Analysis of a collagen-binding trimeric autotransporter adhesin from Mannheimia haemolytica A1.

A locus that codes for a high-molecular-weight adhesin was previously isolated from Mannheimia haemolytica A1. In this study, we showed that this locus, named ahs, codes for two proteins (AhsA and AhsB) that exhibit characteristics of a trimeric autotransporter adhesin. Sequence analysis of AhsA showed the presence of 21 collagen-binding motifs in the protein. Collagen-binding assays showed that M. haemolytica A1 binds to collagen in a dose-dependent manner. This binding activity is trypsin sensitive and can be inhibited by anti-AhsA antibody. AhsB is the cognate transporter for AhsA. The C-terminal of AhsB showed highly conserved amino acids typical of trimeric autotransporters. Experimental data showed that the C-terminal 120 amino acids of AhsB could indeed form trimeric molecules. Western immunoblots showed the presence of anti-AhsA antibodies in the sera of calves that had been challenged with M. haemolytica A1, suggesting that AhsA is expressed and immunogenic in cattle.

[1]  G. Waksman,et al.  Repetitive Architecture of the Haemophilus influenzae Hia Trimeric Autotransporter , 2008, Journal of molecular biology.

[2]  J. S. St. Geme,et al.  Identification of a Novel Trimeric Autotransporter Adhesin in the Cryptic Genospecies of Haemophilus , 2008, Journal of bacteriology.

[3]  T. Ruiz,et al.  Functional Mapping of an Oligomeric Autotransporter Adhesin of Aggregatibacter actinomycetemcomitans , 2008, Journal of bacteriology.

[4]  P. Speziale,et al.  Functional and structural properties of CbpA, a collagen-binding protein from Arcanobacterium pyogenes. , 2007, Microbiology.

[5]  G. Ulett,et al.  Autotransporter proteins: novel targets at the bacterial cell surface. , 2007, FEMS microbiology letters.

[6]  R. Lo,et al.  The outer membrane protein OmpA of Mannheimia haemolytica A1 is involved in the binding of fibronectin. , 2007, FEMS microbiology letters.

[7]  S. Sathiamoorthy,et al.  Analysis of in vivo expressed genes in Mannheimia haemolytica A1. , 2006, FEMS microbiology letters.

[8]  Milton H. Saier,et al.  Protein-Translocating Trimeric Autotransporters of Gram-Negative Bacteria , 2006, Journal of bacteriology.

[9]  A. Lupas,et al.  Trimeric autotransporter adhesins: variable structure, common function. , 2006, Trends in microbiology.

[10]  J. S. St. Geme,et al.  Trimeric autotransporters: a distinct subfamily of autotransporter proteins. , 2005, Trends in microbiology.

[11]  I. Henderson,et al.  Type V Protein Secretion Pathway: the Autotransporter Story , 2004, Microbiology and Molecular Biology Reviews.

[12]  V. Tenorio,et al.  Adherence of Actinobacillus pleuropneumoniae to swine-lung collagen. , 2004, Microbiology.

[13]  J. Heesemann,et al.  Molecular Analysis of Transport and Oligomerization of the Yersinia enterocolitica Adhesin YadA , 2003, Journal of bacteriology.

[14]  D. Fink,et al.  Immunization with Haemophilus influenzae Hap adhesin protects against nasopharyngeal colonization in experimental mice. , 2002, The Journal of infectious diseases.

[15]  R. Lo,et al.  Construction and Characterization of an Acapsular Mutant of Mannheimia haemolytica A1 , 2002, Infection and Immunity.

[16]  R. Lo.  Genetic analysis of virulence factors of Mannheimia (Pasteurella) haemolytica A1. , 2001, Veterinary microbiology.

[17]  S. Highlander,et al.  Molecular genetic analysis of virulence in Mannheimia (pasteurella) haemolytica. , 2001, Frontiers in bioscience : a journal and virtual library.

[18]  M. Skurnik,et al.  Functional mapping of the Yersinia enterocolitica adhesin YadA. Identification of eight NSVAIG – S motifs in the amino‐terminal half of the protein involved in collagen binding , 2000, Molecular microbiology.

[19]  D. Hodgins,et al.  Vaccination of neonatal colostrum-deprived calves against Pasteurella haemolytica A1. , 2000, Canadian journal of veterinary research = Revue canadienne de recherche veterinaire.

[20]  L. Alphen.  Adhesin-Receptor Interactions by Haemophilus Influenzae and Other Bacteria from the HAP Group , 1995 .

[21]  D. Morck,et al.  Electron microscopic examination of cells of Pasteurella haemolytica-A1 in experimentally infected cattle. , 1988, Canadian journal of veterinary research = Revue canadienne de recherche veterinaire.

[22]  L. Babiuk,et al.  Electron microscopic description of glycocalyx and fimbriae on the surface of Pasteurella haemolytica-A1. , 1987, Canadian journal of veterinary research = Revue canadienne de recherche veterinaire.