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. 1974 Jan;27(1):28–37. doi: 10.1128/am.27.1.28-37.1974

Simplified Method for the Purification of Group A Streptococcal M-Proteins: Solution of the Multiple Banding Problem

David C Straus 1, Aruna Mehta 1, Charles F Lange 1
PMCID: PMC379963  PMID: 4203786

Abstract

A simple and rapid procedure for the isolation in high yield (about a 30% recovery based on the total 30 to 60% ammonium sulfate recovery) of homogeneous purified group A streptococcal M-protein is described. M-proteins extracted from whole cells of group A streptococci by treatment with hot HCl were neutralized, fractionated with ammonium sulfate, dialyzed, lyophilized, and then subjected to treatment with hot 60% trichloroacetic acid. This was shown to produce an M-protein preparation, free of group A carbohydrate activity and extraneous antigens, in yields up to 10-fold higher than previous methods in about one-fifth the time. These M-protein preparations were shown to: (i) have similar amino acid compositions to their respective type-specific proteins purified by diethylaminoethyl and O-(carboxymethyl) cellulose chromatography, (ii) react with their respective type-specific antisera in Ouchterlony diffusion, (iii) produce antisera in rabbits capable of promoting streptococcal long-chain formation in vitro, and (iv) give only one major band on polyacrylamide gel disk electrophoresis. The data allow for an explanation of the hitherto described multiple banding M-proteins seen on acrylamide electrophoresis.

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Selected References

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  1. CLARK H. F., SHEPARD C. C. A DIALYSIS TECHNIQUE FOR PREPARING FLUORESCENT ANTIBODY. Virology. 1963 Aug;20:642–644. doi: 10.1016/0042-6822(63)90292-7. [DOI] [PubMed] [Google Scholar]
  2. FRAENKEL-CONRAT H., PORTER R. R. The terminal amino groups of conalbumin, ovomucoid and avidin. Biochim Biophys Acta. 1952 Nov;9(5):557–562. [PubMed] [Google Scholar]
  3. Fox E. N., Wittner M. K. New observations on the structure and antigenicity of the M proteins of the group A streptococcus. Immunochemistry. 1969 Jan;6(1):11–24. doi: 10.1016/0019-2791(69)90174-8. [DOI] [PubMed] [Google Scholar]
  4. Fox E. N., Wittner M. K. The multiple molecular structure of the M proteins of group A streptococci. Proc Natl Acad Sci U S A. 1965 Oct;54(4):1118–1125. doi: 10.1073/pnas.54.4.1118. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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]
  6. LANCEFIELD R. C. Current knowledge of type-specific M antigens of group A streptococci. J Immunol. 1962 Sep;89:307–313. [PubMed] [Google Scholar]
  7. 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]
  8. Lange C. F., Lee R., Merdinger E. Chemistry and end-group analysis on purified M protein of type 12 group A streptococcal cell walls. J Bacteriol. 1969 Dec;100(3):1277–1283. doi: 10.1128/jb.100.3.1277-1283.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. STOLLERMAN G. H., EKSTEDT R. Long chain formation by strains of group A streptococci in the presence of homologous antiserum: a type-specific reaction. J Exp Med. 1957 Sep 1;106(3):345–356. doi: 10.1084/jem.106.3.345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Straus D. C., Lange C. F. Immunochemistry and end-group analyses of group A streptococcal M proteins. Infect Immun. 1972 Jun;5(6):927–932. doi: 10.1128/iai.5.6.927-932.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Vincent W. F., Lisiewski K. J. Improved growth medium for group A streptococci. Appl Microbiol. 1969 Nov;18(5):954–955. doi: 10.1128/am.18.5.954-955.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Wiley G. G., Bruno P. N. Cross-reactions among group A streptococci. II. Further analysis of antigens related to type-specificity and protection. J Immunol. 1969 Aug;103(2):149–154. [PubMed] [Google Scholar]

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