Nitrosopeptide mapping: A novel methodology reveals S-nitrosylation of Dexras1 on a single cysteine residue

Samie R. Jaffrey, Ming Fang, Solomon H Snyder

Research output: Contribution to journalArticle

Abstract

S-Nitrosylation of specific cysteine residues is a reversible signaling mechanism of nitric oxide (NO) generated by NO synthase (NOS) enzymes. In some proteins, evidence has accumulated that more than one cysteine can be S-nitrosylated; however, it is difficult to distinguish S-nitrosylation on separate cysteine residues. We report a novel simple, sensitive, and specific procedure for nitrosopeptide mapping. Dexras1 is a monomeric G protein whose guanine nucleotide exchange activity is augmented by NO; the identity and number of its S-nitrosylated cysteines is unknown. We describe the radiolabeling of S-nitrosylated cysteine residues in Dexras1. A nitrosopeptide map, generated by two-dimensional peptide chromatography, reveals that only a single cysteine is S-nitrosylated following NO exposure. Mutagenesis of Cys11 abolished the effect of NO donors on Dexras1, implicating this residue in the NO-mediated activation of Dexras1.

Original languageEnglish (US)
Pages (from-to)1329-1335
Number of pages7
JournalChemistry and Biology
Volume9
Issue number12
DOIs
StatePublished - Dec 1 2002

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Cysteine
Nitric Oxide
Mutagenesis
Guanine Nucleotides
Monomeric GTP-Binding Proteins
Nitric Oxide Donors
Chromatography
Nitric Oxide Synthase
Chemical activation
Peptides
Enzymes
Proteins

ASJC Scopus subject areas

  • Organic Chemistry

Cite this

Nitrosopeptide mapping : A novel methodology reveals S-nitrosylation of Dexras1 on a single cysteine residue. / Jaffrey, Samie R.; Fang, Ming; Snyder, Solomon H.

In: Chemistry and Biology, Vol. 9, No. 12, 01.12.2002, p. 1329-1335.

Research output: Contribution to journalArticle

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