NEW METHOD FOR PURIFICATION OF THE THROMBIN-LIKE ENZYME FROM THE VENOM OF THE EASTERN DIAMONDBACK RATTLESNAKE

Crotalase, a thrombin-like enzyme from the venom of eastern diamondback rattlesnake (Crotalus adamanteus), has been purified by a new procedure consisting of gel filtration on Sephadex G-100, DEAE-cellulose chromatography, benzamidine-Sepharose affinity chromatography and SP-Sephadex chromatography. Microheterogeneity was observed after either acrylamide gel electrophoresis or isoelectric focusing. Neuraminidase treatment showed the microheterogeneity to be due to varying levels of sialic acid. The isoelectric point of the desialylated enzyme is 4.6. Coagulant activity was not diminished by neuraminidase treatment. However, there was a slight increase in amidolytic activity on a synthetic chromogenic substrate (S-2238). © 1979.