doi: 10.1126/stke.2001.86.pl1.
The biotin switch method for the detection of S-nitrosylated proteins
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- PMID: 11752655
- DOI: 10.1126/stke.2001.86.pl1
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The biotin switch method for the detection of S-nitrosylated proteins
Sci STKE.
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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.
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