大蹼铃蟾皮肤膜联蛋白Ⅱ相关蛋白的分离纯化及活性

Purification and Characterization of Annexin ⅡRelated Protein from Bombina maxina Skin

通过阴离子交换、凝胶过滤和阳离子交换层析,从大蹼铃蟾皮肤中纯化到一个表观分子量为33kDa的单链蛋白。N-末端序列比较分析显示,该蛋白与来自非洲爪蟾、红色原鸡和人膜联蛋白Ⅱ的N-末端序列相同的氨基酸分别占70%、64%和56%。该蛋白具有以钙依赖的方式抑制专一性血小板膜糖蛋白Ⅵ受体激动剂———Stejnulxin诱导洗涤人血小板聚集的生物学功能,最大抑制率达48%。结合其N-末端序列BLAST搜索结果及其活性的钙依赖性,推测该蛋白是与膜联蛋白Ⅱ相关的一类蛋白质。

A single chain protein with an apparent molecular weight of 33 kDa was purified from skin of Bombina maxima by a combination of ion exchange and gel filtration chromatography steps. N-terminal amino acid sequence determination indicated that it shares 70%, 64% and 56% identity with those of annexin Ⅱ from the African claw toad, red jungle fowl and human, respectively. The purified protein from B. maxina inhibits stejnulxin （a specific platelet agonist via platelet membrane glycoprotein Ⅵ receptor） induced platelet aggregation in a Ca^2＋ dependent manner. Maximal inhibition rate reaches 48%. Based on the N-terminal amino acid sequence BLAST search results and the fact that its activity is strictly Ca^2＋ dependent, the purified protein might be structurally and functionally related to the annexin protein family.