Selective production of rat mutant selenoprotein W with and without bound glutathione

被引：10

作者：

Bauman, AT

Malencik, DA

Barofsky, DF

Barofsky, E

Anderson, SR

Whanger, PD ^{[1
]}

机构：

[1] Oregon State Univ, Dept Environm & Mol Toxicol, Corvallis, OR 97331 USA

[2] Oregon State Univ, Dept Biochem & Biophys, Corvallis, OR 97331 USA

[3] Oregon State Univ, Dept Chem, Corvallis, OR 97331 USA

来源：

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS | 2004年 / 313卷 / 02期

关键词：

selenoprotein W; selenoproteins; Ni-NTA chromatography; His-tagged proteins; glutathione binding;

D O I：

10.1016/j.bbrc.2003.11.133

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS) and electrospray ionization mass spectrometry (ESI MS) analysis of a 6x His-tagged recombinant form of rat mutant selenoprotein W (RMSW) reveals that aerobic growth conditions primarily produce a form of RMSW without bound glutathione (10,305 Da) whereas anaerobic conditions produce a glutathione-bound (305Da) form (10,610 Da). Purification of RMSW was achieved with a procedure employing acetone precipitation and DEAE-cellulose chromatography, in addition to Ni-NTA agarose chromatography. Additional steps, including polyvalent metal ion binding (PMIB) resin chromatography and CM-cellulose chromatography, were necessary after elution from the Ni-NTA agarose column, in order to maintain solubility of the purified protein. (C) 2003 Elsevier Inc. All rights reserved.

引用

页码：308 / 313

页数：6

共 22 条

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