title: Formation of protein S-nitrosylation by reactive oxygen species.
abstract: In the present study, the formation of whole cellular S-nitrosylated proteins (protein-SNOs) by the reactive oxygen species (ROS), hydrogen peroxide (H2O2), and superoxide (O2(•-)) is demonstrated. A spectrum of protein cysteine oxidative modifications was detected upon incubation of serum-starved mouse embryonic fibroblasts with increasing concentrations of exogenous H2O2, ranging from exclusive protein-SNOs at low concentrations to a mixture of protein-SNOs and other protein oxidation at higher concentrations to exclusively non-SNO protein oxidation at the highest concentrations of the oxidant used. Furthermore, formation of protein-SNOs was also detected upon inhibition of the antioxidant protein Cu/Zn superoxide dismutase that results in an increase in intracellular concentration of O2(•-). These results were further validated using the phosphatase and tensin homologue, PTEN, as a model of a protein sensitive to oxidative modifications. The formation of protein-SNOs by H2O2 and O2(•-) was prevented by the NO scavenger, c-PTIO, as well as the peroxinitrite decomposition catalyst, FETPPS, and correlated with the production or the consumption of nitric oxide (NO), respectively. These data suggest that the formation of protein-SNOs by H2O2 or O2(•-) requires the presence or the production of NO and involves the formation of the nitrosylating intermediate, peroxinitrite.
authors.affiliation: Department of Biochemistry and National University of Singapore Graduate School for Integrative Sciences and Engineering, National University of Singapore, Yong Loo Lin School of Medicine , Singapore.
authors.forename: K Htet
authors.initials: KH
authors.lastname: Hlaing
authors.forename: M-V
authors.initials: MV
authors.lastname: Clément
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002478:Cells, Cultured; D005347:Fibroblasts / Q000378:metabolism; D051379:Mice; D009569:Nitric Oxide / Q000378:metabolism; D010084:Oxidation-Reduction; D011506:Proteins / Q000378:metabolism*; D017382:Reactive Oxygen Species / Q000378:metabolism*
chemical_list: D011506:Proteins; D017382:Reactive Oxygen Species; D009569:Nitric Oxide
keywords: hydrogen peroxide; nitric oxide; nitrosylation; peroxinitrite; redox signaling; superoxide