In order to dissect out whether multiple activities of cardiotoxins (CTXs) are connected, to some extent, with each other, studies on reduced and S-carboxyamidomethylated (Rcam) Naja naja atra CTX3 were carried out in the present study. Although both CTX3 and Rcam-CTX3 induced apoptotic death of PC-3 cells as evidenced by propodium iodide/annexin V double staining, degradation of procaspases and DNA fragmentation, the cytotoxicity of Rcam-CTX3 was mostly 100-fold lower than that noted with native toxin. However, Rcam-CTX3 retained approximately 38% of the membrane-damaging activity of native toxin as revealed by the decrease in calcein self-quenching from phospholipid vesicles. These results are likely to reflect that the mechanism of cytotoxicity by CTX3 is not heavily dependent on its membrane-perturbing effect, and suggest that the structural elements within CTX3 responsible for the two activities are probably separated.
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