Outer membrane proteins of bovine Pasteurella multocida serogroup A isolates

• Published in: Veterinary microbiology Vol. 54; no. 2; pp. 167 - 183
• Main Authors: Dabo, S.M., Confer, A.W., Murphy, G.L.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 01.02.1997; Elsevier Science
• Subjects: Amino Acid Sequence; Animals; ANTIBODIES; ANTICORPS; ANTICUERPOS; ANTIGENE; ANTIGENOS; ANTIGENS; Bacterial Outer Membrane Proteins - analysis; Bacterial Outer Membrane Proteins - chemistry; Bacteriology; Biological and medical sciences; Blotting, Western; BOVIN; CATTLE; Cattle Diseases; Centrifugation, Density Gradient; CERDO; Electrophoresis, Polyacrylamide Gel; Fundamental and applied biological sciences. Psychology; GANADO BOVINO; Gram-Negative Bacteria; Microbiology; Molecular Sequence Data; N-terminal amino acid; Outer membrane protein; Pasteurella Infections - diagnosis; Pasteurella Infections - veterinary; PASTEURELLA MULTOCIDA; Pasteurella multocida - classification; Pasteurella multocida - isolation & purification; Pathogenicity, virulence, toxins, bacteriocins, pyrogens, host-bacteria relations, miscellaneous strains; PORCIN; Protease; PROTEINAS; PROTEINE; PROTEINS; Sequence Homology, Amino Acid; Serotyping; Sucrose density gradient; Surface exposure; SWINE; Surface exposure; Protease; Sucrose density gradient; Outer membrane protein; N-terminal amino acid; Pasteurella multocida; Membrane protein; Bovine; Host agent relation; External membrane; Thermal stability; Pasteurellaceae; Characterization; Vertebrata; Mammalia; Antigenicity; Immunogenicity; Animal; Bacteria; Serum; Artiodactyla; Veterinary; Ungulata
• ISSN: 0378-1135; 1873-2542
• DOI: 10.1016/S0378-1135(96)01274-6

The outer membrane proteins (OMPs) of P. multocida serotypes A3 (7 isolates), A4 (2 isolates), A3,4 and A2 (one isolate each) obtained from pneumonic cattle (10 isolates) and from one pig isolate were investigated to identify potential immunogens. SDS-PAGE of P. multocida OM isolated by SDG centrifugation of spheroplasts revealed eight major OMPs. Outer membranes isolated by sarcosyl extraction or SDG had similar protein composition on Coomassie blue-stained SDS-PA gel and on immunoblots. Two major OMPs ( M rs of 35 and 46 kDa at 100°C) demonstrated heat modifiability with apparent M rs of 30 and 34 kDa at 37°C, respectively. The N-terminal aa sequences of these heat modifiable proteins revealed homology with E. coli OmpA and Hib P1 proteins, respectively. Protease treatment of whole cells followed by western immunoblots using bovine convalescent sera identified several immunogenic, surface-exposed and conserved OMPs among the eleven P. multocida isolates examined. The whole organism SDS-PAGE profiles of the eleven P. multocida isolates differed such that six patterns were seen. These patterns could potentially be used as a typing system for P. multocida bovine isolates based on the molecular weights of whole cell proteins. The above observations have potentially important implications relative to the immunity to infection.