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Nitric oxide (NO), an important gaseous signaling molecule and reactive nitrogen species(RNS), regulates several developmental processes and stress responses in plants via proteinS-nitrosylation, in which specific cysteine thiols are converted to S-nitrosothiols (SNOs). However,there is no information available regarding the formation of nitrosothiols in Arabidopsis due to gammaradiation. In this study, S-nitrosylated proteins were detected using the biotin switch method and iodoTMTlabeling. After gamma irradiation, S-nitrosylated proteins increased in a dose-dependent manner. UsingiodoTMT labeling, we identified 178 proteins following a site-specific nitrosoproteomic approach due tothe 250 Gy dose of gamma irradiation. The identified proteins belonged to different functional families,including photosynthesis (19.9%), metabolism (10.8%), redox processes (10.2%), protein processing (10.2%),protein synthesis (10.2%), miscellaneous (6.3%), and others. Among them, we selected high-scoringproteins, such as AtPrx and AtTDX, and verified their S-nitrosylation through two purified proteins byimmunoblotting. This study will be beneficial in understanding the biological effects of radiation.
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