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

S. R. Jaffrey, S. H. Snyder

Research output: Contribution to journalArticle

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.

Original languageEnglish (US)
Pages (from-to)PL1
JournalScience's STKE : signal transduction knowledge environment
Volume2001
Issue number86
DOIs
StatePublished - Jun 12 2001

ASJC Scopus subject areas

  • Medicine(all)

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