Skip to main content
Infection and Immunity logoLink to Infection and Immunity
. 1988 Sep;56(9):2417–2423. doi: 10.1128/iai.56.9.2417-2423.1988

Characterization of an adhesion antigen of Streptococcus sanguis G9B.

R J Lamont 1, B Rosan 1, C T Baker 1, G M Nelson 1
PMCID: PMC259582  PMID: 2842262

Abstract

An antigen possessing the attributes of an adhesion has been identified in Streptococcus sanguis G9B. Cell surface components were extracted from G9B and a spontaneously occurring nonadherent mutant of G9B, strain Adh-, with a 2 mM barbital buffer, pH 8.6. The extract of G9B but not of Adh- absorbed more than 80% of the adhesion-inhibitory activity of anti-G9B immunoglobulin G (IgG). Immunoblots revealed 80- and 52-kilodalton (kDa) antigens present in the G9B extract but not in the Adh- extract. Absorption of anti-G9B IgG with Adh- and G9B barbital extracts showed a correlation between the loss of the 80- and 52-kDa antibodies and the loss of adhesion-inhibitory activity. An antibody prepared against the 80-kDa antigen excised from sodium dodecyl sulfate-polyacrylamide gels recognized the 80- and 52-kDa antigens and another antigen of 62 kDa but did not inhibit adhesion. However, an antibody from an electroblot containing the native protein from which the 80-kDa and related antigens were derived (the 80-kDa antigen complex) inhibited adhesion to the same extent as anti-G9B IgG. Periodate oxidation of the G9B barbital extract modified the 80-kDa antigen complex and resulted in the loss of 40% of its absorbing activity. The barbital extract also contained an endogenous enzyme responsible for producing the 62- and 52-kDa antigens from the 80-kDa protein and which, under optimal conditions, degraded the antigen completely, resulting in the loss of antibody-absorbing activity. The 80-kDa antigen complex has a molecular mass of more than 200 kDa in native polyacrylamide gels and a pI of 4.1 to 4.8. These observations suggest that the adhesion antigen in S. sanguis G9B is a large glycoprotein from which an 80-kDa antigen complex is derived.

Full text

PDF
2417

Images in this article

Selected References

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

  1. Ames G. F., Nikaido K. Two-dimensional gel electrophoresis of membrane proteins. Biochemistry. 1976 Feb 10;15(3):616–623. doi: 10.1021/bi00648a026. [DOI] [PubMed] [Google Scholar]
  2. Appelbaum B., Rosan B. Cell surface proteins of oral streptococci. Infect Immun. 1984 Oct;46(1):245–250. doi: 10.1128/iai.46.1.245-250.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bergey E. J., Levine M. J., Reddy M. S., Bradway S. D., Al-Hashimi I. Use of the photoaffinity cross-linking agent N-hydroxysuccinimidyl-4-azidosalicylic acid to characterize salivary-glycoprotein-bacterial interactions. Biochem J. 1986 Feb 15;234(1):43–48. doi: 10.1042/bj2340043. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  5. Cisar J. O., Curl S. H., Kolenbrander P. E., Vatter A. E. Specific absence of type 2 fimbriae on a coaggregation-defective mutant of Actinomyces viscosus T14V. Infect Immun. 1983 May;40(2):759–765. doi: 10.1128/iai.40.2.759-765.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cowan M. M., Taylor K. G., Doyle R. J. Energetics of the initial phase of adhesion of Streptococcus sanguis to hydroxylapatite. J Bacteriol. 1987 Jul;169(7):2995–3000. doi: 10.1128/jb.169.7.2995-3000.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cowan M. M., Taylor K. G., Doyle R. J. Kinetic analysis of Streptococcus sanguis adhesion to artificial pellicle. J Dent Res. 1986 Oct;65(10):1278–1283. doi: 10.1177/00220345860650101501. [DOI] [PubMed] [Google Scholar]
  8. Eifert R., Rosan B., Golub E. Optimization of an hydroxyapatite adhesion assay for Streptococcus sanguis. Infect Immun. 1984 May;44(2):287–291. doi: 10.1128/iai.44.2.287-291.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Elder B. L., Fives-Taylor P. Characterization of monoclonal antibodies specific for adhesion: isolation of an adhesin of Streptococcus sanguis FW213. Infect Immun. 1986 Nov;54(2):421–427. doi: 10.1128/iai.54.2.421-427.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fachon-Kalweit S., Elder B. L., Fives-Taylor P. Antibodies that bind to fimbriae block adhesion of Streptococcus sanguis to saliva-coated hydroxyapatite. Infect Immun. 1985 Jun;48(3):617–624. doi: 10.1128/iai.48.3.617-624.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Facklam R. R. Physiological differentiation of viridans streptococci. J Clin Microbiol. 1977 Feb;5(2):184–201. doi: 10.1128/jcm.5.2.184-201.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fives-Taylor P. M., Thompson D. W. Surface properties of Streptococcus sanguis FW213 mutants nonadherent to saliva-coated hydroxyapatite. Infect Immun. 1985 Mar;47(3):752–759. doi: 10.1128/iai.47.3.752-759.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gibbons R. J., Etherden I. Albumin as a blocking agent in studies of streptococcal adsorption to experimental salivary pellicles. Infect Immun. 1985 Nov;50(2):592–594. doi: 10.1128/iai.50.2.592-594.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gibbons R. J., Hay D. I. Human salivary acidic proline-rich proteins and statherin promote the attachment of Actinomyces viscosus LY7 to apatitic surfaces. Infect Immun. 1988 Feb;56(2):439–445. doi: 10.1128/iai.56.2.439-445.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gibbons R. J., Moreno E. C., Etherden I. Concentration-dependent multiple binding sites on saliva-treated hydroxyapatite for Streptococcus sanguis. Infect Immun. 1983 Jan;39(1):280–289. doi: 10.1128/iai.39.1.280-289.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Gibbons R. J., van Houte J. Selective bacterial adherence to oral epithelial surfaces and its role as an ecological determinant. Infect Immun. 1971 Apr;3(4):567–573. doi: 10.1128/iai.3.4.567-573.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hamada S., Slade H. D. Biology, immunology, and cariogenicity of Streptococcus mutans. Microbiol Rev. 1980 Jun;44(2):331–384. doi: 10.1128/mr.44.2.331-384.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. Lamont R. J., Rosan B., Murphy G. M., Baker C. T. Streptococcus sanguis surface antigens and their interactions with saliva. Infect Immun. 1988 Jan;56(1):64–70. doi: 10.1128/iai.56.1.64-70.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Levine M. J., Herzberg M. C., Levine M. S., Ellison S. A., Stinson M. W., Li H. C., van Dyke T. Specificity of salivary-bacterial interactions: role of terminal sialic acid residues in the interaction of salivary glycoproteins with Streptococcus sanguis and Streptococcus mutans. Infect Immun. 1978 Jan;19(1):107–115. doi: 10.1128/iai.19.1.107-115.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Liljemark W. F., Bloomquist C. G. Isolation of a protein-containing cell surface component from Streptococcus sanguis which affects its adherence to saliva-coated hydroxyapatite. Infect Immun. 1981 Nov;34(2):428–434. doi: 10.1128/iai.34.2.428-434.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. McBride B. C., Song M., Krasse B., Olsson J. Biochemical and immunological differences between hydrophobic and hydrophilic strains of Streptococcus mutans. Infect Immun. 1984 Apr;44(1):68–75. doi: 10.1128/iai.44.1.68-75.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Mitchell C. G., Smith R., Lehner T. Recognition of carbohydrate and protein epitopes by monoclonal antibodies to a cell wall antigen from Streptococcus mutans. Infect Immun. 1987 Mar;55(3):810–815. doi: 10.1128/iai.55.3.810-815.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Morris E. J., Ganeshkumar N., McBride B. C. Cell surface components of Streptococcus sanguis: relationship to aggregation, adherence, and hydrophobicity. J Bacteriol. 1985 Oct;164(1):255–262. doi: 10.1128/jb.164.1.255-262.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Morris E. J., Ganeshkumar N., Song M., McBride B. C. Identification and preliminary characterization of a Streptococcus sanguis fibrillar glycoprotein. J Bacteriol. 1987 Jan;169(1):164–171. doi: 10.1128/jb.169.1.164-171.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Morris E. J., McBride B. C. Adherence of Streptococcus sanguis to saliva-coated hydroxyapatite: evidence for two binding sites. Infect Immun. 1984 Feb;43(2):656–663. doi: 10.1128/iai.43.2.656-663.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Nesbitt W. E., Doyle R. J., Taylor K. G., Staat R. H., Arnold R. R. Positive coooperativity in the binding of Streptococcus sanguis to hydroxylapatite. Infect Immun. 1982 Jan;35(1):157–165. doi: 10.1128/iai.35.1.157-165.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Olmsted J. B. Affinity purification of antibodies from diazotized paper blots of heterogeneous protein samples. J Biol Chem. 1981 Dec 10;256(23):11955–11957. [PubMed] [Google Scholar]
  29. Reinhart M. P., Malamud D. Protein transfer from isoelectric focusing Gels: the native blot. Anal Biochem. 1982 Jul 1;123(2):229–235. doi: 10.1016/0003-2697(82)90439-0. [DOI] [PubMed] [Google Scholar]
  30. Rosan B. A comparison of the phenol water and Rantz and Randall teichoic acid antigens in group H streptococci. Adv Exp Med Biol. 1978;107:791–802. doi: 10.1007/978-1-4684-3369-2_89. [DOI] [PubMed] [Google Scholar]
  31. Rosan B. Antigens of Streptococcus sanguis. Infect Immun. 1973 Feb;7(2):205–211. doi: 10.1128/iai.7.2.205-211.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Rosan B., Appelbaum B., Campbell L. K., Knox K. W., Wicken A. J. Chemostat studies of the effect of environmental control on Streptococcus sanguis adherence to hydroxyapatite. Infect Immun. 1982 Jan;35(1):64–70. doi: 10.1128/iai.35.1.64-70.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Rosan B. Relationship of the cell wall composition of group H streptococci and Streptococcus sanguis to their serological properties. Infect Immun. 1976 Apr;13(4):1144–1153. doi: 10.1128/iai.13.4.1144-1153.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. 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. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Weerkamp A. H., Handley P. S., Baars A., Slot J. W. Negative staining and immunoelectron microscopy of adhesion-deficient mutants of Streptococcus salivarius reveal that the adhesive protein antigens are separate classes of cell surface fibril. J Bacteriol. 1986 Mar;165(3):746–755. doi: 10.1128/jb.165.3.746-755.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Wyatt J. E., Hesketh L. M., Handley P. S. Lack of correlation between fibrils, hydrophobicity and adhesion for strains of Streptococcus sanguis biotypes I and II. Microbios. 1987;50(202):7–15. [PubMed] [Google Scholar]

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

RESOURCES