Skip to main content
Applied Microbiology logoLink to Applied Microbiology
. 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.

Full text

PDF
28

Images in this article

Selected References

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

  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]

Articles from Applied Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES