Extraction of streptococcal type 12 M protein by cyanogen bromide.
Conditions for the release of streptococcal type 12 M protein from whole cells by cyanogen bromide are described; they demonstrated that methionine is not essential to the structural arrangements which account for some of its immunological and biological properties. The released M protein was separated from other proteins by column chromatography with hydroxylapatite. The type-specific molecules which reacted with precipitating antibodies were found only in the 0.3 M eluate, formed zones with mobilities less than 12% of that of the dye front on electrophoresis in the standard acrylamide disc gel system, formed at least four bands in sodium dodecyl sulfate-acrylamide disc gels with molecular weights ranging from 12,000 to 23,000, and stimulated the formation of opsonic antibodies in rabbits. Cyanogen bromide provides a highly specific method for the release of M proteins which should prove particularly useful in analyses of structural-functional relationships among different M proteins.
D000596:Amino Acids; D000818:Animals; D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001426:Bacterial Proteins / Q000302:isolation & purification*; D002845:Chromatography; D003488:Cyanogen Bromide / Q000494:pharmacology*; D004591:Electrophoresis, Polyacrylamide Gel; D005612:Freeze Drying; D006882:Hydroxyapatites; D009895:Opsonin Proteins; D011817:Rabbits; D013297:Streptococcus pyogenes
D000596:Amino Acids; D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D006882:Hydroxyapatites; D009895:Opsonin Proteins; D003488:Cyanogen Bromide