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Purification and cloning of cysteine-rich proteins from Trimeresurus jerdonii and Naja atra venoms

机译:细叶Trimeresurus jerdonii和眼镜蛇眼镜蛇毒液中富含半胱氨酸的蛋白的纯化和克隆

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摘要

Three 26 kDa proteins, named as TJ-CRVP, NA-CRVP1 and NA-CRVP2, were isolated from the venoms of Trimeresurus jerdonii and Naja atra, respectively. The N-terminal sequences of TJ-CRVP and NA-CRVPs were determined. These components were devoid of the enzymatic activities tested, such as phospholipase A(2), arginine esterase, proteolysis, L-amino acid oxidase, 5' nucleotidase, acetylcholinesterase. Furthermore, these three components did not have the following biological activities: coagulant and anticoagulant activities, lethal activity, myotoxicity, hemorrhagic activity, platelet aggregation and platelet aggregation-inhibiting activities. These proteins are named as cysteine-rich venom protein (CRVP) because their sequences showed high level of similarity with mammalian cysteine-rich secretory protein (CRISP) family. Recently, some CRISP-like proteins were also isolated from several different snake venoms, including Agkistrodon blomhoffi, Trimeresurus flavoviridis, Lanticauda semifascita and king cobra. We presumed that CRVP might be a common component in snake venoms. Of particular interest, phylogenetic analysis and sequence alignment showed that NA-CRVP1 and ophanin, both from elapid snakes, share higher similarity with CRVPs from Viperidae snakes.
机译:从Jerdonii的毒液和Naja atra的毒液中分别分离出三个26 kDa的蛋白,分别命名为TJ-CRVP,NA-CRVP1和NA-CRVP2。确定了TJ-CRVP和NA-CRVP的N末端序列。这些成分没有测试的酶活性,如磷脂酶A(2),精氨酸酯酶,蛋白水解,L-氨基酸氧化酶,5'核苷酸酶,乙酰胆碱酯酶。此外,这三个成分不具有以下生物学活性:凝结和抗凝活性,致死活性,肌毒性,出血活性,血小板聚集和血小板聚集抑制活性。这些蛋白质被称为富含半胱氨酸的毒液蛋白(CRVP),因为它们的序列与哺乳动物富含半胱氨酸的分泌蛋白(CRISP)家族表现出高度的相似性。最近,还从几种不同的蛇毒中分离出了一些类似于CRISP的蛋白质,包括蛇毒(Agkistrodon blomhoffi),Trimeresurus flavoviridis,Lanticauda semifascita和眼镜王蛇。我们认为CRVP可能是蛇毒中的常见成分。系统发育分析和序列比对特别有趣,是来自于蛇类的NA-CRVP1和ophanin与来自蛇形蛇的CRVP具有更高的相似性。

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