DISPHOLIDUS-TYPUS (BOOMSLANG) SNAKE-VENOM - PURIFICATION AND PROPERTIES OF THE COAGULANT PRINCIPLE

Peter C. Hiestand and Rosemarie R. Hiestand, Dispholidus typus (Boomslang) snake venom: purification and properties of the coagulant principle. Toxicon 17, 489-498, 1979.-Dispholidus typus venom was fractionated into five fractions by means of CM-cellulose column chromatography. Fraction 1 exhibited strong coagulant activity on citrated rabbit plasma. This fraction was further purified by means of preparative isoelectric focusing, DEAE-cellulose chromatography, and gel filtration on Sephadex G-200. After gel filtration two fractions (fraction c and d) containing coagulant activity were obtained. Fraction d separated on disc gel electrophoresis at pH 8·3 into six bands of which one (d1) coincided with fraction c upon isoelectric focusing and SDS* * SDS = Sodium dodecyl sulfate. gel electrophoresis. Fraction c which contains the main coagulant activity in Dispholidus typus venom was homogeneous on disc gel electrophoresis and on isoelectric focusing. The molecular weight estimated by gel filtration on Sephadex G-200 was 55,000 and by electrophoresis in sodium dodecyl sulfate 67,000. The isoelectric point was pH 4·4. Chemical analysis showed that fraction c was a thermolabile glycoprotein. Its coagulant activity was approximately 10 times higher than that of crude venom since the active fractions account for ca. 10% of material found in crude dry venom. Although it did exhibit most of the coagulant activity, fraction c did not possess any of the caseinolytic or BAEE† † BAEE = N-benzoyl-arginine ethyl ester.-hydrolyzing activities found in crude venom. © 1979.