Nitrosopeptide mapping: a novel methodology reveals s-nitrosylation of dexras1 on a single cysteine residue. Academic Article uri icon

Overview

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.

publication date

  • December 1, 2002

Research

keywords

  • Cysteine
  • GTP-Binding Proteins
  • Monomeric GTP-Binding Proteins
  • Peptide Mapping
  • S-Nitrosothiols
  • ras Proteins

Identity

Scopus Document Identifier

  • 0036918081

Digital Object Identifier (DOI)

  • 10.1016/s1074-5521(02)00293-4

PubMed ID

  • 12498886

Additional Document Info

volume

  • 9

issue

  • 12