Cotiarinase is a novel prothrombin activator from the venom of?Bothrops cotiara

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• 作者：Eduardo S. Kitano ; Thalita C. Garcia ; Milene C. Menezes ; Alex ; re K. Tashima ; Andr¨¦ Zelanis ; Solange M.T. Serrano
• 关键词：Bothrops cotiara ; Snake venom ; Snake venom serine proteinase ; Prothrombin activation ; Thrombin
• 刊名：Biochimie
• 出版年：2013
• 出版时间：August, 2013
• 年：2013
• 卷：95
• 期：8
• 页码：1655-1659
• 全文大小：494 K

文摘

Snake venom serine proteinases (SVSPs) may affect hemostatic pathways by specifically activating components involved in coagulation, fibrinolysis and platelet aggregation or by unspecific proteolytic degradation. In this study, we purified and characterized an SVSP from m>Bothrops cotiaram> venom, named cotiarinase, which generated thrombin upon incubation with prothrombin. Cotiarinase was isolated by a two-step procedure including gel-filtration and cation-exchange chromatographies and showed a single protein band with a molecular mass of 29?kDa by SDS-polyacrylamide gel electrophoresis under reducing conditions. Identification of cotiarinase by mass spectrometric analysis revealed peptides that matched sequences of viperid SVSPs. Cotiarinase did not show fibrinogen-clotting, platelet-aggregating, fibrinogenolytic and factor X activating activities. Upon incubation with prothrombin the generation of thrombin was detected using the peptide substrate d-Phe-Pip-Arg-m>pm>NA. Moreover, mass spectrometric identification of prothrombin fragments generated by cotiarinase in the absence of co-factors (phospholipids, factor Va, factor Xa and Ca²⁺ ions), indicated the limited proteolysis of this protein to release prothrombin 1, fragment 1 and thrombin. Cotiarinase is a novel SVSP that acts on prothrombin to release active thrombin that does not match any group of the current classification of snake venom prothrombin activators.