Identification of S-nitrosylated Proteins in Arabidopsis thaliana after Gamma Irradiation
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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.
Abstract
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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