用于检测S-亚硝基化蛋白的生物素开关法。

The biotin switch method for the detection of S-nitrosylated proteins.

作者信息

Jaffrey S R, Snyder S H

机构信息

Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA. E-mail:

出版信息

Sci STKE. 2001 Jun 12;2001(86):pl1. doi: 10.1126/stke.2001.86.pl1.

DOI:10.1126/stke.2001.86.pl1

PMID:11752655

Abstract

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.

摘要

一氧化氮的许多作用是通过直接修饰半胱氨酸残基介导的，从而形成一种称为亚硝基硫醇的加合物。在此，我们描述了一种检测含有亚硝基硫醇的蛋白质的新方法。在这个三步程序中，亚硝基化的半胱氨酸被转化为生物素化的半胱氨酸。然后可以通过免疫印迹检测生物素化的蛋白质，或者通过抗生物素蛋白亲和色谱法进行纯化。我们提供了体外亚硝基化后在脑裂解物中检测S-亚硝基化蛋白质的实例，以及在选定的神经元蛋白质中检测内源性S-亚硝基硫醇的实例。