Formation of protein S-nitrosylation by reactive oxygen species

• Published in: Free radical research Vol. 48; no. 9; pp. 996 - 1010
• Main Authors: Htet Hlaing, K., Clément, M.-V.
• Format: Journal Article
• Language: English
• Published: England Informa Healthcare 01.09.2014; Taylor & Francis
• Subjects: Animals; Blotting, Western; Cells, Cultured; Fibroblasts - metabolism; hydrogen peroxide; Mice; nitric oxide; Nitric Oxide - metabolism; nitrosylation; Oxidation-Reduction; peroxinitrite; Proteins - metabolism; Reactive Oxygen Species - metabolism; redox signaling; superoxide; nitrosylation; nitric oxide; redox signaling; peroxinitrite; superoxide; hydrogen peroxide
• ISSN: 1071-5762; 1029-2470
• DOI: 10.3109/10715762.2014.942842

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.