Basic research on development of anti-thrombus drugs that mimic disintegrins

模拟解整合素抗血栓药物开发的基础研究

基本信息

批准号：
07558218
负责人：
KAWABATA Shun-ichiro
金额：
$ 1.15万
依托单位：
KYUSHU UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07558218/
关键词：
Disintegrin snake venom Thrombosis

项目摘要

The complete amino acid sequence of ecarin is deduced from the nucleotide sequence of a cDNA clone isolated by screening a venomous gland cDNA library of Kenyan Echis (E).carinatus. The cDNA sequence with 2,379 base pairs encodes an open reading frame of 616 amino acids with a remakeable sequence homology to the putative precursor protein of trigramin from Trimeresurus gramineus venom (61% identity) and a large hemorrhagin, jararhagin, from the pit viper Bothrops jararaca venom (62% identity). Thus, ecarin, as well as jararhagin and trigramin, is translated as a precursor protein, which may be processed posttranslationally. The ecarin proprotein has a "cysteine switch" motif (-Pro-Lys-Met-Cys-Gly-Val-) similar to that involved in the activation of matrix metalloptoteinase zymogens. The processed mature protein consists of 426 amino acid residues (residues 191-616), showing the strongest sequence similarity with that of Russell's viper venom factor X activator (RVV-X) heavy chain (64% identity). Like RVV-X heavy chain, ecarin contains metalloproteinase, disintegrin and cysteine-rich domains. The metalloproteinase domain has a typical zinc chelating sequence (-His-Glu-Xaa-Xaa-His-Xaa-Xaa-Gly-Xaa-Xaa-His-) found in crayfish astacin. In the disintegrin domain of ecarin, the Arg-Gly-Asp sequence is replaced by Arg-Asp-Asp, as found in the disintegrin domains of RVV-X heavy chain (Arg-Asp-Glu) and a guinea pig sperm fusion protein PH-30b (Thr-Asp-Glu). These findings show that while there are the structual and evolutionary relationships among these proteins, each has diverse functional activity.

ecarin 的完整氨基酸序列是从通过筛选肯尼亚 Echis (E).carinatus 毒腺 cDNA 文库分离的 cDNA 克隆的核苷酸序列推导出来的。具有 2,379 个碱基对的 cDNA 序列编码一个 616 个氨基酸的开放阅读框，与来自竹叶青毒液的三谷蛋白的推定前体蛋白（61% 同一性）和来自坑蝰蛇 Bothrops jararaca 的大出血素 jararhagin 具有可重构的序列同源性毒液（62% 同一性）。因此，ecarin以及jararhagin和trigramin被翻译为前体蛋白，可以在翻译后进行加工。 ecarin 蛋白原具有一个“半胱氨酸开关”基序 (-Pro-Lys-Met-Cys-Gly-Val-)，类似于参与基质金属蛋白酶酶原激活的基序。加工后的成熟蛋白由 426 个氨基酸残基（残基 191-616）组成，与拉塞尔毒蛇毒 X 因子激活剂（RVV-X）重链的序列相似性最强（64% 同一性）。与 RVV-X 重链一样，ecarin 含有金属蛋白酶、解整合素和富含半胱氨酸的结构域。金属蛋白酶结构域具有小龙虾虾红素中发现的典型锌螯合序列（-His-Glu-Xaa-Xaa-His-Xaa-Xaa-Gly-Xaa-Xaa-His-）。在 ecarin 的解整合素结构域中，Arg-Gly-Asp 序列被 Arg-Asp-Asp 取代，如在 RVV-X 重链 (Arg-Asp-Glu) 的解整合素结构域和豚鼠精子融合蛋白 PH 中发现的那样-30b（苏氨酸-天冬氨酸-谷氨酸）。这些发现表明，虽然这些蛋白质之间存在结构和进化关系，但每种蛋白质都具有不同的功能活性。