Ouyang C, Hwang L J, Huang T F
Toxicon. 1983;21(1):25-33. doi: 10.1016/0041-0101(83)90046-6.
By means of DEAE-Sephadex A-50 column chromatography, Agkistrodon rhodostoma (Malayan pit viper) snake venom was separated into eleven fractions. Fraction II had fibrinogenolytic activity, and when further purified by gel filtration was homogeneous, as judged by sodium dodecylsulfate polyacrylamide gel electrophoresis. It had a single peptide chain with a molecular weight of 25,360 and an isoelectric point greater than 10. The fibrinogenolytic activity was completely destroyed after heating for 30 min at 60 degrees C at pH 5.6, 7.4 or 8.8. This enzyme cleaved specifically the alpha(A) chain of monomeric fibrinogen, without cleaving the beta(B) chain or gamma chain. The specific fibrinogenolytic activity was 51 mg fibrinogen/min per mg protein. This enzyme showed proteolytic activities toward fibrinogen, fibrin and casein, but was devoid of phospholipase A and tosyl-L-arginine methylester esterase activities which are found in the crude venom. The fibrinogenolytic activity was inhibited by EDTA and cysteine, but not by epsilon-aminocaproic acid.
通过DEAE-葡聚糖A-50柱色谱法,将红口蝮蛇(马来亚蝮蛇)蛇毒分离成11个组分。组分II具有纤维蛋白原溶解活性,通过凝胶过滤进一步纯化后,经十二烷基硫酸钠聚丙烯酰胺凝胶电泳判断为均一。它有一条单肽链,分子量为25,360,等电点大于10。在pH 5.6、7.4或8.8条件下于60℃加热30分钟后,纤维蛋白原溶解活性完全丧失。该酶特异性切割单体纤维蛋白原的α(A)链,而不切割β(B)链或γ链。比纤维蛋白原溶解活性为每毫克蛋白质每分钟51毫克纤维蛋白原。该酶对纤维蛋白原、纤维蛋白和酪蛋白表现出蛋白水解活性,但不具有粗毒液中存在的磷脂酶A和甲苯磺酰-L-精氨酸甲酯酯酶活性。纤维蛋白原溶解活性受EDTA和半胱氨酸抑制,但不受ε-氨基己酸抑制。