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首页> 外文期刊>Journal of Pharmacopuncture >Characterization of Fibrinolytic Proteases from Gloydius blomhoffii siniticus Venom
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Characterization of Fibrinolytic Proteases from Gloydius blomhoffii siniticus Venom

机译：中华绒螯蟹毒液纤溶蛋白酶的表征

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Objectives : This study was undertaken to identify fibrinolytic proteases from Gloydius blomhoffii siniticus venom and to characterize a major fibrinolytic protease purified from the venom. Methods : The venom was subjected to chromatography using columns of Q-Sepharose and Sephadex G-75. The molecular weights of fibrinolytic proteases showing fibrinolytic zone in fibrin plate assay were determined in SDS-PAGE (Sodium dodecyl sulfate-polyacrylamide gel electrophoresis) The effects of inhibitors and metal ions on fibrinolytic protease and the proteolysis patterns of fibrinogen, gelatin, and bovine serum albumin were investigated. Results : 1) The fibrinolytic fractions of the three peaks isolated from Gloydius blomhoffii siniticus venom contained two polypeptides of 46 and 59 kDa and three polypeptides of 32, 18, and 15 kDa and a major polypeptide of 54 kDa, respectively. 2) The fibrinolytic activity of the purified protease of 54 kDA was inhibited by metal chelators, such as EDTA, EGTA, and 1,10-phenanthroline, and disulfhydryl-reducing compounds, such as dithiothreitol and cysteine. 3) Calcium chloride promoted the fibrinolytic activity of the protease, but mercuric chloride and cobalt(II) chloride inhibited it. 4) The fibrinolytic protease cleaved preferentially A -chain and slowly B -chain of fibrinogen. It also hydrolyzed gelatin but not bovine serum albumin. Conclusions : The Gloydius blomhoffii siniticus venom contained more than three fibrinolytic proteases. The major fibrinolytic protease was a metalloprotease which hydrolyzed both fibrinogen and gelatin, but not bovine serum albumin.
机译：目的：进行这项研究以鉴定中华绒螯蟹毒液中的纤溶蛋白酶，并鉴定从毒液中纯化出的主要纤溶蛋白酶。方法：使用Q-Sepharose和Sephadex G-75色谱柱对蛇毒进行色谱分离。在SDS-PAGE（十二烷基硫酸钠-聚丙烯酰胺凝胶电泳）中测定在纤维蛋白平板测定中显示纤维蛋白溶解区的纤维蛋白溶解蛋白酶的分子量。抑制剂和金属离子对纤维蛋白水解蛋白酶的影响以及纤维蛋白原，明胶和牛血清的蛋白水解模式研究白蛋白。结果：1）从中华草蛇毒中分离出的三个峰的纤溶部分分别含有两个46和59kDa的多肽和三个32、18和15kDa的多肽和一个主要的54kDa的多肽。 2）纯化的54 kDA蛋白酶的纤溶活性被金属螯合剂（如EDTA，EGTA和1,10-菲咯啉）和二巯基还原性化合物（如二硫苏糖醇和半胱氨酸）抑制。 3）氯化钙促进了蛋白酶的纤溶活性，但是氯化汞和氯化钴（II）抑制了它的水解。 4）纤维蛋白水解蛋白酶优先切割血纤蛋白原的A链和缓慢的B链。它也水解明胶，但不水解牛血清白蛋白。结论：中华姬蛇毒中含有三种以上的纤溶蛋白酶。主要的纤维蛋白水解蛋白酶是金属蛋白酶，其水解纤维蛋白原和明胶，但不水解牛血清白蛋白。

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