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大鼠动脉壁和脑中新发现的含硒蛋白质的纯化。

Purification of the newly found selenium-containing proteins in the arterial wall and brain of the rat.

作者信息

Qu X, Huang K, Wu Z, Zhong S, Chen Z, Xu H

机构信息

Department of Chemistry, Huazhong University of Science and Technology, Wuhan, 430074, People's Republic of China.

出版信息

Biochem Biophys Res Commun. 2000 Apr 21;270(3):688-94. doi: 10.1006/bbrc.2000.2512.

Abstract

Previously several selenium-containing proteins with different subunit molecular masses (M(r)) were detected in the arterial wall and brain of rats. In continuation of this work, after labeling of rats in vivo with [(75)Se]selenite, the new selenium-containing proteins of interest were purified on a Sephadex G-200 column followed by preparative isoelectric focusing. Nuclear analytical methods (gamma-counter and gamma-detector) were applied in the detection and identification of the (75)Se-labeled proteins. The two (75)Se-containing proteins from the arterial wall migrated as 15.0- and 67.0-kDa species on SDS-PAGE gels with pI values of 4.5 and 5.1, respectively. The three (75)Se-containing proteins from brain purified to homogeneity had M(r) values of 18.0, 30.0, and 42.9 kDa and pI values of 6.3, 6.5, and 6.0, respectively. Of these proteins, the 67.0-, 42.9-, and 30.0-kDa species may be yet not characterized selenoproteins with important biological functions.

摘要

此前在大鼠的动脉壁和大脑中检测到了几种具有不同亚基分子量(M(r))的含硒蛋白。在这项工作的后续研究中,用[(75)Se]亚硒酸盐对大鼠进行体内标记后,通过Sephadex G - 200柱纯化目标新含硒蛋白,随后进行制备性等电聚焦。核分析方法(γ计数器和γ探测器)用于检测和鉴定(75)Se标记的蛋白。来自动脉壁的两种含(75)Se蛋白在SDS - PAGE凝胶上迁移时分子量分别为15.0 kDa和67.0 kDa,等电点(pI)值分别为4.5和5.1。从大脑中纯化至均一的三种含(75)Se蛋白的M(r)值分别为18.0、30.0和42.9 kDa,pI值分别为6.3、6.5和6.0。在这些蛋白中,67.0 kDa、42.9 kDa和30.0 kDa的蛋白可能是尚未被表征的具有重要生物学功能的硒蛋白。

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