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Infection and Immunity logoLink to Infection and Immunity
. 1996 Jul;64(7):2532–2539. doi: 10.1128/iai.64.7.2532-2539.1996

Characterization of an adherence and antigenic determinant of the ArgI protease of Porphyromonas gingivalis which is present on multiple gene products.

M A Curtis 1, J Aduse-Opoku 1, J M Slaney 1, M Rangarajan 1, V Booth 1, J Cridland 1, P Shepherd 1
PMCID: PMC174107  PMID: 8698476

Abstract

This study was performed to characterize the antigen(s) recognized by a panel of monoclonal antibodies (MAbs) produced to be specific for Porphyromonas gingivalis whole cells which we had previously shown to bind to epitopes recognized by sera from periodontitis patients. Preliminary data had suggested that the arginine-specific proteases of P. gingivalis (ArgI, ArgIA, and ArgIB) contained the antigenic determinants of four of these antibodies (MAbs 1A1, 2B/H9, 7D5, and 3B1). The location of the binding sites was examined with purified P. gingivalis enzymes and recombinant regions of the ArgI polyprotein expressed by subclones of the prpR1 gene in Escherichia coli XL-1 Blue cells. All four antibodies were reactive with protein determinants within the beta subunit, a hemagglutinin and/or adhesin component, of the ArgI dimer. MAb 1A1 strongly inhibited the agglutination of human erythrocytes by P. gingivalis W50 culture supernatant, suggesting that the binding site for this antibody contains residues which are critical for the interaction with the erythrocyte surface. The determinant for MAb 1A1 was examined further by construction of a set of truncated forms of the beta component expressed as fusion proteins with glutathione S-transferase at the N terminus. Analysis of these constructs mapped the binding site for MAb 1A1 to PrpRI residues G-907 to T-931, GVSPKVCKDV TVEGSNEFAP VQNLT. Western blot (immunoblot) analysis of P. gingivalis whole-cell proteins demonstrated that MAb 1A1 reacts with several proteins in the Mr range of 20,000 to 120,000. Furthermore, an oligonucleotide probe corresponding to the coding sequence for the region of the ArgI beta component containing the MAb 1A1 binding site hybridized to multiple bands on genomic digests of P. gingivalis DNA. These data indicate that the MAb 1A1 epitope may be a component of a binding domain common to multiple gene products of this organism and may thus represent a functionally important target of the host's specific immune response to P. gingivalis in periodontal disease.

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Selected References

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  1. Aduse-Opoku J., Muir J., Slaney J. M., Rangarajan M., Curtis M. A. Characterization, genetic analysis, and expression of a protease antigen (PrpRI) of Porphyromonas gingivalis W50. Infect Immun. 1995 Dec;63(12):4744–4754. doi: 10.1128/iai.63.12.4744-4754.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Boyd J., McBride B. C. Fractionation of hemagglutinating and bacterial binding adhesins of Bacteroides gingivalis. Infect Immun. 1984 Aug;45(2):403–409. doi: 10.1128/iai.45.2.403-409.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chandad F., Mouton C. Antigenic, structural, and functional relationships between fimbriae and the hemagglutinating adhesin HA-Ag2 of Porphyromonas gingivalis. Infect Immun. 1995 Dec;63(12):4755–4763. doi: 10.1128/iai.63.12.4755-4763.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cridland J. C., Booth V., Ashley F. P., Curtis M. A., Wilson R. F., Shepherd P. Preliminary characterisation of antigens recognised by monoclonal antibodies raised to Porphyromonas gingivalis and by sera from patients with periodontitis. J Periodontal Res. 1994 Sep;29(5):339–347. doi: 10.1111/j.1600-0765.1994.tb01232.x. [DOI] [PubMed] [Google Scholar]
  5. Curtis M. A., Ramakrishnan M., Slaney J. M. Characterization of the trypsin-like enzymes of Porphyromonas gingivalis W83 using a radiolabelled active-site-directed inhibitor. J Gen Microbiol. 1993 May;139(5):949–955. doi: 10.1099/00221287-139-5-949. [DOI] [PubMed] [Google Scholar]
  6. Curtis M. A., Slaney J. M., Carman R. J., Johnson N. W. Identification of the major surface protein antigens of Porphyromonas gingivalis using IgG antibody reactivity of periodontal case-control serum. Oral Microbiol Immunol. 1991 Dec;6(6):321–326. doi: 10.1111/j.1399-302x.1991.tb00502.x. [DOI] [PubMed] [Google Scholar]
  7. Cutler C. W., Arnold R. R., Schenkein H. A. Inhibition of C3 and IgG proteolysis enhances phagocytosis of Porphyromonas gingivalis. J Immunol. 1993 Dec 15;151(12):7016–7029. [PubMed] [Google Scholar]
  8. Ebersole J. L., Brunsvold M., Steffensen B., Wood R., Holt S. C. Effects of immunization with Porphyromonas gingivalis and Prevotella intermedia on progression of ligature-induced periodontitis in the nonhuman primate Macaca fascicularis. Infect Immun. 1991 Oct;59(10):3351–3359. doi: 10.1128/iai.59.10.3351-3359.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ebersole J. L., Taubman M. A., Smith D. J., Frey D. E. Human immune responses to oral microorganisms: patterns of systemic antibody levels to Bacteroides species. Infect Immun. 1986 Feb;51(2):507–513. doi: 10.1128/iai.51.2.507-513.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Ellen R. P., Song M., Buivids I. A. Inhibition of Actinomyces viscosus--Porphyromonas gingivalis coadhesion by trypsin and other proteins. Oral Microbiol Immunol. 1992 Aug;7(4):198–203. doi: 10.1111/j.1399-302x.1992.tb00025.x. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. 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]
  13. Isogai H., Isogai E., Yoshimura F., Suzuki T., Kagota W., Takano K. Specific inhibition of adherence of an oral strain of Bacteroides gingivalis 381 to epithelial cells by monoclonal antibodies against the bacterial fimbriae. Arch Oral Biol. 1988;33(7):479–485. doi: 10.1016/0003-9969(88)90028-3. [DOI] [PubMed] [Google Scholar]
  14. Johnson V., Johnson B. D., Sims T. J., Whitney C. W., Moncla B. J., Engel L. D., Page R. C. Effects of treatment on antibody titer to Porphyromonas gingivalis in gingival crevicular fluid of patients with rapidly progressive periodontitis. J Periodontol. 1993 Jun;64(6):559–565. doi: 10.1902/jop.1993.64.6.559. [DOI] [PubMed] [Google Scholar]
  15. Kirszbaum L., Sotiropoulos C., Jackson C., Cleal S., Slakeski N., Reynolds E. C. Complete nucleotide sequence of a gene prtR of Porphyromonas gingivalis W50 encoding a 132 kDa protein that contains an arginine-specific thiol endopeptidase domain and a haemagglutinin domain. Biochem Biophys Res Commun. 1995 Feb 6;207(1):424–431. doi: 10.1006/bbrc.1995.1205. [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. Lee J. Y., Sojar H. T., Bedi G. S., Genco R. J. Synthetic peptides analogous to the fimbrillin sequence inhibit adherence of Porphyromonas gingivalis. Infect Immun. 1992 Apr;60(4):1662–1670. doi: 10.1128/iai.60.4.1662-1670.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Millar D. J., Scott E. E., Slaney J. M., U S., Benjamin P., Curtis M. A. Production and characterisation of monoclonal antibodies to the principle sonicate antigens of Porphyromonas gingivalis w50. FEMS Immunol Med Microbiol. 1993 Oct;7(3):211–222. doi: 10.1111/j.1574-695X.1993.tb00401.x. [DOI] [PubMed] [Google Scholar]
  21. Mouton C., Bouchard D., Deslauriers M., Lamonde L. Immunochemical identification and preliminary characterization of a nonfimbrial hemagglutinating adhesin of Bacteroides gingivalis. Infect Immun. 1989 Feb;57(2):566–573. doi: 10.1128/iai.57.2.566-573.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Naito Y., Gibbons R. J. Attachment of Bacteroides gingivalis to collagenous substrata. J Dent Res. 1988 Aug;67(8):1075–1080. doi: 10.1177/00220345880670080301. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. Okamoto K., Misumi Y., Kadowaki T., Yoneda M., Yamamoto K., Ikehara Y. Structural characterization of argingipain, a novel arginine-specific cysteine proteinase as a major periodontal pathogenic factor from Porphyromonas gingivalis. Arch Biochem Biophys. 1995 Feb 1;316(2):917–925. doi: 10.1006/abbi.1995.1123. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. 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]
  27. 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]
  28. Progulske-Fox A., Tumwasorn S., Holt S. C. The expression and function of a Bacteroides gingivalis hemagglutinin gene in Escherichia coli. Oral Microbiol Immunol. 1989 Sep;4(3):121–131. doi: 10.1111/j.1399-302x.1989.tb00238.x. [DOI] [PubMed] [Google Scholar]
  29. Progulske-Fox A., Tumwasorn S., Lépine G., Whitlock J., Savett D., Ferretti J. J., Banas J. A. The cloning, expression and sequence analysis of a second Porphyromonas gingivalis gene that codes for a protein involved in hemagglutination. Oral Microbiol Immunol. 1995 Oct;10(5):311–318. doi: 10.1111/j.1399-302x.1995.tb00160.x. [DOI] [PubMed] [Google Scholar]
  30. Schenkein H. A., Fletcher H. M., Bodnar M., Macrina F. L. Increased opsonization of a prtH-defective mutant of Porphyromonas gingivalis W83 is caused by reduced degradation of complement-derived opsonins. J Immunol. 1995 May 15;154(10):5331–5337. [PubMed] [Google Scholar]
  31. 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]
  32. Slots J., Bragd L., Wikström M., Dahlén G. The occurrence of Actinobacillus actinomycetemcomitans, Bacteroides gingivalis and Bacteroides intermedius in destructive periodontal disease in adults. J Clin Periodontol. 1986 Jul;13(6):570–577. doi: 10.1111/j.1600-051x.1986.tb00849.x. [DOI] [PubMed] [Google Scholar]
  33. Suzuki Y., Yoshimura F., Takahashi K., Tani H., Suzuki T. Detection of fimbriae and fimbrial antigens on the oral anaerobe Bacteroides gingivalis by negative staining and serological methods. J Gen Microbiol. 1988 Oct;134(10):2713–2720. doi: 10.1099/00221287-134-10-2713. [DOI] [PubMed] [Google Scholar]
  34. Suzuki Y., Yoshimura F., Tani H., Suzuki T. Fimbriae from the oral anaerobe Bacteroides gingivalis: a screening of clinical isolates from various places. Adv Dent Res. 1988 Nov;2(2):301–303. doi: 10.1177/08959374880020021601. [DOI] [PubMed] [Google Scholar]
  35. Watanabe H., Marsh P. D., Ivanyi L. Detection of immunodominant antigens of periodontopathic bacteria in human periodontal disease. Oral Microbiol Immunol. 1989 Sep;4(3):159–164. doi: 10.1111/j.1399-302x.1989.tb00244.x. [DOI] [PubMed] [Google Scholar]
  36. Westerlund B., Korhonen T. K. Bacterial proteins binding to the mammalian extracellular matrix. Mol Microbiol. 1993 Aug;9(4):687–694. doi: 10.1111/j.1365-2958.1993.tb01729.x. [DOI] [PubMed] [Google Scholar]
  37. Wilton J. M., Johnson N. W., Curtis M. A., Gillett I. R., Carman R. J., Bampton J. L., Griffiths G. S., Sterne J. A. Specific antibody responses to subgingival plaque bacteria as aids to the diagnosis and prognosis of destructive periodontitis. J Clin Periodontol. 1991 Jan;18(1):1–15. doi: 10.1111/j.1600-051x.1991.tb01112.x. [DOI] [PubMed] [Google Scholar]
  38. Wingrove J. A., DiScipio R. G., Chen Z., Potempa J., Travis J., Hugli T. E. Activation of complement components C3 and C5 by a cysteine proteinase (gingipain-1) from Porphyromonas (Bacteroides) gingivalis. J Biol Chem. 1992 Sep 15;267(26):18902–18907. [PubMed] [Google Scholar]
  39. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]

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