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Sci STKE. 2001 Jun 12;2001(86):pl1.

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

Author information

1
Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA. E-mail: jaffrey@jhmi.edu

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.

PMID:
11752655
DOI:
10.1126/stke.2001.86.pl1
[Indexed for MEDLINE]

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