title: The 32 kDa major outer-membrane protein of Pasteurella multocida capsular serotype D.
abstract: The major outer-membrane protein (MOMP) of Pasteurella multocida serotype D strain P210, with an apparent molecular mass of 32 kDa, was purified and characterized. The purification method involved selective extraction of MOMP with N-lauroylsarcosine and SDS, followed by immunoaffinity chromatography using a murine monoclonal antibody (mAb). The N-terminal sequence and amino acid composition of the MOMP showed considerable similarity to other Gram-negative bacterial porins, notably to the 37 kDa MOMP (porin H) of P. multocida. Immunoelectron microscopy and colony blotting assays were used to demonstrate the surface localization of the 32 kDa MOMP on bacterial cells. The colony blotting assay provided a simple, sensitive and rapid screening method for visualizing accessibility of the antibody on the cells. In a Western blot assay, murine polyclonal hyperimmune serum against the purified 32 kDa MOMP recognized both serotype B and D strains bearing either a 32 kDa or a 37 kDa MOMP, whereas the mAb recognized only serotype D strains bearing a 32 kDa but not a 37 kDa MOMP. The present data indicate that the 32 kDa MOMPs of P. multocida are antigenically heterogeneous and possess both specific and cross-reacting epitopes. Detection of type-specific epitopes on the 32 kDa MOMP using an mAb may have potential implications regarding the feasibility of developing a serotyping system for P. multocida.
authors.affiliation: Département de Pathologie et Microbiologie, Faculté de Médécine Vétérinaire, Université de Montréal, CP 5000, Saint-Hyacinthe, Québec, Canada J2S 7C6.
authors.forename: M
authors.initials: M
authors.lastname: Vasfi Marandi
authors.affiliation: Département de Pathologie et Microbiologie, Faculté de Médécine Vétérinaire, Université de Montréal, CP 5000, Saint-Hyacinthe, Québec, Canada J2S 7C6.
authors.forename: J D
authors.initials: JD
authors.lastname: Dubreuil
authors.affiliation: Département de Pathologie et Microbiologie, Faculté de Médécine Vétérinaire, Université de Montréal, CP 5000, Saint-Hyacinthe, Québec, Canada J2S 7C6.
authors.forename: K R
authors.initials: KR
authors.lastname: Mittal
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D016667:Bacterial Capsules / Q000276:immunology; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification*; D015153:Blotting, Western; D002451:Cell Compartmentation; D002846:Chromatography, Affinity; D016253:Microscopy, Immunoelectron; D008969:Molecular Sequence Data; D016979:Pasteurella multocida / Q000737:chemistry* / Q000145:classification / Q000276:immunology; D018272:Porins; D017421:Sequence Analysis; D017386:Sequence Homology, Amino Acid; D012703:Serotyping; D013045:Species Specificity
chemical_list: D000596:Amino Acids; D001425:Bacterial Outer Membrane Proteins; D018272:Porins; C067128:ompH protein, bacteria