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. 1994 Feb;62(2):442–448. doi: 10.1128/iai.62.2.442-448.1994

M protein mediates streptococcal adhesion to HEp-2 cells.

J R Wang 1, M W Stinson 1
PMCID: PMC186127  PMID: 8300205

Abstract

Streptococcus pyogenes adheres to human epithelial cells in vitro and in vivo. To identify adhesins, cell wall components were extracted from S. pyogenes M6 with alkali or by treatment with mutanolysin and lysozyme. HEp-2 cells were incubated with extracts of S. pyogenes M6 and then analyzed by Western blot (immunoblot) assays, using antibodies to S. pyogenes. Only one streptococcal component (62 kDa) was bound to HEp-2 cells and was identified serologically as M6 protein. Experiments with pepsin-cleaved fragments of M protein indicated that the binding site was located at the N-terminal half of the molecule. M protein was bound selectively to two trypsin-sensitive surface components, 97 and 205 kDa, of HEp-2 cells on nitrocellulose blots of sodium dodecyl sulfate-polyacrylamide gels. Tritium-labeled lipoteichoic acid bound to different HEp-2 cell components, 34 and 35 kDa, in a parallel experiment, indicating that lipoteichoic acid was not complexed with M protein and does not mediate M-protein binding. The four HEp-2 components were unrelated to fibronectin since they did not react with specific antibodies. An M-protein-deficient (M-) strain of streptococcus (JRS75), grown in chemically defined medium, showed 73% less adhesion activity to HEp-2 monolayers than an M+ strain (JRS4). Streptococcal adhesion was insensitive to competitive inhibition by selected monosaccharides. These results indicate that M protein binds directly to certain HEp-2 cell membrane components and mediates streptococcal adhesion.

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Selected References

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

  1. Beachey E. H. Bacterial adherence: adhesin-receptor interactions mediating the attachment of bacteria to mucosal surface. J Infect Dis. 1981 Mar;143(3):325–345. doi: 10.1093/infdis/143.3.325. [DOI] [PubMed] [Google Scholar]
  2. Caparon M. G., Stephens D. S., Olsén A., Scott J. R. Role of M protein in adherence of group A streptococci. Infect Immun. 1991 May;59(5):1811–1817. doi: 10.1128/iai.59.5.1811-1817.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Courtney H. S., Hasty D. L., Dale J. B., Poirier T. P. A 28-kilodalton fibronectin-binding protein of group A streptococci. Curr Microbiol. 1992 Nov;25(5):245–250. doi: 10.1007/BF01575856. [DOI] [PubMed] [Google Scholar]
  4. Courtney H. S., von Hunolstein C., Dale J. B., Bronze M. S., Beachey E. H., Hasty D. L. Lipoteichoic acid and M protein: dual adhesins of group A streptococci. Microb Pathog. 1992 Mar;12(3):199–208. doi: 10.1016/0882-4010(92)90054-r. [DOI] [PubMed] [Google Scholar]
  5. Ellen R. P., Gibbons R. J. M protein-associated adherence of Streptococcus pyogenes to epithelial surfaces: prerequisite for virulence. Infect Immun. 1972 May;5(5):826–830. doi: 10.1128/iai.5.5.826-830.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Ellen R. P., Gibbons R. J. Parameters affecting the adherence and tissue tropisms of Streptococcus pyogenes. Infect Immun. 1974 Jan;9(1):85–91. doi: 10.1128/iai.9.1.85-91.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fischetti V. A., Jones K. F., Manjula B. N., Scott J. R. Streptococcal M6 protein expressed in Escherichia coli. Localization, purification, and comparison with streptococcal-derived M protein. J Exp Med. 1984 Apr 1;159(4):1083–1095. doi: 10.1084/jem.159.4.1083. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Fischetti V. A., Parry D. A., Trus B. L., Hollingshead S. K., Scott J. R., Manjula B. N. Conformational characteristics of the complete sequence of group A streptococcal M6 protein. Proteins. 1988;3(1):60–69. doi: 10.1002/prot.340030106. [DOI] [PubMed] [Google Scholar]
  9. Fischetti V. A. Streptococcal M protein: molecular design and biological behavior. Clin Microbiol Rev. 1989 Jul;2(3):285–314. doi: 10.1128/cmr.2.3.285. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gibbons R. J., Dankers I. Association of food lectins with human oral epithelial cells in vivo. Arch Oral Biol. 1983;28(7):561–566. doi: 10.1016/0003-9969(83)90002-x. [DOI] [PubMed] [Google Scholar]
  11. Gibbons R. J., Houte J. V. Bacterial adherence in oral microbial ecology. Annu Rev Microbiol. 1975;29:19–44. doi: 10.1146/annurev.mi.29.100175.000315. [DOI] [PubMed] [Google Scholar]
  12. Grabovskaya K. B., Totoljan A. A., Rýc M., Havlícek J., Burova L. A., Bícová R. Adherence of group A streptococci to epithelial cells in tissue culture. Zentralbl Bakteriol A. 1980 Aug;247(3):303–314. [PubMed] [Google Scholar]
  13. Hanski E., Caparon M. Protein F, a fibronectin-binding protein, is an adhesin of the group A streptococcus Streptococcus pyogenes. Proc Natl Acad Sci U S A. 1992 Jul 1;89(13):6172–6176. doi: 10.1073/pnas.89.13.6172. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hasty D. L., Ofek I., Courtney H. S., Doyle R. J. Multiple adhesins of streptococci. Infect Immun. 1992 Jun;60(6):2147–2152. doi: 10.1128/iai.60.6.2147-2152.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hasty D. L., Simpson W. A. Effects of fibronectin and other salivary macromolecules on the adherence of Escherichia coli to buccal epithelial cells. Infect Immun. 1987 Sep;55(9):2103–2109. doi: 10.1128/iai.55.9.2103-2109.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hollingshead S. K., Fischetti V. A., Scott J. R. Complete nucleotide sequence of type 6 M protein of the group A Streptococcus. Repetitive structure and membrane anchor. J Biol Chem. 1986 Feb 5;261(4):1677–1686. [PubMed] [Google Scholar]
  17. Hollingshead S. K., Simecka J. W., Michalek S. M. Role of M protein in pharyngeal colonization by group A streptococci in rats. Infect Immun. 1993 Jun;61(6):2277–2283. doi: 10.1128/iai.61.6.2277-2283.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Horstmann R. D., Sievertsen H. J., Knobloch J., Fischetti V. A. Antiphagocytic activity of streptococcal M protein: selective binding of complement control protein factor H. Proc Natl Acad Sci U S A. 1988 Mar;85(5):1657–1661. doi: 10.1073/pnas.85.5.1657. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hyzy J., Sciotti V., Albini B., Stinson M. Deposition of circulating streptococcal lipoteichoic acid in mouse tissues. Microb Pathog. 1992 Aug;13(2):123–132. doi: 10.1016/0882-4010(92)90072-v. [DOI] [PubMed] [Google Scholar]
  20. Isberg R. R., Falkow S. A single genetic locus encoded by Yersinia pseudotuberculosis permits invasion of cultured animal cells by Escherichia coli K-12. Nature. 1985 Sep 19;317(6034):262–264. doi: 10.1038/317262a0. [DOI] [PubMed] [Google Scholar]
  21. Jones K. F., Fischetti V. A. The importance of the location of antibody binding on the M6 protein for opsonization and phagocytosis of group A M6 streptococci. J Exp Med. 1988 Mar 1;167(3):1114–1123. doi: 10.1084/jem.167.3.1114. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Jones K. F., Hollingshead S. K., Scott J. R., Fischetti V. A. Spontaneous M6 protein size mutants of group A streptococci display variation in antigenic and opsonogenic epitopes. Proc Natl Acad Sci U S A. 1988 Nov;85(21):8271–8275. doi: 10.1073/pnas.85.21.8271. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Jones K. F., Khan S. A., Erickson B. W., Hollingshead S. K., Scott J. R., Fischetti V. A. Immunochemical localization and amino acid sequences of crossreactive epitopes within the group A streptococcal M6 protein. J Exp Med. 1986 Oct 1;164(4):1226–1238. doi: 10.1084/jem.164.4.1226. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Jones K. F., Manjula B. N., Johnston K. H., Hollingshead S. K., Scott J. R., Fischetti V. A. Location of variable and conserved epitopes among the multiple serotypes of streptococcal M protein. J Exp Med. 1985 Mar 1;161(3):623–628. doi: 10.1084/jem.161.3.623. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Knipe D. M., Baltimore D., Lodish H. F. Separate pathways of maturation of the major structural proteins of vesicular stomatitis virus. J Virol. 1977 Mar;21(3):1128–1139. doi: 10.1128/jvi.21.3.1128-1139.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. LANCEFIELD R. C. Current knowledge of type-specific M antigens of group A streptococci. J Immunol. 1962 Sep;89:307–313. [PubMed] [Google Scholar]
  27. 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]
  28. Manjula B. N., Fischetti V. A. Studies on group A streptococcal M-proteins: purification of type 5 M-protein and comparison of its amino terminal sequence with two immunologically unrelated M-protein molecules. J Immunol. 1980 Jan;124(1):261–267. [PubMed] [Google Scholar]
  29. Norgren M., Caparon M. G., Scott J. R. A method for allelic replacement that uses the conjugative transposon Tn916: deletion of the emm6.1 allele in Streptococcus pyogenes JRS4. Infect Immun. 1989 Dec;57(12):3846–3850. doi: 10.1128/iai.57.12.3846-3850.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Ofek I., Simpson W. A., Beachey E. H. Formation of molecular complexes between a structurally defined M protein and acylated or deacylated lipoteichoic acid of Streptococcus pyogenes. J Bacteriol. 1982 Feb;149(2):426–433. doi: 10.1128/jb.149.2.426-433.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Scott J. R., Fischetti V. A. Expression of streptococcal M protein in Escherichia coli. Science. 1983 Aug 19;221(4612):758–760. doi: 10.1126/science.6192499. [DOI] [PubMed] [Google Scholar]
  32. Scott J. R., Guenthner P. C., Malone L. M., Fischetti V. A. Conversion of an M- group A streptococcus to M+ by transfer of a plasmid containing an M6 gene. J Exp Med. 1986 Nov 1;164(5):1641–1651. doi: 10.1084/jem.164.5.1641. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Selinger D. S., Julie N., Reed W. P., Williams R. C., Jr Adherence of group A streptococci to pharyngeal cells: a role in the pathogenesis of rheumatic fever. Science. 1978 Aug 4;201(4354):455–457. doi: 10.1126/science.351810. [DOI] [PubMed] [Google Scholar]
  34. Simpson W. A., Courtney H. S., Ofek I. Interactions of fibronectin with streptococci: the role of fibronectin as a receptor for Streptococcus pyogenes. Rev Infect Dis. 1987 Jul-Aug;9 (Suppl 4):S351–S359. doi: 10.1093/clinids/9.supplement_4.s351. [DOI] [PubMed] [Google Scholar]
  35. Simpson W. A., Ofek I., Beachey E. H. Fatty acid binding sites of serum albumin as membrane receptor analogs for streptococcal lipoteichoic acid. Infect Immun. 1980 Jul;29(1):119–122. doi: 10.1128/iai.29.1.119-122.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Stinson M. W., Jones C. A. Binding of Todd-Hewitt broth antigens by Streptococcus mutans. Infect Immun. 1983 Jun;40(3):1140–1145. doi: 10.1128/iai.40.3.1140-1145.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Talay S. R., Valentin-Weigand P., Jerlström P. G., Timmis K. N., Chhatwal G. S. Fibronectin-binding protein of Streptococcus pyogenes: sequence of the binding domain involved in adherence of streptococci to epithelial cells. Infect Immun. 1992 Sep;60(9):3837–3844. doi: 10.1128/iai.60.9.3837-3844.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. 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]
  39. Whitnack E., Beachey E. H. Biochemical and biological properties of the binding of human fibrinogen to M protein in group A streptococci. J Bacteriol. 1985 Oct;164(1):350–358. doi: 10.1128/jb.164.1.350-358.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Yu C. E., Ferretti J. J. Molecular epidemiologic analysis of the type A streptococcal exotoxin (erythrogenic toxin) gene (speA) in clinical Streptococcus pyogenes strains. Infect Immun. 1989 Dec;57(12):3715–3719. doi: 10.1128/iai.57.12.3715-3719.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. van de Rijn I., Kessler R. E. Growth characteristics of group A streptococci in a new chemically defined medium. Infect Immun. 1980 Feb;27(2):444–448. doi: 10.1128/iai.27.2.444-448.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]

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