• Biomolecules & Therapeutics
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• 제3권2호
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• pp.159-164
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• 1995

Fibrinolytic and fibrinogenolytic activities of the venoms from the Korean snakes, Agkistrodon caliginosus, nosus, Agkistrodon saxatilis and Agkistrodon blomhoffi brevicaudus were compared by fibrin-plate method and polyacrylamide gel electrophoresis, respectively. The venom from A. blomhoffi brevicaudus showed the highest degree of fibrin(ogen)olytic activity, and a protease with the fibrin(open)olytic activity was purified by p-amino-benzamidine affinity chromatography and DEAE ion-exchange chromatography. The purified enzyme had a molecular weight of 50,800 and a capability to degrade the B$\beta$-chain of fibrinogen preferentially to the $A\alpha$-chain, but not the ${\gamma}$-chain. Fibrinolytic activity of the purified enzyme was approximately 3.8 plasmin unit/mg protein.

• Biomolecules & Therapeutics
• /
• 제3권2호
• /
• pp.165-170
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• 1995

The biochemical properties of the fibrinolytic protease of 50,800 Da isolated from the venom of Kgdistrodon blomhoffi brevicaudus were characterized. The enzyme hydrolyzed the carboxyl side of arginine in the synthetic chromogenic peptides, N-Benzoyl-Phe-Val-Arg-pNA and N-p-Tosyl-Gly-Pro-Arg-pNA, and the enzyme activity was inhibited by phenylmethylsulfonylfluoride indicating that the enzyme belongs to the serine protease family. The pretense showed maximum activity at pH 7.5 and inhibited by ZnCl$_2$, CuSO$_4$, but not by soybean trypsin inhibitor, pepstatin A, 2-mercaptoethanol and EDTA. The fm value determined with N-p-Tosyl-Gly-Pro-Arg-pNA was 0.2 mM. The thrombolytic activity of the purified enzyme was evaluated by platelet aggregation test in rabbits. While the platelet count ratio in blood of the rabbits injected with thrombin alone declined from 1.0 to 0.6 within 7 min and maintained around 0.6 for 24 hours thereafter, the ratio rapidly recovered from around 0.6 to 0.8 in 1 hr, to 1.0 in 24 hrs when the rabbits were sequentially treated with thrombin and the purified enzyme. The result showed that the serine protease from A. blomhoffi brevicoudus of 50,800 Da had a thrombolytic activity in vivo and the enzyme might be developed as a therapuetic agent for the treatment of thrombic disease.