoxy-D-mannose, Citrobacter youngae, 4-Acetamido-4
NCBI PubMed ID: 14980832Publication DOI: 10.1016/j.carres.2003.12.026Journal NLM ID: 0043535Publisher: Elsevier
Correspondence: knirel@ioc.ac.ru
Institutions: N.D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia, L. Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wroclaw, Poland
Methods: methylation, NMR-2D, NMR, sugar analysis
- Article ID: 945
Lipinski T, Zatonsky GV, Kocharova NA, Jaquinod M, Forest E, Shashkov AS, Gamian A, Knirel YA "Structures of two O-chain polysaccharides of Citrobacter freundii O9a,9b lipopolysaccharide. A new homopolymer of 4-amino-4,6-dideoxy-D-mannose (perosamine)" -
European Journal of Biochemistry 269(1) (2002) 93-99
Mild acid degradation of the lipopolysaccharide of Citrobacter gillenii O9a,9b released a polysaccharide (PS), which was found to consist of a single monosaccharide, 4-acetamido-4,6-dideoxy-D-mannose (D-Rha4NAc, N-acetyl-D-perosamine). PS was studied by methylation analysis and 1H NMR and 13C NMR spectroscopy, using two-dimensional 1H,1H COSY, TOCSY, NOESY, and H-detected 1H,13C heteronuclear correlation experiments. It was found that PS includes two structurally different polysaccharides: an alfa 1→2-linked homopolymer of N-acetyl-D-perosamine [ →2)-a-D-Rhap4NAc-(1→, PS2] and a polysaccharide composed of tetrasaccharide repeating units (PS1) with the following structure: →3)-a-D-Rhap4NAc-(1→2)-a-D-Rhap4NAc-(1→2)-a-D-Rhap4NAc-(1→3)-a-D-Rhap4NAc2Ac-(1→ where the degree of O-acetylation of a 3-substituted Rha4NAc residue at position 2 is ~70%. PS could be fractionated into PS1 and PS2 by gel-permeation chromatography on TSK HW-50S. Matrix-assisted laser desorption ionization MS data indicate sequential chain elongation of both PS1 and PS2 by a single sugar unit, with O-acetylation in PS1 beginning at a certain chain length. Anti-(C. gillenii O9a,9b) serum reacted with PS1 in double immunodiffusion and immunoblotting, whereas neither PS2 nor the lipopolysaccharide of Vibrio cholerae O1 with a structurally related O-chain polysaccharide were reactive.
Lipopolysaccharide, LPS, structure, polysaccharide, O-antigen, repeating unit, O-specific, O-specific polysaccharide, polysaccharides, 6-dideoxy-d-glucose, O-chain, pentasaccharide, O-specific polysaccharides, 4-amino-4, 6-dideoxy-D-mannose, homopolymer, perosamine, Citrobacter, Citrobacter freundii, 4-Acetamido-4
Journal NLM ID: 0107600Publisher: Oxford, UK: Blackwell Science Ltd. on behalf of the Federation of European Biochemical Societies
Correspondence: gamian@immuno.iitd.pan.wroc.pl
Institutions: N.D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia, L. Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wroclaw, Poland, CNRS and CEA, Institut de Biologie Structurale, LSMP, Grenoble, France
Methods: methylation, NMR-2D, NMR, sugar analysis, MS, de-O-acetylation
- Article ID: 1468
Knirel YA, Kocharova NA, Bystrova OV, Katzenellenbogen E, Gamian A "Structures and serology of the O-specific polysaccharides of bacteria of the genus Citrobacter" -
Archivum Immunologiae et Therapiae Experimentalis 50(6) (2002) 379-391
The review presents the structures of the O-specific polysaccharides (O-antigens) of the lipopolysaccharides isolated from over 25 Citrobacter strains, which represent different species and serogroups. The correlation between O-antigen structure and immunospecificity as well as numerous cross-reactions between Citrobacter and other enterobacterial species are discussed.
Lipopolysaccharide, structure, O-antigen, O-specific polysaccharide, serology, Citrobacter, immunospecificity
NCBI PubMed ID: 12546064Journal NLM ID: 0114365Publisher: Basel, Boston: Birkhaüser
Correspondence: knirel@ioc.ac.ru
Institutions: N.D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia
- Article ID: 1539
Leone S, Izzo V, Lanzetta R, Molinaro A, Parrilli M, Di Donato A "The structure of the O-polysaccharide from Pseudomonas stutzeri OX1 containing two different 4-acylamido-4,6-dideoxy-residues, tomosamine and perosamine" -
Carbohydrate Research 340(4) (2005) 651-656
The structure of the O-polysaccharide from the lipopolysaccharide of Pseudomonas stutzeri OX1 was determined by chemical procedures and by 1D and 2D NMR spectroscopy. The analysis revealed the presence of a heterogeneous polymer made by 4-acetamido-4,6-dideoxy-d-mannopyranose (d-Rhap4NAc) and 4-formamido-4,6-dideoxy-d-galactopyranose (d-Fucp4NFo). The combination of chemical and NMR analyses indicates that the heterogeneity of the polymer depends on its non-stoichiometric glycosylation by Fuc4NFo, as shown below:The structure of the heterogeneous polymer was confirmed by Smith degradation that significantly simplified the structure of the O-polysaccharide, allowing for the isolation and identification of a linear homopolymer of Rhap4NAc
Lipopolysaccharide, NMR, structure, analysis, Pseudomonas, polymer, NMR spectroscopy, O-polysaccharide, O polysaccharide, chemical, identification, spectroscopy, degradation, isolation, linear, glycosylation, 2D NMR spectroscopy, 2D NMR, homopolymer, perosamine, Smith degradation, heterogeneity, Pseudomonas stutzeri
NCBI PubMed ID: 15721336Journal NLM ID: 0043535Publisher: Elsevier
Institutions: Dipartimento di Chimica Organica e Biochimica, Universita degli Studi di Napoli Federico II, Via Cintia, 4 I-80126 Napoli, Italy
Methods: methylation, NMR, Smith degradation, composition analysis
- 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: 2748
Haishima Y, Kondo S, Hisatsune K "The occurrence of a(1-2) linked N-acetylperosamine-homopolymer in lipopolysaccharides of Non-O1 Vibrio cholerae possessing an antigenic factor in common with O1 V. cholerae" -
Microbiology and Immunology 34 (1990) 1049-1054
Chemical analysis was carried out on lipopolysaccharides from Vibrio cholerae bio-serogroup Hakata 487-85. The O-specific chain of the phenol-soluble lipopolysaccharides was demonstrated by 13C-NMR spectroscopy and methylation analysis to contain a linear homopolymer of α(1→2) linked N-acetylperosamine (4-acetamido-4,6-dideoxy-D-mannopyranose), which was closely similar to but not identical to a linear α(1→2) linked N-3-deoxy-L-glycerotetronyl (S-2,4-dihydroxybutyryl) perosamine-homopolymer constituting that of O1 Vibrio cholerae lipopolysaccharides.
NCBI PubMed ID: 2098633Journal NLM ID: 7703966Publisher: Japanese Society For Bacteriology
Institutions: Department of Microbiology, School of Pharmaceutical Sciences, Josai University, Saitama
- 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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15. Compound ID: 976
Structure type: homopolymer
Trivial name: perosamine homopolymer derivative
Compound class: O-polysaccharide, O-antigen
The structure is contained in the following publication(s):
- Article ID: 269
Isshiki Y, Haishima Y, Kondo S, Hisatsune K "Immunochemistry of group A and Inaba C antigen factors constituting the O antigen of O1 Vibrio cholerae" -
European Journal of Biochemistry 229 (1995) 583-588
Serological cross-reactivity among intact lipopolysaccharides (LPS) from O1 Vibrio cholerae Inaba O-form (Inaba), Yersinia enterocolitica O9 (O9), non-O1 V. cholerae serogroup Hakata (Hakata) and Vibrio bio-serogroup 1875 Variant (1875 Variant) (all of which share Inaba antigen factor C), as well as a total of six kinds of chemically modified LPS (three from O9 and three from Inaba) was demonstrated by passive hemolysis and passive hemolysis inhibition by using these LPS as antigen for sensitizing sheep red blood cells and as inhibitor. These intact as well as chemically modified LPS contained, in their O polysaccharide chain, α(1→2)-linked linear perosamine (4-amino-4,6-dideoxy-D-manno-pyranose) homopolymers with different N-acyl groups: their acyl groups comprise 3-deoxy-L-glycero-tetronyl (Inaba LPS), formyl (O9 LPS), 3-hydroxypropionyl (1875 Variant LPS), acetyl (Hakata LPS and artificially introduced into Inaba and O9 LPS), propionyl and butyryl (both artificially introduced into Inaba and O9 LPS) groups. N-Deacylation of the α(l→2)-linked N-(3-deoxy-L-glycero-tetronyl)perosamine homopolymer of Inaba and the N-formyl one of O9 LPS resulted in virtual elimination of their serological reactivity with both homologous and heterologous antisera. Furthermore, when the resultant NH2 groups of the N-deacylated perosamine homopolymers of both LPS were N-acylated with acetyl, propionyl or butyryl groups, they markedly recovered both of their serological reactivities. These results are compatible with the interpretation that the Inaba antigen factor C possessed by the four bacteria is substantially related to the common presence of N-acyl groups, regardless of their identity, residing in the perosamine residues constituting the O polysaccharide chain of their LPS. It was also indicated that the group antigen factor A of O1 V. cholerae is substantially related to the 3-deoxy-L-glycerotetronyl groups residing in the perosamine homopolymer of Inaba LPS.
antigen, LPS, O-antigen, Vibrio, Vibrio cholerae O1
NCBI PubMed ID: 7538078Publication DOI: 10.1111/j.1432-1033.1995.0583k.xJournal NLM ID: 0107600Publisher: Oxford, UK: Blackwell Science Ltd. on behalf of the Federation of European Biochemical Societies
Institutions: Department of Microbiology, School of Pharmaceutical Sciences Josai University, Saitama, Japan, Division of Microbiology, National Institute of Hygienic Science, Tokyo, Japan
Methods: 13C NMR, 1H NMR, methylation, gel filtration, GC-MS, GLC, serological methods
- Article ID: 297
Kondo S, Haishima Y, Ishida K, Isshiki Y, Hisatsune K "The O-polysaccharide of lipopolysaccharide isolated from Vibrio fluvialis O19 is identical to that of Vibrio bioserogroup 1875 variant" -
Microbiology and Immunology 44(11) (2000) 941-944
A structural analysis has been carried out on the O-polysaccharide of lipopolysaccharide (LPS) isolated from Vibrio fluvialis 181-86 (Kobe) serotype O19 (O19) which has the Inaba antigen factor C of O1 V. cholerae and factors D and E in common with Vibrio bioserogroup 1875. The O-polysaccharide of O19 was characterized as an α(1→2)-linked homopolymer of N-3-hydroxypropionyl-D-perosamine (4-amino-4,6-dideoxy-D-mannopyranose), which was identical to that of Vibrio bioserogroup 1875 Variant. Passive hemolysis and passive hemolysis inhibition analysis performed using anti-factor D, E and anti-factor E antisera, demonstrated that the LPS from O19 harbored O-antigenic factors identical to those of the LPS from Vibrio bioserogroup 1875 Variant.
Lipopolysaccharide, O-polysaccharide, O polysaccharide, Vibrio, variant, Vibrio fluvialis, Vibrio fluvialis O19
NCBI PubMed ID: 11145275Journal NLM ID: 7703966Publisher: Japanese Society For Bacteriology
Correspondence: kondo@josai.ac.jp
Institutions: Department of Microbiology, School of Pharmaceutical Sciences, Josai University, Sakado, Saitama, Japan.
- Article ID: 1735
Kondo S, Ishida K, Isshiki Y, Haishima Y, Iguchi T, Hisatsune K "N-3-Hydroxypropionyl-a-D-perosamine homopolymer constituting the O-chain of lipopolysaccharides from Vibrio bioserogroup 1875 possessing antigenic factor(s) in common with O1 Vibrio cholerae" -
Biochemical Journal 292 (1993) 531-535
A structural study was performed by 13C-n.m.r. spectroscopy and methylation analysis of the O-chain of lipopolysaccharide (LPS) from Vibrio bioserogroup 1875 possessing antigenic factor(s) in common with O1 Vibrio cholerae. It was demonstrated to contain a linear homopolymer of (1→2)-linked N-3-hydroxypropionyl-α-D-perosamine [4-(3-hydroxypropanamido)-4,6-dideoxy-α-D-mannopyranose], which is very similar to, but not identical with, both (1→2)-linked linear N-3-deoxy-L-glycero-tetronyl(S-2,4-dihydroxybutyryl)-α-D-perosamine homopolymer and (1→2)-linked linear N-acetyl-α-D-perosamine homopolymer which constitute the O-chains of O1 V. cholerae and non-O1 V. cholerae bioserogroup Hakata LPS respectively.
NCBI PubMed ID: 8503886Journal NLM ID: 2984726RPublisher: London, UK : Published by Portland Press on behalf of the Biochemical Society
Institutions: Department of Microbiology, School of Pharmaceutical Sciences, Josai University, Saitama, Japan
- 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: 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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