Rapid purification of serine proteinases from Bothrops alternatus and Bothrops moojeni venoms

Abstract

Envenomation by Bothrops species results, among other symptoms, in hemostatic disturbances. These changes can be ascribed to the presence of enzymes, primarily serine proteinases some of which are structurally similar to thrombin and specifically cleave fibrinogen releasing fibrinopeptides. A rapid, three-step, chromatographic procedure was developed to routinely purify serine proteinases from the venoms of Bothrops alternatus and Bothrops moojeni. The serine proteinase from B. alternatus displays an apparent molecular mass of ~32 kDa whereas the two closely related serine proteinases from B. moojeni display apparent molecular masses of ~32 kDa and ~35 kDa in SDS-PAGE gels. The partial sequences indicated that these enzymes share high identity with serine proteinases from the venoms of other Bothrops species. These proteins coagulate plasma and possess fibrinogenolytic activity but lack fibrinolytic activity.

Keywords: Bothrops alternatus; Bothrops moojeni; Crude venom; Fibrinogenolysis; MCD; PAGE; Proteolytic activity; SDS; SVSPs; Serine proteinases; minimum coagulant dose; polyacrylamide gel electrophoresis; snake venom serine proteinases; sodium dodecyl sulfate.