Natural phosphorylation of group A streptococcal pyrogenic exotoxin type C.

Group A streptococcal pyrogenic exotoxin (SPE) type C, produced by strain T18P grown in the presence of 32P, was separated from culture supernatant fluids by using alcohol precipitation. The resulting toxin (EtOH-1) contained 3 X 10(6) to 5 X 10(6) cpm of 32P per milligram of protein. The radiolabel migrated with SPE C during isoelectric focusing in polyacrylamide gels (pI 6.7) and double immunodiffusion, in which the toxin formed a line of identity with highly purified SPE C when reacted with hyperimmune antisera raised against SPE C. The EtOH-1 radiolabeled toxin was pyrogenic and had the capacity to enhance host susceptibility to lethal endotoxin shock. EtOH-1 toxin lost both radiolabel and biological activity after being treated with alkaline phosphatase. The nonspecific lymphocyte mitogenicity of purified unlabeled SPE C was stimulated by adenosine monophosphate but not adenosine, adenosine diphosphate, or adenosine triphosphate. Adenosine monophosphate may function as a cofactor of SPE C and contribute the phosphate group required for biological activity.