STUDIES ON EPSILON-PROTOTOXIN OF CLOSTRIDIUM PERFRINGENS TYPE D .I. PRUIFICATION METHODS - EVIDENCE FOR MULTIPLE FORMS OF EPSILON-PROTOTOXIN

Clostridium perfringes type D was grown in protein-free medium. ε{lunate}-Prototoxin was found to contain insignificant amounts of ε{lunate}-toxin. Purification of ε{lunate}-prototoxin by column chromatography on DEAE-cellulose followed by CM-cellulose revealed multiple forms of ε{lunate}-prototoxin which differed in their electrophoretic mobility but were immunochemically indistinguishable. The multiple forms of ε{lunate}-prototoxin varied in their activation ratios and specific activities. The most potent preparation contained 3 × 106 to 4.2 × 106 MLD/mg protein which was formed of three electrophoretically different yet immunochemically indistinguishable bands. Activation of ε{lunate}-prototoxin to toxin by treatment with trypsin resulted in a more negatively charged molecule, suggesting splitting of a basic peptide (s). The ε{lunate}-prototoxin and ε{lunate}-toxin show complete immunologic identity. © 1969.