Fractionation and partial characterization of toxic components of timber rattlesnake venom

Abstract The venom of the timber rattlesnake, Crotalus horridus horridus , has been separated into five distinct fractions using diethylaminoethyl cellulose ion-exchange chromatography with elution by an increasing sodium acetate gradient and pH adjustment. Three of these five fractions are lethal and hemorrhage-inducing in mice and also have the ability to catalyze the hydrolysis of hide powder. Of these three, one (fraction I a ) is not retained by the diethylaminoethyl cellulose. This fraction has been further separated into five fractions (A, B, C, D and E) by ion-exchange chromatography on carboxymethyl cellulose. Two distinct lethal activities are apparent in these five fractions. The most basic of these (fraction E) appears to contain only a single protein component of mol. wt 17,000 ± 1000 as determined by SDS gel electrophoresis. In mice it is lethal with paralysis of limbs and respiratory distress apparent prior to death. It has none of the hemorrhagic or enzymatic activities of the whole venom. The other lethal activity is present mainly in fraction B. It consists of, at least, seven protein components and contains the phosphodiesterase and hemorrhagic activities of fraction I a . It also contains a very weak hide powder hydrolyzing activity. The location of a number of previously reported activities among the various fractions obtained from the two types of chromatographies has been determined.