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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

• Article ID: 240
  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: 11081580
  Journal NLM ID: 8907468
  Publisher: 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
  
   
  
  Brucella melitensis 16M
  CSDB ID 6924 (all data & tools)
• Article ID: 604
  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: 8576040
  Journal NLM ID: 2985120R
  Publisher: 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
  
   
  
  Brucella abortus 2308
  CSDB ID 221 (all data & tools)
• Article ID: 773
  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: 9801349
  Journal NLM ID: 9421292
  Publisher: 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
  
   
  
  Brucella
  CSDB ID 1327 (all data & tools)
• Article ID: 1753
  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: 3442684
  Journal NLM ID: 0370623
  Publisher: American Chemical Society
  Institutions: Division of Biological Science, National Research Council of Canada, Ottawa
  Methods: 13C NMR, 1H NMR
  
   
  
  Brucella melitensis
  CSDB ID 107325 (all data & tools)
• Article ID: 1777
  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: 7533007
  Journal NLM ID: 0376536
  Publisher: Nauka/Interperiodica
  Institutions: Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia
  
   
  
  Brucella melitensis 16M
  CSDB ID 108579 (all data & tools)
• Article ID: 2534
  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: 0246127
  Publisher: American Society for Microbiology
  Methods: 13C NMR
  
   
  
  Brucella melitensis
  CSDB ID 130218 (all data & tools)
• Article ID: 2705
  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
  
  Journal NLM ID: 0375371
  
   
  
  Brucella melitensis B115
  CSDB ID 139863 (all data & tools)
• Article ID: 2803
  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: 0376514
  WWW link: http://pac.iupac.org/publications/pac/pdf/1989/pdf/6107x1171.pdf
  Publisher: Oxford: Blackwell Scientific Publications
  Institutions: Division of Biological Science, National Research Council of Canada, Ottawa, Ontario, K1A OR6, Canada
  
   
  
  Brucella melitensis
  CSDB ID 148075 (all data & tools)
• Article ID: 4306
  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: 22475157
  Publication DOI: 10.1016/j.carres.2012.02.027
  Journal NLM ID: 0043535
  Publisher: Elsevier
  Correspondence: C. Whitfield
  Institutions: Department of Molecular and Cellular Biology, University of Guelph, Guelph, Ontario, Canada N1G 2W1
  
   
  
  Brucella melitensis 16M
  CSDB ID 28375 (all data & tools)
• Article ID: 4499
  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: 23567915
  Publication DOI: 10.1016/j.bbamem.2013.03.029
  Journal NLM ID: 0217513
  Publisher: 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
  
   
  
  Brucella melitensis
  CSDB ID 29942 (all data & tools)
• Article ID: 4559
  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: 23664729
  Publication DOI: 10.1016/j.carres.2013.03.021
  Journal NLM ID: 0043535
  Publisher: 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
  
   
  
  Brucella melitensis 16M
  CSDB ID 29248 (all data & tools)
• 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: 28924631
  Publication DOI: 10.1007/s13361-017-1805-x
  Journal NLM ID: 9010412
  Publisher: 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
  
   
  
  Brucella melitensis 16M
  CSDB ID 12061 (all data & tools)

Show legend Show as text

Structure type: polymer chemical repeating unit
Contained glycoepitopes: IEDB_130647,IEDB_131172,IEDB_134279,IEDB_134280,IEDB_134281,IEDB_1397515,IEDB_2116320,IEDB_434544,IEDB_434545,IEDB_434546

• Article ID: 4306
  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: 22475157
  Publication DOI: 10.1016/j.carres.2012.02.027
  Journal NLM ID: 0043535
  Publisher: Elsevier
  Correspondence: C. Whitfield
  Institutions: Department of Molecular and Cellular Biology, University of Guelph, Guelph, Ontario, Canada N1G 2W1
  
   
  
  Brucella melitensis 16M
  CSDB ID 28375 (all data & tools)

Show legend Show as text

Structure type: polymer chemical repeating unit
Aglycon: -2)[Fo(1-4)]aDRhap4N(1- (ID 29629)
Compound class: O-polysaccharide
Contained glycoepitopes: IEDB_130647,IEDB_131172,IEDB_134279,IEDB_134280,IEDB_1397515,IEDB_2116320,IEDB_434544,IEDB_434546

• Article ID: 4559
  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: 23664729
  Publication DOI: 10.1016/j.carres.2013.03.021
  Journal NLM ID: 0043535
  Publisher: 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
  
   
  
  Brucella melitensis 16M, Brucella abortus biotype 4, Brucella suis 4, Brucella melitensis 3
  CSDB ID 29626 (all data & tools)

Show legend Show as text

Structure type: oligomer
Compound class: O-polysaccharide
Contained glycoepitopes: IEDB_130647,IEDB_131172,IEDB_134279,IEDB_134280,IEDB_134281,IEDB_1397515,IEDB_2116320,IEDB_434544,IEDB_434546,IEDB_434547,IEDB_628715

• Article ID: 6220
  Duncombe L, Howells L, Haughey A, Taylor AV, Kaveh D, Erdenlig Gurbilek S, Dell A, Hitchen PG, Haslam SM, Mandal SS, Ganesh NV, Bundle DR, McGiven J "The Tip of Brucella O-Polysaccharide Is a Potent Epitope in Response to Brucellosis Infection and Enables Short Synthetic Antigens to Be Superior Diagnostic Reagents" - Microorganisms 10(4) (2022) 708
  

  Brucellosis is a global disease and the world's most prevalent zoonosis. All cases in livestock and most cases in humans are caused by members of the genus Brucella that possess a surface O-polysaccharide (OPS) comprised of a rare monosaccharide 4-deoxy-4-formamido-D-mannopyranose assembled with α1,2 and α1,3 linkages. The OPS of the bacterium is the basis for serodiagnostic tests for brucellosis. Bacteria that also contain the same rare monosaccharide can induce antibodies that cross-react in serological tests. In previous work we established that synthetic oligosaccharides, representing elements of the Brucella A and M polysaccharide structures, were excellent antigens to explore the antibody response in the context of infection, immunisation and cross reaction. These studies suggested the existence of antibodies that are specific to the tip of the Brucella OPS. Sera from naturally and experimentally Brucella abortus-infected cattle as well as from cattle experimentally infected with the cross-reactive bacterium Yersinia enterocolitica O:9 and field sera that cross react in conventional serological assays were studied here with an expanded panel of synthetic antigens. The addition of chemical features to synthetic antigens that block antibody binding to the tip of the OPS dramatically reduced their polyclonal antibody binding capability providing conclusive evidence that the OPS tip (non-reducing end) is a potent epitope. Selected short oligosaccharides, including those that were exclusively α1,2 linked, also demonstrated superior specificity when evaluated with cross reactive sera compared to native smooth lipopolysaccharide (sLPS) antigen and capped native OPS. This surprising discovery suggests that the OPS tip epitope, even though common to both Brucella and Y. enterocolitica O:9, has more specific diagnostic properties than the linear portion of the native antigens. This finding opens the way to the development of improved serological tests for brucellosis.

  epitope, O-polysaccharide, Brucella

  NCBI PubMed ID: 35456759
  Publication DOI: 10.3390/microorganisms10040708
  Journal NLM ID: 101625893
  Publisher: Basel, Switzerland: MDPI AG
  Correspondence: john.mcgiven@apha.gov.uk
  Institutions: Department of Chemistry, University of Alberta, Edmonton, AB, Canada, Department of Bacteriology, Animal & Plant Health Agency, OIE/FAO Brucellosis Reference Laboratory, Woodham Lane, Addlestone, Surrey KT15 3NB, UK, Microbiology Department, Faculty of Veterinary Medicine, Harran University, 63500 Sanliurfa, Turkey, Department of Life Sciences, Imperial College London, London SW7 5NH, UK
  Methods: SDS-PAGE, ELISA, mild acid hydrolysis, chemical synthesis, MALDI-TOF MS, serological methods, HPLC, extraction, antibody binding, conjugation, HPLC-ESI-MS
  
   
  
  Brucella melitensis 16M, Brucella melitensis 3, Brucella abortus biotype 4, Brucella suis 4
  CSDB ID 8451 (all data & tools)