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的有毒腺cDNA文库（E）.arinatus分离的cDNA克隆的核苷酸序列推导的。具有2,379个碱基对的cDNA序列编码了616个氨基酸的开放式阅读框，具有可重复的序列同源性，该序列同源于提取的三毛素的前体蛋白质蛋白（trimeresurus gramineus venom（61％身份）（61％身份））和大型出血，jararhagin，jararhagin，jararhagin，jararhagin，jararhagin，pitiperpitip viperops jararaca venom（62％）（62％）。因此，Ecarin，以及Jararhagin和Trigramin被翻译为前体蛋白，可以在翻译后进行处理。 Ecarin Proprotein具有“半胱氨酸开关”基序（-pro-lys-met-cys-gly-val-），类似于与基质金属蛋白酶Zymogens的激活相似。加工后的成熟蛋白质由426个氨基酸残基（残基191-616）组成，显示与罗素的Viper毒素毒性因子X激活剂（RVV-X）重链（64％身份）的序列相似性最强。像RVV-X重链一样，Ecarin含有金属蛋白酶，崩解蛋白和富含半胱氨酸的域。金属蛋白酶结构域具有典型的锌螯合序列（-his-glu-xaa-xaa-xaa-xaa-xaa-xaa-xaa-xaa-gly-xaa-xaa-xaa-his-）。在ECARIN的解体蛋白结构域中，如RVV-X重链（ARG-ASP-GLU）的解体蛋白结构域（ARG-ASP-GLU）和豚鼠精子融合蛋白PH-30B（THR-ASP-GLU）所发现的那样，Arg-Gly-Asp序列被Arg-Asp-Asp取代。这些发现表明，尽管这些蛋白质之间存在结构和进化关系，但每个蛋白质都具有多种功能活性。