Resin-assisted enrichment of thiols as a general strategy for proteomic profiling of cysteine-based reversible modifications.

ABSTRACT: Reversible modifications of cysteine thiols have a key role in redox signaling and regulation. A number of reversible redox modifications, including disulfide formation, S-nitrosylation (SNO) and S-glutathionylation (SSG), have been recognized for their significance in various physiological and pathological processes. Here we describe a procedure for the enrichment of peptides containing reversible cysteine modifications. Starting with tissue or cell lysate samples, all of the unmodified free thiols are blocked using N-ethylmaleimide (NEM). This is followed by the selective reduction of those cysteines bearing the reversible modification(s) of interest. The reduction is achieved by using different reducing reagents that react specifically with each type of cysteine modification (e.g., asco

SUBMITTER: Guo J 

PROVIDER: S-EPMC4038159 | biostudies-literature | 2014 Jan

REPOSITORIES: biostudies-literature