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

+ See all authors and affiliations

Science's STKE  12 Jun 2001:
Vol. 2001, Issue 86, pp. pl1
DOI: 10.1126/stke.2001.86.pl1

You are currently viewing the abstract.

View Full Text


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.

View Full Text