• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of iaiPermissionsJournals.ASM.orgJournalIAI ArticleJournal InfoAuthorsReviewers
Infect Immun. Mar 1996; 64(3): 952–958.
PMCID: PMC173862

Selection and phenotypic characterization of nonhemagglutinating mutants of Porphyromonas gingivalis.


To further investigate the relationship between fimbriae and the hemagglutinating adhesin HA-Ag2 of Porphyromonas gingivalis, three spontaneous mutants of the type strain ATCC 33277 were selected by a hemadsorption procedure. They were characterized for hemagglutination, trypsin-like and lectin-binding activities, and hydrophobicity and for the presence of fimbriae. The presence of the 42-kDa (the fimbrilin subunit) and the 43- and 49-kDa (the HA-Ag2 components) polypeptides was investigated by immunoblotting using polyclonal and monoclonal antibodies directed to fimbriae and to the hemagglutinating adhesin HA-Ag2. Cells from two of the three mutants (M1 and M2) exhibited no or little hemagglutination activity and very low trypsin-like activity and did not show the 43- and 49-kDa polypeptides. Abnormal fimbriation in M1 was deduced from the following observations of cells grown for 18 h: absence of the 42-kDa polypeptide and of a 14-kDa polypeptide and no fimbriae visible on electron micrographs. While the cells of mutant M2, irrespective of the age of the culture, were found to lack the 43- and 49-kDa polypeptides and hemagglutination activity, the supernatants of cultures grown for 72 h had high hemagglutination and trypsin-like activities and revealed the presence of the 42-, 43-, and 49-kDa polypeptides. This suggests that M2 may be missing some molecules which anchor the components to the cell surface. Mutant M3 showed levels of activities similar to those of the parental strain but lacked the 43-kDa polypeptide. Other pleiotropic effects observed for the mutants included loss of dark pigmentation and lower hydrophobicity. The data from this study fuel an emerging consensus whereby fimbriation, hemagglutination, and proteolytic activities, as well as other functions in P. gingivalis, are intricate.

Full Text

The Full Text of this article is available as a PDF (808K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Ansorge W. Fast and sensitive detection of protein and DNA bands by treatment with potassium permanganate. J Biochem Biophys Methods. 1985 May;11(1):13–20. [PubMed]
  • Boyd J, McBride BC. Fractionation of hemagglutinating and bacterial binding adhesins of Bacteroides gingivalis. Infect Immun. 1984 Aug;45(2):403–409. [PMC free article] [PubMed]
  • Chandad F, Mouton C. Molecular size variation of the hemagglutinating adhesin HA-Ag2, a common antigen of Bacteroides gingivalis. Can J Microbiol. 1990 Oct;36(10):690–696. [PubMed]
  • 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. [PMC free article] [PubMed]
  • Deslauriers M, Mouton C. Targetting the production of monoclonal antibodies to the hemagglutinating adhesin of Bacteroides (Porphyromonas) gingivalis by injection of an immunoprecipitate from crossed immunoelectrophoresis. J Biol Buccale. 1991 Jun;19(2):155–160. [PubMed]
  • Deslauriers M, Mouton C. Epitope mapping of hemagglutinating adhesion HA-Ag2 of Bacteroides (Porphyromonas) gingivalis. Infect Immun. 1992 Jul;60(7):2791–2799. [PMC free article] [PubMed]
  • Goulbourne PA, Ellen RP. Evidence that Porphyromonas (Bacteroides) gingivalis fimbriae function in adhesion to Actinomyces viscosus. J Bacteriol. 1991 Sep;173(17):5266–5274. [PMC free article] [PubMed]
  • 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. [PubMed]
  • Grenier D, Labbé S, Mouton C, Meyrand D. Hydrolytic enzymes and lectin-binding activity of black-pigmented anaerobic rods. Microbiology. 1994 Apr;140(Pt 4):873–878. [PubMed]
  • Hoover CI, Ng CY, Felton JR. Correlation of haemagglutination activity with trypsin-like protease activity of Porphyromonas gingivalis. Arch Oral Biol. 1992;37(7):515–520. [PubMed]
  • Hoover CI, Yoshimura F. Transposon-induced pigment-deficient mutants of Porphyromonas gingivalis. FEMS Microbiol Lett. 1994 Nov 15;124(1):43–48. [PubMed]
  • 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. [PMC free article] [PubMed]
  • 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. [PubMed]
  • Kirszbaum L, Sotiropoulos C, Jackson C, Cleal S, Slakeski N, Reynolds EC. 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. [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • Lantz MS, Allen RD, Vail TA, Switalski LM, Hook M. Specific cell components of Bacteroides gingivalis mediate binding and degradation of human fibrinogen. J Bacteriol. 1991 Jan;173(2):495–504. [PMC free article] [PubMed]
  • Li J, Ellen RP, Hoover CI, Felton JR. Association of proteases of Porphyromonas (Bacteroides) gingivalis with its adhesion to Actinomyces viscosus. J Dent Res. 1991 Feb;70(2):82–86. [PubMed]
  • LOWRY OH, ROSEBROUGH NJ, FARR AL, RANDALL RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed]
  • 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. [PMC free article] [PubMed]
  • Nakayama K, Kadowaki T, Okamoto K, Yamamoto K. Construction and characterization of arginine-specific cysteine proteinase (Arg-gingipain)-deficient mutants of Porphyromonas gingivalis. Evidence for significant contribution of Arg-gingipain to virulence. J Biol Chem. 1995 Oct 6;270(40):23619–23626. [PubMed]
  • Nishikata M, Yoshimura F. Characterization of Porphyromonas (bacteroides) gingivalis hemagglutinin as a protease. Biochem Biophys Res Commun. 1991 Jul 15;178(1):336–342. [PubMed]
  • 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. [PubMed]
  • Ogawa T, Hamada S. Hemagglutinating and chemotactic properties of synthetic peptide segments of fimbrial protein from Porphyromonas gingivalis. Infect Immun. 1994 Aug;62(8):3305–3310. [PMC free article] [PubMed]
  • Okuda K, Yamamoto A, Naito Y, Takazoe I, Slots J, Genco RJ. Purification and properties of hemagglutinin from culture supernatant of Bacteroides gingivalis. Infect Immun. 1986 Dec;54(3):659–665. [PMC free article] [PubMed]
  • Orskov I, Birch-Andersen A, Duguid JP, Stenderup J, Orskov F. An adhesive protein capsule of Escherichia coli. Infect Immun. 1985 Jan;47(1):191–200. [PMC free article] [PubMed]
  • Pavloff N, Potempa J, Pike RN, Prochazka V, Kiefer MC, Travis J, Barr PJ. 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. [PubMed]
  • 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]
  • Potempa J, Pavloff N, Travis J. Porphyromonas gingivalis: a proteinase/gene accounting audit. Trends Microbiol. 1995 Nov;3(11):430–434. [PubMed]
  • 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. [PMC free article] [PubMed]
  • Progulske-Fox A, Tumwasorn S, Holt SC. The expression and function of a Bacteroides gingivalis hemagglutinin gene in Escherichia coli. Oral Microbiol Immunol. 1989 Sep;4(3):121–131. [PubMed]
  • Shah HN, Gharbia SE, Progulske-Fox A, Brocklehurst K. Evidence for independent molecular identity and functional interaction of the haemagglutinin and cysteine proteinase (gingivain) of Porphyromonas gingivalis. J Med Microbiol. 1992 Apr;36(4):239–244. [PubMed]
  • Slots J, Gibbons RJ. Attachment of Bacteroides melaninogenicus subsp. asaccharolyticus to oral surfaces and its possible role in colonization of the mouth and of periodontal pockets. Infect Immun. 1978 Jan;19(1):254–264. [PMC free article] [PubMed]
  • Sojar HT, Lee JY, Bedi GS, Cho MI, Genco RJ. Purification, characterization and immunolocalization of fimbrial protein from Porphyromonas (bacteroides) gingivalis. Biochem Biophys Res Commun. 1991 Mar 15;175(2):713–719. [PubMed]
  • Towbin H, Staehelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. [PMC free article] [PubMed]
  • Yoshimura F, Takahashi K, Nodasaka Y, Suzuki T. Purification and characterization of a novel type of fimbriae from the oral anaerobe Bacteroides gingivalis. J Bacteriol. 1984 Dec;160(3):949–957. [PMC free article] [PubMed]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)


Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...