Crystal structure to 2.45 Å resolution of a monoclonal Fab specific for the <i>Brucella</i> A cell wall polysaccharide antigen

Rose, David R.; Przybylska, M.; To, Rebecca; Kayden, C. S.; Oomen, Ray; Vorberg, Ewald; Young, N. Martin; Bundle, David R.

doi:10.1002/pro.5560020705

Cited by 58 publications

(29 citation statements)

References 37 publications

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“…Synthetic compounds mimicking the inner perosamine residues of the O-SP Ag could therefore be expected to elicit Abs specific for both the Ogawa and Inaba serotypes. Such Abs with a groove-shaped binding site were indeed observed for the Brucella A cell wall polysaccharide Ag (38), an O-SP-related molecule that is also composed of a linear chain of ␣ (1, 2)-linked perosamine units (39). Table 4.…”

Section: Resultsmentioning

confidence: 84%

Crystal structure of an anti-carbohydrate antibody directed againstVibrio choleraeO1 in complex with antigen: Molecular basis for serotype specificity

Villeneuve

Souchon

Riottot

et al. 2000

Proc. Natl. Acad. Sci. U.S.A.

105

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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 monosaccharide 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.

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Section: Resultsmentioning

confidence: 84%

Crystal structure of an anti-carbohydrate antibody directed againstVibrio choleraeO1 in complex with antigen: Molecular basis for serotype specificity

Villeneuve

Souchon

Riottot

et al. 2000

Proc. Natl. Acad. Sci. U.S.A.

105

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“…Cavity-like and groove-type sites for anti-carbohydrate antibodies, including antibodies to LPS OAg from Shigella flexneri, Vibrio cholerae, and Brucella abortus, were supported by immunochemical, computer modeling, and x-ray crystallographic studies. (41)(42)(43)(44)(45)(46)(47)(48)(49)(50) These showed a maximum of five sugar residues accommodated by cavity-type sites and six to eight sugar residues accommodated by groove-type sites. In the current study, FB11 may have a cavity-type site and the other three anti-Ft OAg MAbs likely have groove-type sites.…”

Section: Biacore Analysis Confirms Higher Avidity Of End-binding Mabmentioning

confidence: 99%

Characterization of Monoclonal Antibodies to Terminal and Internal O-Antigen Epitopes ofFrancisella tularensisLipopolysaccharide

Roche

Hui

et al. 2011

Hybridoma

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The lipopolysaccharide (LPS) of Francisella tularensis (Ft), the Gram negative bacterium that causes tularemia, has been shown to be a main protective antigen in mice and humans; we have previously demonstrated that murine anti-Ft LPS IgG2a monoclonal antibodies (MAbs) can protect mice against otherwise lethal intranasal infection with the Ft live vaccine strain (LVS). Here we show that four IgG2a anti-LPS MAbs are specific for the O-polysaccharide (O-antigen [OAg]) of Ft LPS. But whereas three of the MAbs bind to immunodominant repeating internal epitopes, one binds to a unique terminal epitope of Ft OAg. This was deduced from its even binding to both long and short chains of the LPS ladder in Western blots, its rapid decrease in ELISA binding to decreasing solid-phase LPS concentrations, its inability to compete for LPS binding with a representative of the other three MAbs, and its inability to immunoprecipitate OAg despite its superior agglutination titer. Biacore analysis showed the end-binding MAb to have higher bivalent avidity for Ft OAg than the internal-binding MAbs and provided an immunogenicity explanation for the predominance of internal-binding anti-Ft OAg MAbs. These findings demonstrate that non-overlapping epitopes can be targeted by antibodies to Ft OAg, which may inform the design of vaccines and immunotherapies against tularemia.

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“…Thus, in addition to the branched glucopyranosyl residue E behaving as an anchor and to the trisaccharide A(E)B providing the critical epitope exposed on the O-SP, chain elongation also takes part in O-SP:mIgA recognition, highlighting the essential contribution of some kind of conformational epitope or presentation in an extended surface. However, we are aware that small changes at the V L :V H interface of the mAb may result in significant alteration of the binding mode, which cannot be predicted at this stage (80). Thus, data provided here are only meant to provide a model of S. flexneri 5a O-SP binding to a homologous protective mIgA, which needs to be further assessed based on crystallographic data.…”

Section: Discussionmentioning

confidence: 98%

Toward a Better Understanding of the Basis of the Molecular Mimicry of Polysaccharide Antigens by Peptides

Clément

Fortuné

Phalipon

et al. 2006

Journal of Biological Chemistry

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Protein conjugates of oligosaccharides or peptides that mimic complex bacterial polysaccharide antigens represent alternatives to the classical polysaccharide-based conjugate vaccines developed so far. Hence, a better understanding of the molecular basis ensuring appropriate mimicry is required in order to design efficient carbohydrate mimic-based vaccines. This study focuses on the following two unrelated sets of mimics of the Shigella flexneri 5a O-specific polysaccharide (O-SP): (i) a synthetic branched pentasaccharide known to mimic the average solution conformation of S. flexneri 5a O-SP, and (ii) three nonapeptides selected upon screening of phagedisplayed peptide libraries with two protective murine monoclonal antibodies (mAbs) of the A isotype specific for S. flexneri 5a O-SP. By inducing anti-O-SP antibodies upon immunization in mice when appropriately presented to the immune system, the pentasaccharide and peptides p100c and p115, but not peptide p22, were qualified as mimotopes of the native antigen. NMR studies based on transferred NOE (trNOE) experiments revealed that both kinds of mimotopes had an average conformation when bound to the mAbs that was close to that of their free form. Most interestingly, saturation transfer difference (STD) experiments showed that the characteristic turn conformations adopted by the major conformers of p100c and p115, as well as of p22, are clearly involved in mAb binding. These latter experiments also showed that the branched glucose residue of the pentasaccharide was a key part of the determinant recognized by the protective mAbs. Finally, by using NMRderived pentasaccharide and peptide conformations coupled to STD information, models of antigen-antibody interaction were obtained. Most interestingly, only one model was found compatible with experimental data when large O-SP fragments were docked into one of the mIgA-binding sites. This newly made available system provides a new contribution to the understanding of the molecular mimicry of complex polysaccharides by peptides and short oligosaccharides.

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Crystal structure to 2.45 Å resolution of a monoclonal Fab specific for the Brucella A cell wall polysaccharide antigen

Cited by 58 publications

References 37 publications

Crystal structure of an anti-carbohydrate antibody directed againstVibrio choleraeO1 in complex with antigen: Molecular basis for serotype specificity

Crystal structure of an anti-carbohydrate antibody directed againstVibrio choleraeO1 in complex with antigen: Molecular basis for serotype specificity

Characterization of Monoclonal Antibodies to Terminal and Internal O-Antigen Epitopes ofFrancisella tularensisLipopolysaccharide

Toward a Better Understanding of the Basis of the Molecular Mimicry of Polysaccharide Antigens by Peptides

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