title: Gonococcal outer-membrane protein PIB: comparative sequence analysis and localization of epitopes which are recognized by type-specific and cross-reacting monoclonal antibodies.
abstract: Comparison of the inferred amino acid sequence of outer-membrane protein PIB from gonococcal strain P9 with those from other serovars reveals that sequence variations occur in two discrete regions of the molecule centred on residues 196 (Var1) and 237 (Var2). A series of peptides spanning the amino acid sequence of the protein were synthesized on solid-phase supports and reacted with a panel of monoclonal antibodies (mAbs) which recognize either type-specific or conserved antigenic determinants on PIB. Four type-specific mAbs reacted with overlapping peptides in Var1 between residues 192-198. Analysis of the effect of amino acid substitutions revealed that the mAb specificity is generated by differences in the effect of single amino acid changes on mAb binding, so that antigenic differences between strains are revealed by different patterns of reactivity within a panel of antibodies. The variable epitopes in Var1 recognized by the type-specific mAbs lie in a hydrophilic region of the protein exposed on the gonococcal surface, and are accessible to complement-mediated bactericidal lysis. In contrast, the epitope recognized by mAb SM198 is highly conserved but is not exposed in the native protein and the antibody is non-bactericidal. However, the conserved epitope recognized by mAb SM24 is centred on residues 198-199, close to Var1 , and is exposed for bactericidal killing.
authors.affiliation: Department of Microbiology, Southampton University Medical School, Southampton General Hospital, UK.
authors.forename: N J
authors.initials: NJ
authors.lastname: Butt
authors.forename: M
authors.initials: M
authors.lastname: Virji
authors.forename: F
authors.initials: F
authors.lastname: Vayreda
authors.forename: P R
authors.initials: PR
authors.lastname: Lambden
authors.forename: J E
authors.initials: JE
authors.lastname: Heckels
mesh_terms: D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology; D001425:Bacterial Outer Membrane Proteins / Q000032:analysis* / Q000276:immunology; D001483:Base Sequence; D003429:Cross Reactions; D004269:DNA, Bacterial / Q000737:chemistry; D000939:Epitopes; D008969:Molecular Sequence Data; D009344:Neisseria gonorrhoeae / Q000235:genetics / Q000276:immunology*; D018272:Porins
chemical_list: D000911:Antibodies, Monoclonal; D001425:Bacterial Outer Membrane Proteins; D004269:DNA, Bacterial; D000939:Epitopes; D018272:Porins; C057699:porin protein, Neisseria