A mechanism for regulation of clostripain activity and modification of inflammatory response by the enzyme.

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To examine a pathogenic role of clostripain(Clp) produced by Clostridium perfringens, Clp mutant and wild-type strains were injected into the mouse femur. Both strains showed gas gangrene and leukostasis to a similar extent, but no significant difference in histological characteristics. We developed an in-frame deletion method, by which we constructed various mutant strains. To obtain various toxins and mutant enzymes in large quantities, we constructed xylose-inducible expression system. Although we attempted to analyze regulation of Clp activity by a linker peptide and also an effect of Clp on eat-me signal of polymorphonuclear leukocytes, we failed to obtain conclusive evidence. We found the Clp mutant aggregates upon heating. Addition of EDTA, an inhibitor of Clp, to the culture of the wild type strain caused self-aggregation.