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Sci. STKE, 12 June 2001
Vol. 2001, Issue 86, p. pl1
[DOI: 10.1126/scisignal.862001pl1]

PROTOCOLS

The Biotin Switch Method for the Detection of S-Nitrosylated Proteins

Samie R. Jaffrey and Solomon H. Snyder

The authors are in the Departments of Neuroscience, Pharmacology and Molecular Sciences, and Psychiatry, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA. E-mail: jaffrey{at}jhmi.edu; ssnyder{at}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.

Citation: S. R. Jaffrey, S. H. Snyder, The Biotin Switch Method for the Detection of S-Nitrosylated Proteins. Sci. STKE 2001, pl1 (2001).

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