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东亚钳蝎一种新的具有心脏毒性的α-样毒素的纯化、结晶及初步结构解析

Purification, crystallization and initial structural solution of a new alpha-like toxin with cardiac toxicity from scorpion Buthus martensii Karsch.

作者信息

Guan Rong-Jin, He Xiao-Lin, Wang Miao, Li Gen-Pei, Wang Da-Cheng

机构信息

Center for Molecular Biology, Institute of Biophysics, Chinese Academy of Sciences, 100101 Beijing China.

出版信息

Protein Pept Lett. 2002 Oct;9(5):441-9. doi: 10.2174/0929866023408562.

Abstract

An alpha-like toxin named BmK M7 active on both mammals and insects has been purified from the venom of scorpion Buthus martensii Karsch (BmK) recently. The electrophysiological experiments showed that M7 can bind to human cardiac Na+-channel and modify its normal properties, hence can be considered as a cardiotoxin. Single crystals of M7 have been obtained by hanging-drop vapor diffusion method using ammonium sulfate as precipitant in Tris-HCl buffer at pH 8.5. A data set to 1.40 A resolution was collected using synchrotron radiation and CCD detector in Photon Factory in Japan. Data analysis showed that the crystals belonged to space group P3(1)21/P3(1)21, with cell dimensions a=b=32.76 A, c=176.82 A. Assuming two molecules per asymmetric unit, the Vm value is 1.92 A3/Da. The initial structural analysis was carried out by molecular replacement, which showed the correct space group (P3(1)21), and the orientations and positions of the two molecules in the asymmetric unit.

摘要

最近,从东亚钳蝎(Buthus martensii Karsch,BmK)毒液中纯化出一种对哺乳动物和昆虫均有活性的α-样毒素BmK M7。电生理实验表明,M7可与人心脏钠通道结合并改变其正常特性,因此可被视为一种心脏毒素。采用悬滴气相扩散法,以硫酸铵为沉淀剂,在pH 8.5的Tris-HCl缓冲液中获得了M7的单晶。使用日本光子工厂的同步辐射和电荷耦合器件探测器收集了分辨率为1.40 Å的数据集。数据分析表明,晶体属于空间群P3(1)21/P3(1)21,晶胞参数a = b = 32.76 Å,c = 176.82 Å。假设每个不对称单元中有两个分子,Vm值为1.92 Å3/Da。通过分子置换进行了初步结构分析,结果显示出正确的空间群(P3(1)21)以及不对称单元中两个分子的取向和位置。

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