生物素化
S-亚硝基化
半胱氨酸
化学
亲和素
一氧化氮
生物化学
加合物
亲和层析
体外
生物素
酶
有机化学
作者
Samie R. Jaffrey,Solomon H. Snyder
出处
期刊:Science's STKE
[American Association for the Advancement of Science (AAAS)]
日期:2001-06-12
卷期号:2001 (86)
被引量:616
标识
DOI:10.1126/stke.2001.86.pl1
摘要
Many of the effects of nitric oxide are mediated by the direct modification of cysteine residues resulting in an adduct called a nitrosothiol. Here, we describe a novel method for detecting proteins that contain nitrosothiols. In this three-step procedure, nitrosylated cysteines are converted to biotinylated cysteines. Biotinylated proteins can then be detected by immunoblotting or can be purified by avidin-affinity chromatography. We include examples of the detection of S-nitrosylated proteins in brain lysates after in vitro S-nitrosylation, as well as the detection of endogenous S-nitrosothiols in selected neuronal proteins.
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