Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1989 Dec;57(12):3834–3840. doi: 10.1128/iai.57.12.3834-3840.1989

Purification of a Streptococcus mutans protein that binds to heart tissue and glycosaminoglycans.

S H Choi 1, M W Stinson 1
PMCID: PMC259913  PMID: 2680984

Abstract

Proteins of Streptococcus mutans MT703 were isolated by differential filtration from chemically defined culture medium following growth of the bacteria. Incubation of this preparation with cryostat-cut sections of fresh rabbit cardiac muscle resulted in deposition of streptococcal components on basement membranes of sarcolemmal sheaths and capillary walls, as indicated by indirect immunofluorescence assay. Binding of radioiodinated streptococcal proteins to heart in vitro was time dependent and saturable. Unlabeled S. mutans proteins competitively inhibited 72% of heart binding by the radiolabeled proteins, indicating a high level of binding specificity. A selection of components common to tissue basement membranes was tested for their abilities to inhibit the binding of streptococcal proteins to heart tissue. Of the glycosaminoglycans, heparin was the most effective inhibitor, followed by heparan sulfate and chondroitin sulfate. Hyaluronic acid was not inhibitory. Of the glycoproteins tested, laminin and collagen type IV were weakly inhibitory, whereas fibronectin was ineffective. A single polypeptide was purified to homogeneity by affinity chromatography on a column of heparin-agarose. Gel filtration chromatography of the purified protein under nondissociating conditions showed a single component at 31 kilodaltons (kDa), whereas in sodium dodecyl sulfate-polyacrylamide gel electrophoresis one band appeared at 8 kDa. This indicates that the tissue-binding protein may either be a linear polypeptide or be released into the environment by the bacterium as a tetramer of the 8-kDa polypeptide. The purified protein had an isoelectric point of 9.5 and showed binding activity for basement membranes in thin sections of heart. Chemical analyses of the purified binding protein showed it to have high contents of lysine and alanine and to be devoid of half-cystine, methionine, tyrosine, histidine, and both neutral and amino sugars.

Full text

PDF
3834

Images in this article

3837Image
on p.3837
3838Image
on p.3838

Selected References

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

  1. Albini B., Nisengard R. J., Glurich I., Neiders M. E., Stinson M. W. Streptococcus mutans-induced nephritis in rabbits. Am J Pathol. 1985 Mar;118(3):408–418. [PMC free article] [PubMed] [Google Scholar]
  2. Bergey E. J., Stinson M. W. Heparin-inhibitable basement membrane-binding protein of Streptococcus pyogenes. Infect Immun. 1988 Jul;56(7):1715–1721. doi: 10.1128/iai.56.7.1715-1721.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Gallagher J. T., Lyon M., Steward W. P. Structure and function of heparan sulphate proteoglycans. Biochem J. 1986 Jun 1;236(2):313–325. doi: 10.1042/bj2360313. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Gibbons R. J., van Houte J. Dental caries. Annu Rev Med. 1975;26:121–136. doi: 10.1146/annurev.me.26.020175.001005. [DOI] [PubMed] [Google Scholar]
  5. Gutman R. A., Striker G. E., Gilliland B. C., Cutler R. E. The immune complex glomerulonephritis of bacterial endocarditis. Medicine (Baltimore) 1972 Jan;51(1):1–25. doi: 10.1097/00005792-197201000-00001. [DOI] [PubMed] [Google Scholar]
  6. Harder E. J., Wilkowske C. J., Washington J. A., 2nd, Geraci J. E. Streptococcus mutans endocarditis. Ann Intern Med. 1974 Mar;80(3):364–368. doi: 10.7326/0003-4819-80-3-364. [DOI] [PubMed] [Google Scholar]
  7. Hughes M., Machardy S. M., Sheppard A. J., Woods N. C. Evidence for an immunological relationship between Streptococcus mutans and human cardiac tissue. Infect Immun. 1980 Feb;27(2):576–588. doi: 10.1128/iai.27.2.576-588.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Kanwar Y. S., Linker A., Farquhar M. G. Increased permeability of the glomerular basement membrane to ferritin after removal of glycosaminoglycans (heparan sulfate) by enzyme digestion. J Cell Biol. 1980 Aug;86(2):688–693. doi: 10.1083/jcb.86.2.688. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. Loesche W. J. Role of Streptococcus mutans in human dental decay. Microbiol Rev. 1986 Dec;50(4):353–380. doi: 10.1128/mr.50.4.353-380.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. McGhee J. R., Michalek S. M. Immunobiology of dental caries: microbial aspects and local immunity. Annu Rev Microbiol. 1981;35:595–638. doi: 10.1146/annurev.mi.35.100181.003115. [DOI] [PubMed] [Google Scholar]
  12. Morrissey J. H. Silver stain for proteins in polyacrylamide gels: a modified procedure with enhanced uniform sensitivity. Anal Biochem. 1981 Nov 1;117(2):307–310. doi: 10.1016/0003-2697(81)90783-1. [DOI] [PubMed] [Google Scholar]
  13. Obrind B., Pertoft H., Iverius P. H., Laurent The effect of calcium on the macromolecular properties of heparan sulfate. Connect Tissue Res. 1975;3(2):187–193. doi: 10.3109/03008207509152178. [DOI] [PubMed] [Google Scholar]
  14. Roberts R. B., Krieger A. G., Schiller N. L., Gross K. C. Viridans streptococcal endocarditis: the role of various species, including pyridoxal-dependent streptococci. Rev Infect Dis. 1979 Nov-Dec;1(6):955–966. doi: 10.1093/clinids/1.6.955. [DOI] [PubMed] [Google Scholar]
  15. Seiler M. W., Venkatachalam M. A., Cotran R. S. Glomerular epithelium: structural alterations induced by polycations. Science. 1975 Aug 1;189(4200):390–393. doi: 10.1126/science.1145209. [DOI] [PubMed] [Google Scholar]
  16. Stinson M. W., Barua P. K., Bergey E. J., Nisengard R. J., Neiders M. E., Albini B. Binding of Streptococcus mutans antigens to heart and kidney basement membranes. Infect Immun. 1984 Oct;46(1):145–151. doi: 10.1128/iai.46.1.145-151.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Stinson M. W., Nisengard R. J., Neiders M. E., Albini B. Serology and tissue lesions in rabbits immunized with Streptococcus mutans. J Immunol. 1983 Dec;131(6):3021–3027. [PubMed] [Google Scholar]
  18. Terleckyj B., Willett N. P., Shockman G. D. Growth of several cariogenic strains of oral streptococci in a chemically defined medium. Infect Immun. 1975 Apr;11(4):649–655. doi: 10.1128/iai.11.4.649-655.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES