Isolation and characterization of a thrombin-like enzyme from the venom of Crotalus durissus terrificus

A thrombin-like enzyme was isolated in 6% yield from the venom of Crotalus durissus terrificus by ammonium sulfate precipitation followed by gel filtration on Sephadex G-75 and finally affinity chromatography on Sepharose-1,4-butanediol-diglycyl-p-aminobenzamide eluted with 0.15 M benzamidine. The e...

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Bibliographic Details
Published in:	Brazilian journal of medical and biological research Vol. 19; no. 3; p. 333
Main Authors:	Raw, I, Rocha, M C, Esteves, M I, Kamiguti, A S
Format:	Journal Article
Language:	English
Published:	Brazil 1986
Subjects:	Blood Coagulation Chromatography, Gel Crotalid Venoms - analysis Electrophoresis, Polyacrylamide Gel Molecular Weight Thrombin - isolation & purification
Online Access:	Get more information
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Summary:	A thrombin-like enzyme was isolated in 6% yield from the venom of Crotalus durissus terrificus by ammonium sulfate precipitation followed by gel filtration on Sephadex G-75 and finally affinity chromatography on Sepharose-1,4-butanediol-diglycyl-p-aminobenzamide eluted with 0.15 M benzamidine. The enzyme behaved like a single component on SDS-PAGE corresponding to a molecular weight of 34 kDa. The specific activity of the enzyme toward bovine fibrinogen was 71 NIH U/mg protein. The pH optimum for the coagulation of human fibrinogen was 8.0. The enzyme hydrolyzes the alpha-chain of fibrinogen, has amidase activity on L-arginine-p-nitroanilide and L-arginine-7-amido-4-methyl-coumarin amino terminal blocked peptides and presents esterolytic activity on N-alpha-tosyl-L-arginine-methylester.
ISSN:	0100-879X