Method for assaying protein nitrosylation

US6806057B2 · kind B2 · utility

Assignee

• THE JOHNS HOPKINS UNIVERSITY · US

Inventors

• Solomon H. Snyder · Baltimore, US
• Samie R. Jaffrey · New York, US
• Christopher Ferris · Franklin, US
• Hediye Erdjument-Bromage · New York, US
• Paul Tempst · New York, US

Key dates

• Filing date: Oct 29, 2001
• Grant date: Oct 19, 2004
• Expiry date: Oct 29, 2021

Classification

• Technology area (CPC Y): Emerging Cross-Sectional Technologies
• CPC primary: Y10T436/177692
• WIPO field: Measurement
• WIPO sector: Instruments

Abstract

Many of the effects of nitric oxide are mediated by the direct modification of cysteine residues resulting in an adduct called a nitrosothiol. A method to detect proteins which contain nitrosothiols involves several steps. Nitrosylated cysteines are converted to tagged cysteines. Tagged proteins can then be detected, for example, by immunoblotting and/or can be purified by affinity chromatography. The method is applicable to the detection of S-nitrosylated proteins in cell lysates following in vitro S-nitrosylation, as well as to the detection of endogenous S-nitrosothiols in selected protein substrates.