An activator of blood coagulation factor X from the venom of Bungarus fasciatus.
A specific activator of blood coagulation factor X was purified from the venom of Bungarus fasciatus by gel filtration and by ion-exchange chromatography on a Mono-Q column (FPLC). It consisted of a single polypeptide chain, with a mol. wt of 70,000 in reducing and non-reducing conditions. The enzyme had an amidolytic activity towards the chromogenic substrates S-2266 and S-2302 but it did not hydrolyse S-2238, S2251 or S-2222, which are specific substrates for thrombin, plasmin and factor Xa, respectively. The enzyme activated factor X in vitro and the effect was Ca2+ dependent with a Hill coefficient of 7.9. As with physiological activators, the venom activator cleaves the heavy chain of factor X, producing the activated factor Xa alpha. The purified factor X activator from B. fasciatus venom did not activate prothrombin, nor did it cleave or clot purified fibrinogen. The amidolytic activity and the factor X activation activity of the factor X activator from B. fasciatus venom were readily inhibited by serine protease inhibitors such as diisopropyl fluorophosphate (DFP), phenylmethanesulfonyl fluoride (PMSF), benzamidine and by soybean trypsin inhibitor but not by EDTA. These observations suggest that the factor X activator from B. fasciatus venom is a serine protease. It therefore differs from those of activators obtained from Vipera russelli and Bothrops atrox venoms, which are metalloproteinases.
Zhang
Y
Y
Unité des Venins, Institut Pasteur, Paris, France.
Xiong
Y L
YL
Bon
C
C
D000818:Animals; D001550:Benzamidines / Q000494:pharmacology; D002038:Bungarotoxins / Q000737:chemistry*; D017838:Bungarus; D002118:Calcium / Q000378:metabolism; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D005170:Factor X / Q000187:drug effects*; D005340:Fibrinogen / Q000378:metabolism; D008970:Molecular Weight; D011516:Prothrombin / Q000378:metabolism; D012697:Serine Endopeptidases / Q000302:isolation & purification* / Q000378:metabolism / Q000494:pharmacology; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology; D013379:Substrate Specificity; D014361:Trypsin Inhibitors / Q000494:pharmacology
D001550:Benzamidines; D002038:Bungarotoxins; D015842:Serine Proteinase Inhibitors; D014361:Trypsin Inhibitors; D011516:Prothrombin; D005170:Factor X; D005340:Fibrinogen; D012697:Serine Endopeptidases; C032157:benzamidine; D002118:Calcium