Structural determinant of the vesicle aggregation activity of annexin I
Structural determinant of the vesicle aggregation activity of annexin I
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DOI：
10.1021/bi990457p
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发表时间：
1999-10-19
期刊：
影响因子：
2.9
通讯作者：
Cho, W
中科院分区：
文献类型：
--
作者：
Bitto, E;Cho, W
一些膜联蛋白，包括膜联蛋白I、II、IV和VII，可以促进膜聚集。为了鉴定参与膜联蛋白I介导的膜聚集的氨基酸，我们产生了缺乏氨基末端各个部分的人膜联蛋白I的截短突变体。这些突变体的体外囊泡结合和聚集活性表明膜联蛋白I的氨基末端区域跨越残基26-29和羧基末端核心都参与膜聚集。由膜联蛋白I的残基24-35和膜联蛋白V的核心组成的嵌合体具有显著高于膜联蛋白V但低于膜联蛋白I的囊泡聚集活性的发现进一步支持了这一观点。进一步的位点特异性突变的氨基末端区域的膜联蛋白I表明，赖氨酸-26和赖氨酸-29是必不可少的膜聚集活性。游离和囊泡结合的野生型和K29 E突变体的胰蛋白酶消化模式的比较表明，赖氨酸-26和29赖氨酸的主要作用是诱导和稳定膜联蛋白I的活性构象囊泡聚集。
Some annexins, including annexins I, II, TV, and VII, can promote membrane aggregation. To identify amino acids involved in annexin I-mediated membrane aggregation, we generated truncated mutants of human annexin I lacking Various parts of the amino terminus. The in vitro vesicle binding and aggregation activities of these mutants indicated that both the amino-terminal region of annexin I spanning residues 26-29 and the carboxy-terminal core are involved in membrane aggregation. This notion was further supported by the finding that a chimera composed of residues 24-35 of annexin I and the core of annexin V has vesicle aggregation activity that is significantly higher than that of annexin V but lower than that of annexin I. Further site-specific mutations in the amino-terminal region of annexin I indicated that Lys-26 and Lys-29 are essential for its membrane aggregation activity. The comparison of tryptic digest patterns of free and vesicle-bound wild type and K29E mutant suggests that a primary role of Lys-26 and Lys-29 is to induce and stabilize an active conformation of annexin I for vesicle aggregation.