title: Structural and functional properties of a 69-kilodalton outer membrane protein of Bordetella pertussis.
abstract: A-69-kDa outer membrane protein present on virulent Bordetella pertussis cells is recognized by the agglutinating monoclonal antibodies BPE3, BPD8, and BPE8. The amino acid composition of this protein, purified from heat extracts of B. pertussis BP353 cells, is different from that of the two major fimbrial antigens of B. pertussis, which is consistent with its being a nonfimbrial protein based on other criteria. Western blot analysis using the monoclonal antibody BPE3 demonstrated that a slightly larger but antigenically cross-reactive protein is also expressed by Bordetella bronchiseptica and Bordetella parapertussis. In addition, a large molecular weight species of about 180-kDa is found in outer membrane extracts of B. bronchiseptica which may represent a precursor form of the protein or indicate that the protein can exist as an oligomer. The monoclonal antibody BPD8 directed against the 69-kDa protein almost completely inhibited the enzymatic activity of adenylate cyclase purified from B. pertussis and also inhibited the intoxication of mammalian cells by this enzyme. Since little enzymatic activity was found associated with the purified 69-kDa protein, these data suggest a role for the 69-kDa protein in regulating the adenylate cyclase toxin of B. pertussis. An additional monoclonal antibody directed against the 69-kDa protein, BPE8, decreases lymphocytosis and delays death in mice receiving a respiratory challenge of virulent B. pertussis cells. These studies suggest that further investigation into the role of this protein as a protective antigen and vaccine candidate is warranted.
authors.affiliation: Division of Bacterial Products, Food and Drug Administration, Bethesda, MD 20892.
authors.forename: M J
authors.initials: MJ
authors.lastname: Brennan
authors.forename: Z M
authors.initials: ZM
authors.lastname: Li
authors.forename: R D
authors.initials: RD
authors.lastname: Shahin
authors.forename: D L
authors.initials: DL
authors.lastname: Burns
authors.forename: N Y
authors.initials: NY
authors.lastname: Nguyen
authors.forename: T Y
authors.initials: TY
authors.lastname: Liu
authors.forename: M C
authors.initials: MC
authors.lastname: Gray
authors.forename: E L
authors.initials: EL
authors.lastname: Hewlett
authors.forename: C R
authors.initials: CR
authors.lastname: Manclark
mesh_terms: D000067956:Adenylyl Cyclase Inhibitors; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial / Q000302:isolation & purification; D001425:Bacterial Outer Membrane Proteins / Q000276:immunology / Q000302:isolation & purification*; D001886:Bordetella pertussis / Q000276:immunology / Q000302:isolation & purification* / Q000472:pathogenicity; D051379:Mice; D008807:Mice, Inbred BALB C; D008970:Molecular Weight; D010567:Pertussis Vaccine / Q000302:isolation & purification; D014917:Whooping Cough / Q000517:prevention & control
chemical_list: D000067956:Adenylyl Cyclase Inhibitors; D000596:Amino Acids; D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D010567:Pertussis Vaccine