Peptic digestion of streptococcal M protein. II. Extraction of M antigen from group A streptococci with pepsin.
M protein of group A streptococci was extracted by mild peptic digestion. Optimal amounts of type-specific M protein were released after 20 min of digestion with 0.02 mg of pepsin per ml at pH 5.8. Immunological analysis revealed that, unlike conventional HCl extracts, pepsin extracts lacked the surface C carbohydrate antigen and contained less non-type-specific, heat-stable cellular antigens; they also lacked detectable heat-labile T protein. Similar to HCl extracts, however, the pepsin-extracted M protein precipitated homologous-type M antisera and inhibited type-specific opsonization of homologous group A streptococci. Furthermore, the pepsin extract was capable of inducing type-specific opsonic M antibody in rabbits. This method may provide a useful initial step in the purification of M protein by reducing contaminating antigens.
D000372:Agglutination Tests; D000818:Animals; D000918:Antibody Specificity; D000942:Antigens, Bacterial / Q000302:isolation & purification*; D003168:Complement Fixation Tests; D000939:Epitopes; D006386:Hemagglutination Tests; D007106:Immune Sera; D007114:Immunization; D005779:Immunodiffusion; D008722:Methods; D009895:Opsonin Proteins / Q000032:analysis; D010434:Pepsin A / Q000494:pharmacology*; D011817:Rabbits / Q000276:immunology; D013297:Streptococcus pyogenes / Q000276:immunology*
D000942:Antigens, Bacterial; D000939:Epitopes; D007106:Immune Sera; D009895:Opsonin Proteins; D010434:Pepsin A