The mechanism of inhibition of phospholipase activity of crotoxin B by crotoxin A.

In the crotoxin complex isolated from Crotalus durissus terrificus venom, the component A inhibits the phospholipase A2 activity of crotoxin B only when the substrate is in the aggregated form, preventing the interaction of the enzyme with lecithin--water interfaces. In contrast, with similar rates of hydrolysis of dihexanoyllecithin monomers, the activity of the crotoxin complex is lower than that of crotoxin B when the substrate is aggregated into micelles. Crotoxin B readily hydrolyses dimyristoyllecithin vesicles, the rate being modulated by the physical state of the phospholipid, suggesting that the enzyme is tightly bound to the interface. With the crotoxin complex the rate of vesicle hydrolysis is much slower (about 1/10 that of crotoxin B) and is little affected by the physical state of the lecithin. Direct binding experiments demonstrate that, in contrast to crotoxin B, the crotoxin complex is unable to interact with lecithin--water interfaces. Together with the free accessibility of the enzyme active site in the crotoxin complex, this evidence suggests that a specific area on the enzyme surface, different from the active site and shielded by crotoxin A in the complex, is responsible for the interaction of crotoxin B with lipid--water interfaces.