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Infection and Immunity logoLink to Infection and Immunity
. 1975 Sep;12(3):679–686. doi: 10.1128/iai.12.3.679-686.1975

Guanidine extraction of streptococcal M protein.

H Russell, R R Facklam
PMCID: PMC415339  PMID: 809363

Abstract

A new method of extracting M protein from streptococcal cell walls has been presented. The extracting agent was guanidine-hydrochloride, a protein denaturant. The crude guanidine extract was further purified by ammonium sulfate and pH 5 fractionation and by hydroxyapatite column chromatography. Three major protein peaks were eluted from the hydroxyapatite column with 0.01, 0.1 and 0.3 M phosphate buffer, respectively. Protein fractions eluted at 0.1 and 0.3 M phosphate concentractions contained antigens that precipitated with homologous M-protein specific antisera, whereas the 0.01 M phosphate fraction had no immunological activity. The fraction eluted with 0.3 M phosphate was electrophoretically homogeneous in sodium dodecyl sulfate-acrylamide gels and elicited the production of bactericidal antibodies in rabbits. The 0.1 M phosphate buffer eluant was electrophoretically heterogeneous and did not elicit the production of bactericidal antibodies in rabbits.

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Selected References

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  1. BECKER C. G. SELECTION OF GROUP A STREPTOCOCCI RICH IN M-PROTEIN FROM POPULATIONS POOR IN M-PROTEIN. Am J Pathol. 1964 Jan;44:51–60. [PMC free article] [PubMed] [Google Scholar]
  2. BLEIWEIS A. S., KARAKAWA W. W., KRAUSE R. M. IMPROVED TECHNIQUE FOR THE PREPARATION OF STREPTOCOCCAL CELL WALLS. J Bacteriol. 1964 Oct;88:1198–1200. doi: 10.1128/jb.88.4.1198-1200.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Beachey E. H., Cunningham M. Type-specific inhibition of preopsonization versus immunoprecipitation by Streptococcal M proteins. Infect Immun. 1973 Jul;8(1):19–24. doi: 10.1128/iai.8.1.19-24.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. DAVIS B. J. DISC ELECTROPHORESIS. II. METHOD AND APPLICATION TO HUMAN SERUM PROTEINS. Ann N Y Acad Sci. 1964 Dec 28;121:404–427. doi: 10.1111/j.1749-6632.1964.tb14213.x. [DOI] [PubMed] [Google Scholar]
  5. FOX E. N. ANTIGENICITY OF THE M PROTEINS OF GROUP A HEMOLYTIC STREPTOCOCCI. J Immunol. 1964 Nov;93:826–837. [PubMed] [Google Scholar]
  6. Fox E. N. M proteins of group A streptococci. Bacteriol Rev. 1974 Mar;38(1):57–86. doi: 10.1128/br.38.1.57-86.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fox E. N., Wittner M. K. Antigenicity of the M proteins of group A hemolytic streptococci. IV. Cross-reactivity between serotypes. J Immunol. 1968 Jan;100(1):39–45. [PubMed] [Google Scholar]
  8. Ginsburg I. Mechanisms of cell and tissue injury induced by group A streptococci: relation to poststreptococcal sequelae. J Infect Dis. 1972 Oct;126(4):419–456. doi: 10.1093/infdis/126.4.419. [DOI] [PubMed] [Google Scholar]
  9. Johnson R. H., Vosti K. L. Purification of two fragments of M protein from a strain of group A, type 12 streptococcus. J Immunol. 1968 Sep;101(3):381–391. [PubMed] [Google Scholar]
  10. LANCEFIELD R. C. Current knowledge of type-specific M antigens of group A streptococci. J Immunol. 1962 Sep;89:307–313. [PubMed] [Google Scholar]
  11. LANCEFIELD R. C. Differentiation of group A streptococci with a common R antigen into three serological types, with special reference to the bactericidal test. J Exp Med. 1957 Oct 1;106(4):525–544. doi: 10.1084/jem.106.4.525. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. LANCEFIELD R. C., PERLMANN G. E. Preparation and properties of type-specific M antigen isolated from a group A, type 1 hemolytic streptococcus. J Exp Med. 1952 Jul;96(1):71–82. doi: 10.1084/jem.96.1.71. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  14. Ofek I., Bergner-Rabinowitz S., Davies A. M. Opsonic capacity of type specific streptococcal antibodies. Isr J Med Sci. 1969 May-Jun;5(3):293–296. [PubMed] [Google Scholar]
  15. Vosti K. L., Johnson R. H., Dillon M. F. Further characterization of purified fractions of M protein from a strain of group A, type 12 Streptococcus. J Immunol. 1971 Jul;107(1):104–114. [PubMed] [Google Scholar]
  16. WADSWORTH C. A slide microtechnique for the analysis of immune precipitates in gel. Int Arch Allergy Appl Immunol. 1957;10(6):355–360. doi: 10.1159/000228394. [DOI] [PubMed] [Google Scholar]
  17. WHITE F. H., Jr Regeneration of native secondary and tertiary structures by air oxidation of reduced ribonuclease. J Biol Chem. 1961 May;236:1353–1360. [PubMed] [Google Scholar]

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