Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1996 May;178(10):2876–2882. doi: 10.1128/jb.178.10.2876-2882.1996

Characterization of the binding activities of proteinase-adhesin complexes from Porphyromonas gingivalis.

R N Pike 1, J Potempa 1, W McGraw 1, T H Coetzer 1, J Travis 1
PMCID: PMC178023  PMID: 8631676

Abstract

Adhesins from oral bacteria perform an important function in colonizing target tissues within the dentogingival cavity. In Porphyromonas gingivalis certain of these adhesion proteins exist as a complex with either of two major proteinases referred to as gingipain R (arginine-specific gingipain) and gingipain K (lysine-specific gingipain) (R. N. Pike, W. T. McGraw, J. Potempa, and J. Travis, J. Biol. Chem. 269:406-411, 1994). With specific proteinase inhibitors, it was shown that hemagglutination by either proteinase-adhesin complex could occur independently of proteinase activity. Significantly, low concentrations of fibrinogen, fibronectin, and laminin inhibited hemagglutination, indicating that adherence to these proteins and not the hemagglutination activity was a primary property of the adhesin activity component of complexes. Binding studies with gingipain K and gingipain R suggest that interaction with fibrinogen is a major function of the adhesin domain, with dissociation constants for binding to fibrinogen being 4 and 8.5 nM, respectively. Specific association with fibronectin and laminin was also found. All bound proteins were degraded by the functional proteinase domain, with gingipain R being more active on laminin and fibronectin and gingipain K being more effective in the digestion of fibrinogen. Cumulatively, these data suggest that gingipain R and gingipain K, acting as proteinase-adhesin complexes, progressively attach to, degrade, and detach from target proteins. Since such complexes appear to be present on the surfaces of both vesicles and membranes of P. gingivalis, they may play an important role in the attachment of this bacterium to host cell surfaces.

Full Text

The Full Text of this article is available as a PDF (456.9 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Birkedal-Hansen H., Taylor R. E., Zambon J. J., Barwa P. K., Neiders M. E. Characterization of collagenolytic activity from strains of Bacteroides gingivalis. J Periodontal Res. 1988 Jul;23(4):258–264. doi: 10.1111/j.1600-0765.1988.tb01369.x. [DOI] [PubMed] [Google Scholar]
  2. Chen Z., Potempa J., Polanowski A., Wikstrom M., Travis J. Purification and characterization of a 50-kDa cysteine proteinase (gingipain) from Porphyromonas gingivalis. J Biol Chem. 1992 Sep 15;267(26):18896–18901. [PubMed] [Google Scholar]
  3. Ciborowski P., Nishikata M., Allen R. D., Lantz M. S. Purification and characterization of two forms of a high-molecular-weight cysteine proteinase (porphypain) from Porphyromonas gingivalis. J Bacteriol. 1994 Aug;176(15):4549–4557. doi: 10.1128/jb.176.15.4549-4557.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Coetzer T. H., Pike R. N., Dennison C. Localization of an immunoinhibitory epitope of the cysteine proteinase, cathepsin L. Immunol Invest. 1992 Oct;21(6):495–506. doi: 10.3109/08820139209069386. [DOI] [PubMed] [Google Scholar]
  5. Doolittle R. F., Cassman K. G., Cottrell B. A., Friezner S. J., Takagi T. Amino acid sequence studies on the alpha chain of human fibrinogen. Covalent structure of the alpha-chain portion of fragment D. Biochemistry. 1977 Apr 19;16(8):1710–1715. doi: 10.1021/bi00627a029. [DOI] [PubMed] [Google Scholar]
  6. Glanville R. W., Rauter A., Fietzek P. P. Isolation and characterization of a native placental basement-membrane collagen and its component alpha chains. Eur J Biochem. 1979 Apr 2;95(2):383–389. doi: 10.1111/j.1432-1033.1979.tb12976.x. [DOI] [PubMed] [Google Scholar]
  7. Grenier D., Chao G., McBride B. C. Characterization of sodium dodecyl sulfate-stable Bacteroides gingivalis proteases by polyacrylamide gel electrophoresis. Infect Immun. 1989 Jan;57(1):95–99. doi: 10.1128/iai.57.1.95-99.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Grenier D. Further evidence for a possible role of trypsin-like activity in the adherence of Porphyromonas gingivalis. Can J Microbiol. 1992 Nov;38(11):1189–1192. doi: 10.1139/m92-195. [DOI] [PubMed] [Google Scholar]
  9. Grenier D., McBride B. C. Isolation of a membrane-associated Bacteroides gingivalis glycylprolyl protease. Infect Immun. 1987 Dec;55(12):3131–3136. doi: 10.1128/iai.55.12.3131-3136.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hayashi H., Nagata A., Hinode D., Sato M., Nakamura R. Survey of a receptor protein in human erythrocytes for hemagglutinin of Porphyromonas gingivalis. Oral Microbiol Immunol. 1992 Aug;7(4):204–211. doi: 10.1111/j.1399-302x.1992.tb00026.x. [DOI] [PubMed] [Google Scholar]
  11. Hoover C. I., Ng C. Y., Felton J. R. Correlation of haemagglutination activity with trypsin-like protease activity of Porphyromonas gingivalis. Arch Oral Biol. 1992;37(7):515–520. doi: 10.1016/0003-9969(92)90133-s. [DOI] [PubMed] [Google Scholar]
  12. Imamura T., Pike R. N., Potempa J., Travis J. Pathogenesis of periodontitis: a major arginine-specific cysteine proteinase from Porphyromonas gingivalis induces vascular permeability enhancement through activation of the kallikrein/kinin pathway. J Clin Invest. 1994 Jul;94(1):361–367. doi: 10.1172/JCI117330. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Imamura T., Potempa J., Pike R. N., Moore J. N., Barton M. H., Travis J. Effect of free and vesicle-bound cysteine proteinases of Porphyromonas gingivalis on plasma clot formation: implications for bleeding tendency at periodontitis sites. Infect Immun. 1995 Dec;63(12):4877–4882. doi: 10.1128/iai.63.12.4877-4882.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Inoshita E., Amano A., Hanioka T., Tamagawa H., Shizukuishi S., Tsunemitsu A. Isolation and some properties of exohemagglutinin from the culture medium of Bacteroides gingivalis 381. Infect Immun. 1986 May;52(2):421–427. doi: 10.1128/iai.52.2.421-427.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kay H. M., Birss A. J., Smalley J. W. Haemagglutinating and haemolytic activity of the extracellular vesicles of Bacteroides gingivalis W50. Oral Microbiol Immunol. 1990 Oct;5(5):269–274. doi: 10.1111/j.1399-302x.1990.tb00424.x. [DOI] [PubMed] [Google Scholar]
  16. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  17. Lantz M. S., Allen R. D., Duck L. W., Blume J. L., Switalski L. M., Hook M. Identification of Porphyromonas gingivalis components that mediate its interactions with fibronectin. J Bacteriol. 1991 Jul;173(14):4263–4270. doi: 10.1128/jb.173.14.4263-4270.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lantz M. S., Allen R. D., Vail T. A., Switalski L. M., Hook M. Specific cell components of Bacteroides gingivalis mediate binding and degradation of human fibrinogen. J Bacteriol. 1991 Jan;173(2):495–504. doi: 10.1128/jb.173.2.495-504.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Marsh P. D., McKee A. S., McDermid A. S., Dowsett A. B. Ultrastructure and enzyme activities of a virulent and an avirulent variant of Bacteroides gingivalis W50. FEMS Microbiol Lett. 1989 May;50(1-2):181–185. doi: 10.1016/0378-1097(89)90482-5. [DOI] [PubMed] [Google Scholar]
  20. Mayrand D., Holt S. C. Biology of asaccharolytic black-pigmented Bacteroides species. Microbiol Rev. 1988 Mar;52(1):134–152. doi: 10.1128/mr.52.1.134-152.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Nishikata M., Yoshimura F. Characterization of Porphyromonas (bacteroides) gingivalis hemagglutinin as a protease. Biochem Biophys Res Commun. 1991 Jul 15;178(1):336–342. doi: 10.1016/0006-291x(91)91819-x. [DOI] [PubMed] [Google Scholar]
  22. Nishikata M., Yoshimura F., Nodasaka Y. Possibility of Bacteroides gingivalis hemagglutinin possessing protease activity revealed by inhibition studies. Microbiol Immunol. 1989;33(1):75–80. doi: 10.1111/j.1348-0421.1989.tb01499.x. [DOI] [PubMed] [Google Scholar]
  23. Okuda K., Yamamoto A., Naito Y., Takazoe I., Slots J., Genco R. J. Purification and properties of hemagglutinin from culture supernatant of Bacteroides gingivalis. Infect Immun. 1986 Dec;54(3):659–665. doi: 10.1128/iai.54.3.659-665.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Patti J. M., Allen B. L., McGavin M. J., Hök M. MSCRAMM-mediated adherence of microorganisms to host tissues. Annu Rev Microbiol. 1994;48:585–617. doi: 10.1146/annurev.mi.48.100194.003101. [DOI] [PubMed] [Google Scholar]
  25. Pavloff N., Potempa J., Pike R. N., Prochazka V., Kiefer M. C., Travis J., Barr P. J. Molecular cloning and structural characterization of the Arg-gingipain proteinase of Porphyromonas gingivalis. Biosynthesis as a proteinase-adhesin polyprotein. J Biol Chem. 1995 Jan 20;270(3):1007–1010. doi: 10.1074/jbc.270.3.1007. [DOI] [PubMed] [Google Scholar]
  26. Pike R., McGraw W., Potempa J., Travis J. Lysine- and arginine-specific proteinases from Porphyromonas gingivalis. Isolation, characterization, and evidence for the existence of complexes with hemagglutinins. J Biol Chem. 1994 Jan 7;269(1):406–411. [PubMed] [Google Scholar]
  27. Potempa J., Pike R., Travis J. The multiple forms of trypsin-like activity present in various strains of Porphyromonas gingivalis are due to the presence of either Arg-gingipain or Lys-gingipain. Infect Immun. 1995 Apr;63(4):1176–1182. doi: 10.1128/iai.63.4.1176-1182.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Progulske-Fox A., Rao V., Han N., Lepine G., Witlock J., Lantz M. Molecular characterization of hemagglutinin genes of periodontopathic bacteria. J Periodontal Res. 1993 Nov;28(6 Pt 2):473–474. doi: 10.1111/j.1600-0765.1993.tb02106.x. [DOI] [PubMed] [Google Scholar]
  29. Schägger H., von Jagow G. Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal Biochem. 1987 Nov 1;166(2):368–379. doi: 10.1016/0003-2697(87)90587-2. [DOI] [PubMed] [Google Scholar]
  30. Scott C. F., Whitaker E. J., Hammond B. F., Colman R. W. Purification and characterization of a potent 70-kDa thiol lysyl-proteinase (Lys-gingivain) from Porphyromonas gingivalis that cleaves kininogens and fibrinogen. J Biol Chem. 1993 Apr 15;268(11):7935–7942. [PubMed] [Google Scholar]
  31. Slots J., Listgarten M. A. Bacteroides gingivalis, Bacteroides intermedius and Actinobacillus actinomycetemcomitans in human periodontal diseases. J Clin Periodontol. 1988 Feb;15(2):85–93. doi: 10.1111/j.1600-051x.1988.tb00999.x. [DOI] [PubMed] [Google Scholar]
  32. Smalley J. W., Birss A. J., Kay H. M., McKee A. S., Marsh P. D. The distribution of trypsin-like enzyme activity in cultures of a virulent and an avirulent strain of Bacteroides gingivalis W50. Oral Microbiol Immunol. 1989 Sep;4(3):178–181. doi: 10.1111/j.1399-302x.1989.tb00249.x. [DOI] [PubMed] [Google Scholar]
  33. Travis J., Pannell R. Selective removal of albumin from plasma by affinity chromatography. Clin Chim Acta. 1973 Nov 23;49(1):49–52. doi: 10.1016/0009-8981(73)90341-0. [DOI] [PubMed] [Google Scholar]
  34. van Winkelhoff A. J., van Steenbergen T. J., de Graaff J. The role of black-pigmented Bacteroides in human oral infections. J Clin Periodontol. 1988 Mar;15(3):145–155. doi: 10.1111/j.1600-051x.1988.tb01561.x. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES