TY - JOUR
T1 - Nitrosopeptide mapping
T2 - A novel methodology reveals S-nitrosylation of Dexras1 on a single cysteine residue
AU - Jaffrey, Samie R.
AU - Fang, Ming
AU - Snyder, Solomon H.
N1 - Funding Information:
This work was supported NIH grant DA00074 and Research Scientist Award MH-18501 to S.H.S., and MH-66204 and a Young Investigator Award from the National Alliance for Research on Schizophrenia and Depression (NARSAD) to S.R.J.
PY - 2002/12/1
Y1 - 2002/12/1
N2 - 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.
AB - 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.
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U2 - 10.1016/S1074-5521(02)00293-4
DO - 10.1016/S1074-5521(02)00293-4
M3 - Article
C2 - 12498886
AN - SCOPUS:0036918081
SN - 1074-5521
VL - 9
SP - 1329
EP - 1335
JO - Chemistry and Biology
JF - Chemistry and Biology
IS - 12
ER -