Characterization of a large transferrin-binding protein from Actinobacillus pleuropneumoniae serotype 7.
The binding of transferrin at the surface of Actinobacillus pleuropneumoniae (A. pp.) is mediated by two proteins of approximately 60 and 100 kDa. The 60 kDa protein has been shown to be highly divergent among different serotypes and to induce a serotype-specific protective immune response. In this study we have characterized the 100 kDa transferrin-binding protein of A. pp. serotype 7 and designated it as TfbB. The tfbB gene was found to be located immediately downstream of the tfbA gene. It was cloned and sequenced, and antibodies raised against the isolated recombinant protein detected, with a constant intensity, a 100 kDa protein in A. pp. serotypes 2, 4, 6, 7, 8, 9, 10 and 11, and a polypeptide of approximately 103 kDa in serotypes 1, 3, 5A and 12. In addition, comparative analysis of the deduced amino acid sequence showed more than 40% identity with the large transferrin-binding proteins of Neisseria meningitidis and Haemophilus influenzae. The TfbB protein was expressed in E. coli outer membranes in a conformation eliciting porcine transferrin-specific binding activity. Sera of pigs immunized with these TfbB-containing E. coli membranes recognized functional membrane-associated TfbB protein whereas no such reaction was observed upon immunization with isolated recombinant TfbB protein. A preliminary animal experiment showed that TfbB-containing outer membrane preparations from recombinant E. coli can reduce significantly the mortality of an A.pp. infection with the homologous strain.
Wilke
M
M
Institut für Mikrobiologie und Tierseuchen, Tierärztliche Hochschule Hannover, Germany.
Franz
B
B
Gerlach
G F
GF
D016977:Actinobacillus pleuropneumoniae / Q000737:chemistry* / Q000145:classification / Q000235:genetics; D000595:Amino Acid Sequence; D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000032:analysis / Q000235:genetics / Q000276:immunology; D001426:Bacterial Proteins / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D002352:Carrier Proteins / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D006193:Haemophilus influenzae / Q000737:chemistry / Q000235:genetics; D007114:Immunization / Q000662:veterinary; D033862:Iron-Binding Proteins; D008297:Male; D008969:Molecular Sequence Data; D009345:Neisseria meningitidis / Q000737:chemistry / Q000235:genetics; D011817:Rabbits; D011994:Recombinant Proteins / Q000032:analysis / Q000235:genetics / Q000276:immunology; D017386:Sequence Homology, Amino Acid; D012703:Serotyping / Q000662:veterinary; D013045:Species Specificity; D013552:Swine; D033863:Transferrin-Binding Proteins
D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D002352:Carrier Proteins; D033862:Iron-Binding Proteins; D011994:Recombinant Proteins; D033863:Transferrin-Binding Proteins