Binding of crotoxin, a presynaptic phospholipase A2 neurotoxin, to negatively charged phospholipid vesicles.
Crotoxin, isolated from the venom of Crotalus durissus terrificus, is a potent neurotoxin consisting of a basic and weakly toxic phospholipase A2 subunit (component B) and an acidic nonenzymatic subunit (component A). The nontoxic component A enhances the toxicity of the phospholipase subunit by preventing its nonspecific adsorption. The binding of crotoxin and of its subunits to small unilamellar phospholipid vesicles was examined under experimental conditions that prevented any phospholipid hydrolysis. Isolated component B rapidly bound with a low affinity (Kapp in the millimolar range) to zwitterionic phospholipid vesicles and with a high affinity (Kapp of less than 1 microM) to negatively charged phospholipid vesicles. On the other hand, the crotoxin complex did not interact with zwitterionic phospholipid vesicles but dissociated in the presence of negatively charged phospholipid vesicles; the noncatalytic component A was released into solution, whereas component B remained tightly bound to lipid vesicles, with apparent affinity constants from 100 to less than 1 microM, according to the chemical composition of the phospholipids. On binding, crotoxin or its component B caused the leakage of a dye entrapped in vesicles of negatively charged but not of zwitterionic phospholipids. The selective binding of crotoxin suggests that negatively charged phospholipids may constitute a component of the acceptor site of crotoxin on the presynaptic plasma membrane.
Laboratoire des Venins, Unité Associée Pasteur/INSERM 285, Institut Pasteur, Paris, France.
F
F
Radvanyi
B
B
Saliou
M P
MP
Lembezat
C
C
Bon
D055598:Chemical Phenomena; D002621:Chemistry; D002850:Chromatography, Gel; D003435:Crotalid Venoms / Q000302:isolation & purification / Q000494:pharmacology; D003439:Crotoxin / Q000302:isolation & purification / Q000494:pharmacology; D007700:Kinetics; D008081:Liposomes; D008968:Molecular Conformation; D009498:Neurotoxins; D010740:Phospholipases / Q000037:antagonists & inhibitors*; D010741:Phospholipases A / Q000037:antagonists & inhibitors*; D054467:Phospholipases A2; D013050:Spectrometry, Fluorescence
D003435:Crotalid Venoms; D008081:Liposomes; D009498:Neurotoxins; D003439:Crotoxin; D010740:Phospholipases; D010741:Phospholipases A; D054467:Phospholipases A2