ISOLATION OF MULTIPLE ISOFORMS OF ALPHA-FIBRINOGENASE FROM THE WESTERN DIAMONDBACK RATTLESNAKE, CROTALUS-ATROX - N-TERMINAL SEQUENCE HOMOLOGY WITH ANCROD, AN ANTITHROMBOTIC AGENT FROM MALAYAN VIPER
ISOLATION OF MULTIPLE ISOFORMS OF ALPHA-FIBRINOGENASE FROM THE WESTERN DIAMONDBACK RATTLESNAKE, CROTALUS-ATROX - N-TERMINAL SEQUENCE HOMOLOGY WITH ANCROD, AN ANTITHROMBOTIC AGENT FROM MALAYAN VIPER
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DOI：
10.1006/bbrc.1994.1861
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发表时间：
1994-06-30
影响因子：
3.1
通讯作者：
CHIOU, SH
中科院分区：
文献类型：
--
作者：
HUNG, CC;CHIOU, SH
采用离子交换层析、凝胶渗透层析和反相高效液相色谱等方法从响尾蛇（Crotalus atrox）蛇毒中分离纯化了3种纤溶蛋白酶。通过SDS-凝胶电泳判断，级分显示为均质的。在体外，它们还显示出对纤维蛋白原分子α链的高特异性蛋白水解活性。对这些具有纤维蛋白原酶活性的纯化组分的进一步表征表明，它们都是具有约33，000 - 35，000的相似分子量的单链蛋白。它们在高温下的稳定性进行了检查，并使用氧化的胰岛素B-链作为底物的各种蛋白酶的切割特异性进行了研究。对纤维蛋白原分子的反应性，在不同的蛋白酶抑制剂，包括EDTA，β-巯基乙醇，大豆胰蛋白酶抑制剂，抑肽酶和苯甲磺酰氟的存在下，也进行了研究，以分类这些蛋白酶的反应机制的基础上。氨基酸分析表明，这些纯化的毒液毒素在大多数氨基酸（包括半胱氨酸、甲硫氨酸和酪氨酸）方面具有密切相似的组成。这些蛋白酶的N-末端序列分析表明，它们类似于安克洛，一种抗血栓剂分离的马来蝮蛇。这项研究指出，在响尾蛇科响尾蛇中存在一个新的钩状纤维蛋白原酶家族，这可能是有用的有效的毒液为基础的抗凝剂。(C)1994年学术公司新闻界，
Three fibrinogenolytic proteases from the rattlesnake (Crotalus atrox) venom were isolated and purified from multiple-step chromatographies including ion-exchange chromatography, gel permeation and reversed-phase HPLC. The fractions were shown to be homogeneous as judged by SDS-gel electrophoresis. In vitro, they also showed a high and specific proteolytic activity against alpha-chains of fibrinogen molecules. Further characterization of these purified fractions with fibrinogenase activity indicated that they are all single-chain proteins with similar molecular weights of about 33,000-35,000. Their stability at high temperatures was examined and the cleavage specificity of various proteases was studied using oxidized insulin B-chains as substrates. The reactivity toward fibrinogen molecules in the presence of varied protease inhibitors including EDTA, beta-mercaptoethanol, soybean trypsin inhibitor, aprotinin and phenylmethanesulfonyl fluoride was also investigated in order to classify these proteases based on their reaction mechanisms. Amino acid analysis indicates that these purified venom toxins possess closely-similar compositions with regard to most amino acids including cysteine, methionine and tyrosine. N-Terminal sequence analysis of these proteases revealed that they are similar to ancrod, an antithrombotic agent isolated from the Malayan pit viper. This study points to the existence of a family of novel ancrod-like fibrinogenases in crotalid rattlesnakes, which may be useful as effective venom-based anticoagulants. (C) 1994 Academic Inc. Press,