1887

Abstract

, an opportunistic pathogen of economically important food animals, is the causative agent of liver abscesses in feedlot cattle, osteomyelitis in turkeys, and pneumonia and arthritis in pigs. Previous studies identified the first adhesin, CbpA, a protein located on the bacterial surface which has the ability to bind collagen and promotes adhesion to the host cells. The protein has an N-terminal ligand-binding region (region A) and a C-terminal repetitive domain (region B). In this study we found that CbpA bound to almost all the collagen types tested but not to other proteins, and it displayed a propensity to interact with several collagenous peptides derived by CNBr cleavage of type I and II collagens. The values of CbpA for type I and II collagens and collagen peptides determined by solid-phase binding assay and intrinsic tryptophan fluorescence were in the range of 1–15 nM. It was also found that CbpA and its A region bound fibronectin, and that collagen and fibronectin interacted with distinct subsites. Anti-CbpA antibodies were effective at inhibiting both binding of isolated CbpA and bacterial adhesion to immobilized collagen, suggesting that CbpA is a functional collagen-binding adhesin. Analysis of the immunological cross-reactivity of CbpA with antibodies against other bacterial collagen-binding proteins indicated that CbpA is immunologically related to ACE from but not to CNA from S or Acm from . Far-UV and near-UV circular dichroism spectra showed that full-length CbpA and its region A are mainly composed of -sheet with only a minor -helical component and that both the proteins have a well-defined tertiary structure.

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2007-10-01
2024-12-14
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References

  1. Antikainen J., Anton L., Sillanpää J., Korhonen T. K. 2002; Domains in the S-layer protein CbsA of Lactobacillus crispatus involved in adherence to collagens, laminin and lipoteichoic acids and in self-assembly. Mol Microbiol 46:381–394
    [Google Scholar]
  2. Billington S. J., Jost B. H., Cuevas W. A., Bright K. R., Songer J. G. 1997; The Arcanobacterium ( Actinomyces ) pyogenes hemolysin, pyolysin, is a novel member of the thiol-activated cytolysin family. J Bacteriol 179:6100–6106
    [Google Scholar]
  3. Deivanayagam C. C., Rich R. R., Carson M., Owens R. T., Danthuluri S., Bice T., Höök M., Narayana S. V. 2000; Novel fold and assembly of the repetitive B region of the Staphylococcus aureus collagen-binding surface protein. Structure 8:67–78
    [Google Scholar]
  4. Dinkla K., Rohde M., Jansen W. T., Kaplan E. L., Chhatwal G. S., Talay S. R. 2003; Rheumatic-fever associated Streptococcus pyogenes isolates aggregate collagen. J Clin Invest 111:1905–1912
    [Google Scholar]
  5. Eftink M. R., Ghiron C. A. 1981; Fluorescence quenching studies with proteins. Anal Biochem 114:199–227
    [Google Scholar]
  6. Esmay P. A., Billington S. J., Link M. A., Songer J. G., Jost B. H. 2003; The Arcanobacterium pyogenes collagen binding protein, CbpA, promotes adhesion to host cells. Infect Immun 71:4368–4374
    [Google Scholar]
  7. Foster T. J., Höök M. 1998; Surface protein adhesins of Staphylococcus aureus . Trends Microbiol 6:484–488
    [Google Scholar]
  8. Hubble T. S., Hatton J. F., Nallapareddy S. R., Murray B. E., Gillespie M. J. 2003; Influence of Enterococcus faecalis proteases and the collagen-binding protein, Ace, on adhesion to dentin. Oral Microbiol Immunol 18:121–126
    [Google Scholar]
  9. Huszar G., Maiocco J., Naftolin F. 1980; Monitoring of collagen and collagen fragments in chromatography of protein mixtures. Anal Biochem 105:424–429
    [Google Scholar]
  10. Jost B. H., Billington S. J. 2005; Arcanobacterium pyogenes : molecular pathogenesis of an animal opportunist. Antonie Van Leeuwenhoek 88:87–102
    [Google Scholar]
  11. Jost B. H., Songer J. G., Billington S. J. 2002; Identification of a second Arcanobacterium pyogenes neuraminidase and involvement of neuraminidase activity in host cell adhesion. Infect Immun 70:1106–1112
    [Google Scholar]
  12. Köhler G., Milstein C. 1975; Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 256:495–497
    [Google Scholar]
  13. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
    [Google Scholar]
  14. Nallapareddy S. R., Qin X., Weinstock G. M., Höök M., Murray B. E. 2000; Enterococcus faecalis adhesin, Ace, mediates attachment to extracellular matrix proteins collagen type IV and laminin as well as collagen type I. Infect Immun 68:5218–5224
    [Google Scholar]
  15. Nallapareddy S. R., Weinstock G. M., Murray B. E. 2003; Clinical isolates of Enterococcus faecium exhibit strain-specific collagen binding mediated by Acm, a new member of the MSCRAMM family. Mol Microbiol 47:1733–1747
    [Google Scholar]
  16. Nilsson I. M., Patti J. M., Bremell T., Höök M., Tarkowski A. 1998; Vaccination with a recombinant fragment of collagen adhesin provides protection against Staphylococcus aureus -mediated septic death. J Clin Invest 101:2640–2649
    [Google Scholar]
  17. Nitsche D. P., Johansson H. M., Frick I.-M., Mörgelin M. 2006; Streptococcal protein FOG, a novel matrix adhesin interacting with collagen I in vivo . J Biol Chem 281:1670–1679
    [Google Scholar]
  18. Nummelin H., Merckel M. C., Leo J. C., Lankinen H., Skurnik M., Goldman A. 2004; The Yersinia adhesin Yad collagen-binding domain structure is a novel left-handed parallel β -roll. EMBO J 23:701–711
    [Google Scholar]
  19. Patti J. M., Jonsson H., Guss B., Switalski L. M., Wiberg K., Lindberg M., Höök M. 1992; Molecular characterization and expression of a gene encoding a Staphylococcus aureus collagen adhesin. J Biol Chem 267:4766–4772
    [Google Scholar]
  20. Patti J. M., Boles J. O., Höök M. 1993; Identification and biochemical characterization of the ligand binding domain of the collagen adhesin from Staphylococcus aureus . Biochemistry 32:11428–11435
    [Google Scholar]
  21. Reese C. A., Mayne R. 1981; Minor collagens of chicken hyaline cartilage. Biochemistry 20:5443–5448
    [Google Scholar]
  22. Rich R. L., Demeler B., Ashby K., Deivanayagam C. C., Petrich J. W., Patti J. M., Narayana S. V., Höök M. 1998; Domain structure of the Staphylococcus aureus collagen adhesin. Biochemistry 37:15423–15433
    [Google Scholar]
  23. Rich R. L., Kreikemeyer B., Owens R. T., LaBrenz S., Narayana S. V., Weinstock G. M., Murray B. E., Höök M. 1999; Ace is a collagen-binding MSCRAMM from Enterococcus faecalis . J Biol Chem 274:26939–26945
    [Google Scholar]
  24. Roche F. M., Downer R., Keane F., Speziale P., Park P. W., Foster T. J. 2004; The N-terminal A domain of fibronectin-binding proteins A and B promotes adhesion of Staphylococcus aureus to elastin. J Biol Chem 279:38433–38440
    [Google Scholar]
  25. Rossi A., Vitellaro-Zuccarello L., Zanaboni G., Monzani E., Dyne K. M., Cetta G., Tenni R. 1996; Type I collagen CNBr peptides: species and behavior in solution. Biochemistry 35:6048–6057
    [Google Scholar]
  26. Schaufuss P., Sting R., Lämmler C. 1989; Isolation and characterization of an extracellular protease of Actinomyces pyogenes . Zentralbl Bakteriol 271:452–459
    [Google Scholar]
  27. Shimoji Y., Ogawa Y., Osaki M., Kabeya H., Maruyama S., Mikami T., Sekizaki T. 2003; Adhesive surface proteins of Erysipelothrix rhusiopathiae bind to polystyrene, fibronectin, and type I and IV collagens. J Bacteriol 185:2739–2748
    [Google Scholar]
  28. Visai L., Xu Y., Casolini F., Rindi S., Höök M., Speziale P. 2000; Monoclonal antibodies to CNA, a collagen-binding microbial surface component recognizing adhesive matrix molecules, detach Staphylococcus aureus from a collagen substrate. J Biol Chem 275:39837–39845
    [Google Scholar]
  29. Vuento M., Vaheri A. 1979; Purification of fibronectin from human plasma by affinity chromatography under non-denaturing conditions. Biochem J 183:331–337
    [Google Scholar]
  30. Walsh E. J., O'Brien L. M., Liang X., Höök M., Foster T. J. 2004; Clumping factor B, a fibrinogen-binding MSCRAMM (microbial surface components recognizing adhesive matrix molecules) adhesin of Staphylococcus aureus also binds to the tail region of type I cytokeratin 10. J Biol Chem 279:50691–50699
    [Google Scholar]
  31. Xu Y., Rivas J. M., Brown E. L., Liang X., Höök M. 2004a; Virulence potential of the staphylococcal adhesin CNA in experimental arthritis is determined by its affinity for collagen. J Infect Dis 189:2323–2333
    [Google Scholar]
  32. Xu Y., Liang X., Chen Y., Koehler T. M., Höök M. 2004b; Identification and biochemical characterization of two novel collagen-binding MSCRAMMs of Bacillus anthracis . J Biol Chem 279:51760–51768
    [Google Scholar]
  33. Zong Y., Xu Y., Liang X., Keene D. R., Höök A., Gurusiddappa S., Höök M., Narayana S. V. 2005; A ‘Collagen Hug’ model for Staphylococcus aureus CNA binding to collagen. EMBO J 24:4224–4236
    [Google Scholar]
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