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. 1986 May 1;163(5):1191–1202. doi: 10.1084/jem.163.5.1191

Localization of protective epitopes of the amino terminus of type 5 streptococcal M protein

PMCID: PMC2188111  PMID: 2422314

Abstract

We have used a set of overlapping chemically synthesized peptides representing the amino terminus of type 5 streptococcal M protein to localize protective, as opposed to nonprotective and tissue- crossreactive epitopes that might be appropriate for vaccine formulations. Rabbit antisera raised against SM5(1-35) reacted in high titer with pep M5 by ELISA and opsonized type 5 streptococci. None of the antisera crossreacted with human heart tissue or myosin. Antisera against SM5(26-35) reacted with SM5(1-35) and pep M5 but failed to opsonize type 5 streptococci. Particle-phase ELISA indicated that SM5(26-35) antibodies were directed against nonprotective determinants of pep M5 that were not exposed on the surface of viable organisms. Opsonization and ELISA inhibition assays showed that, of the SM5(1-35) antibodies that reacted with M5, all were inhibited by SM5(14-35), whereas none was inhibited by SM5(26-35), suggesting that the protective epitopes of SM5(1-35) resided between residues 14 and 26. This was confirmed by subsequent chemical synthesis of this region; SM5(14-26) totally inhibited SM5(1-35) antibodies that reacted with pep M5 in ELISA, and completely inhibited opsonization of type 5 streptococci by SM5(1-35) antibodies. SM5(14-26) evoked high titers of type-specific, opsonic antibodies against type 5 streptococci, confirming the protective immunogenicity of this 13-residue peptide of type 5 M protein.

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Selected References

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  1. Beachey E. H., Campbell G. L., Ofek I. Peptic digestion of streptococcal M protein. II. Extraction of M antigen from group A streptococci with pepsin. Infect Immun. 1974 May;9(5):891–896. doi: 10.1128/iai.9.5.891-896.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Beachey E. H., Seyer J. M., Dale J. B., Hasty D. L. Repeating covalent structure and protective immunogenicity of native and synthetic polypeptide fragments of type 24 streptococcal M protein. Mapping of protective and nonprotective epitopes with monoclonal antibodies. J Biol Chem. 1983 Nov 10;258(21):13250–13257. [PubMed] [Google Scholar]
  3. Beachey E. H., Seyer J. M., Dale J. B., Simpson W. A., Kang A. H. Type-specific protective immunity evoked by synthetic peptide of Streptococcus pyogenes M protein. Nature. 1981 Jul 30;292(5822):457–459. doi: 10.1038/292457a0. [DOI] [PubMed] [Google Scholar]
  4. Beachey E. H., Seyer J. M., Kang A. H. Primary structure of protective antigens of type 24 streptococcal M protein. J Biol Chem. 1980 Jul 10;255(13):6284–6289. [PubMed] [Google Scholar]
  5. Beachey E. H., Seyer J. M., Kang A. H. Repeating covalent structure of streptococcal M protein. Proc Natl Acad Sci U S A. 1978 Jul;75(7):3163–3167. doi: 10.1073/pnas.75.7.3163. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Beachey E. H., Stollerman G. H., Chiang E. Y., Chiang T. M., Seyer J. M., Kang A. H. Purification and properties of M protein extracted from group A streptococci with pepsin: covalent structure of the amino terminal region of type 24 M antigen. J Exp Med. 1977 Jun 1;145(6):1469–1483. doi: 10.1084/jem.145.6.1469. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Beachey E. H., Stollerman G. H., Johnson R. H., Ofek I., Bisno A. L. Human immune response to immunization with a structurally defined polypeptide fragment of streptococcal M protein. J Exp Med. 1979 Oct 1;150(4):862–877. doi: 10.1084/jem.150.4.862. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Beachey E. H., Tartar A., Seyer J. M., Chedid L. Epitope-specific protective immunogenicity of chemically synthesized 13-, 18-, and 23-residue peptide fragments of streptococcal M protein. Proc Natl Acad Sci U S A. 1984 Apr;81(7):2203–2207. doi: 10.1073/pnas.81.7.2203. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Chou P. Y., Fasman G. D. Prediction of the secondary structure of proteins from their amino acid sequence. Adv Enzymol Relat Areas Mol Biol. 1978;47:45–148. doi: 10.1002/9780470122921.ch2. [DOI] [PubMed] [Google Scholar]
  10. Dale J. B., Beachey E. H. Epitopes of streptococcal M proteins shared with cardiac myosin. J Exp Med. 1985 Aug 1;162(2):583–591. doi: 10.1084/jem.162.2.583. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dale J. B., Beachey E. H. Multiple, heart-cross-reactive epitopes of streptococcal M proteins. J Exp Med. 1985 Jan 1;161(1):113–122. doi: 10.1084/jem.161.1.113. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Dale J. B., Beachey E. H. Protective antibody against a peptide fragment of type 5 streptococcal M protein cross-reacts with human heart tissue. Trans Assoc Am Physicians. 1982;95:286–291. [PubMed] [Google Scholar]
  13. Dale J. B., Beachey E. H. Protective antigenic determinant of streptococcal M protein shared with sarcolemmal membrane protein of human heart. J Exp Med. 1982 Oct 1;156(4):1165–1176. doi: 10.1084/jem.156.4.1165. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Dale J. B., Beachey E. H. Unique and common protective epitopes among different serotypes of group A streptococcal M proteins defined with hybridoma antibodies. Infect Immun. 1984 Oct;46(1):267–269. doi: 10.1128/iai.46.1.267-269.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Dale J. B., Ofek I., Beachey E. H. Heterogeneity of type-specific and cross-reactive antigenic determinants within a single M protein of group A streptococci. J Exp Med. 1980 May 1;151(5):1026–1038. doi: 10.1084/jem.151.5.1026. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Dale J. B., Seyer J. M., Beachey E. H. Type-specific immunogenicity of a chemically synthesized peptide fragment of type 5 streptococcal M protein. J Exp Med. 1983 Nov 1;158(5):1727–1732. doi: 10.1084/jem.158.5.1727. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hasty D. L., Beachey E. H., Simpson W. A., Dale J. B. Hybridoma antibodies against protective and nonprotective antigenic determinants of a structurally defined polypeptide fragment of streptococcal M protein. J Exp Med. 1982 Apr 1;155(4):1010–1018. doi: 10.1084/jem.155.4.1010. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. LANCEFIELD R. C. Current knowledge of type-specific M antigens of group A streptococci. J Immunol. 1962 Sep;89:307–313. [PubMed] [Google Scholar]
  19. Manjula B. N., Acharya A. S., Mische S. M., Fairwell T., Fischetti V. A. The complete amino acid sequence of a biologically active 197-residue fragment of M protein isolated from type 5 group A streptococci. J Biol Chem. 1984 Mar 25;259(6):3686–3693. [PubMed] [Google Scholar]
  20. 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]
  21. Manjula B. N., Fischetti V. A. Tropomyosin-like seven residue periodicity in three immunologically distinct streptococal M proteins and its implications for the antiphagocytic property of the molecule. J Exp Med. 1980 Mar 1;151(3):695–708. doi: 10.1084/jem.151.3.695. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Phillips G. N., Jr, Flicker P. F., Cohen C., Manjula B. N., Fischetti V. A. Streptococcal M protein: alpha-helical coiled-coil structure and arrangement on the cell surface. Proc Natl Acad Sci U S A. 1981 Aug;78(8):4689–4693. doi: 10.1073/pnas.78.8.4689. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Rothbard J. B., Fernandez R., Schoolnik G. K. Strain-specific and common epitopes of gonococcal pili. J Exp Med. 1984 Jul 1;160(1):208–221. doi: 10.1084/jem.160.1.208. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Seyer J. M., Kang A. H., Beachey E. H. Primary structural similarities between types 5 and 24 M proteins of Streptococcus pyogenes. Biochem Biophys Res Commun. 1980 Jan 29;92(2):546–553. doi: 10.1016/0006-291x(80)90368-x. [DOI] [PubMed] [Google Scholar]
  25. Tainer J. A., Getzoff E. D., Alexander H., Houghten R. A., Olson A. J., Lerner R. A., Hendrickson W. A. The reactivity of anti-peptide antibodies is a function of the atomic mobility of sites in a protein. Nature. 1984 Nov 8;312(5990):127–134. doi: 10.1038/312127a0. [DOI] [PubMed] [Google Scholar]
  26. Yoshitake S., Yamada Y., Ishikawa E., Masseyeff R. Conjugation of glucose oxidase from Aspergillus niger and rabbit antibodies using N-hydroxysuccinimide ester of N-(4-carboxycyclohexylmethyl)-maleimide. Eur J Biochem. 1979 Nov;101(2):395–399. doi: 10.1111/j.1432-1033.1979.tb19731.x. [DOI] [PubMed] [Google Scholar]

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