1. Nitrosothiol Reactivity Profiling Identifies S-Nitrosylated Proteins with Unexpected Stability
- Author
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Samie R. Jaffrey, Guoqiang Xu, Jeremy S. Paige, and Branka Stancevic
- Subjects
Proteomics ,Molecular Sequence Data ,Clinical Biochemistry ,Protein Array Analysis ,Nitric Oxide ,Biochemistry ,Gas Chromatography-Mass Spectrometry ,Article ,Nitric oxide ,Mice ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Drug Stability ,Drug Discovery ,Animals ,Humans ,Amino Acid Sequence ,Molecular Biology ,Peptide sequence ,030304 developmental biology ,Pharmacology ,0303 health sciences ,S-Nitrosothiols ,Molecular Structure ,Nitrosylation ,Brain ,General Medicine ,Glutathione ,Rats ,Cytosol ,CHEMBIO ,chemistry ,SIGNALING ,Molecular Medicine ,Signal transduction ,Crystallization ,Peptides ,Oxidation-Reduction ,030217 neurology & neurosurgery ,Chromatography, Liquid ,Signal Transduction ,Cysteine - Abstract
Summary Nitric oxide (NO) regulates protein function by S -nitrosylation of cysteine to form nitrosothiols. Nitrosothiols are highly susceptible to nonenzymatic degradation by cytosolic reducing agents. Here we show that although most protein nitrosothiols are rapidly degraded by cytosolic reductants, a small subset form unusually stable S -nitrosylated proteins. Our findings suggest that stable S -nitrosylation reflects a protein conformation change that shields the nitrosothiol. To identify stable protein nitrosothiols, we developed a proteomic method for profiling S -nitrosylation. We examined the stability of over 100 S -nitrosylated proteins, and identified 10 stable nitrosothiols. These proteins remained S -nitrosylated in cells after NO synthesis was inhibited, unlike most S -nitrosylated proteins. Taken together, our data identify a class of NO targets that form stable nitrosothiols in the cell and are likely to mediate the persistent cellular effects of NO.
- Published
- 2008
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