蛇毒の酵素化学的研究(第3報)ヒャッポダ毒より血液凝固作用を有するエステラーゼの精製およびその性質

概要

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A human blood coagulation principle was isolated from the venom of a snake Agkistrodon acutus, by a combination of gel filtration on Sephadex G-75 and chromatographies on DEAE-Sephadex A-50,CM-Sephadex C-50,and DEAE-Sephadex A-50. By these procedures, 17.4 mg purified preparation was obtained from 1 g of crude venom. This coagulation principle hydrolyzed arginine esters, such as tosyl-l-arginine methyl ester (TAME) or benzoyl-arginine ethyl ester (BAEE), but did not digest casein. The coagulation and esterolytic activities were inhibited by diisopropyl fluorophosphate (DEP) or antiserum but not by soy bean trypsin inhibitor (SBTI), ethylene diamine tetra acetic acid (EDTA), cysteine, heparin, or p-chloromercuribenzoate (PCMB). The preparation was homogeneous as judged by disc electrophoresis on polyacrylamide gel and isoelectric focusing. The molecular weight of this protein was determined to be approximately 52000,and the isoelectric point was found to be pH 4.7 by isoelectric focusing with carrier ampholyte. The coagulation and esterolytic activities of this protein were 37.5 and 99.3 unit, respectively. This protein was stable to heat treatment, and between pH 6 and 8. Michaelis constant (Km) and inhibition constant (K_1) for this protein were found to be 1.34×10^<-3> M and 1.92×10^<-3> M, respectively. This enzyme had some carbohydrates but did not contain any nucleic acids.
公益社団法人日本薬学会の論文
1978-07-25