Detection and Analysis of Protein S-nitrosylation in Plants

doi:10.11983/CBB19108

Abstract

Abstract: S-nitrosylation is an important protein posttranslational modification, involved in covalently linking a nitric oxide (NO) molecule to the thiol group of a cysteine residue to generate S-nitrosothiols. S-nitrosylation regulates multiple biological processes by modulating protein activity, stability, subcellular localization and protein-protein interactions. The biotin-switch assay is one of the most-often used methods to detect and analyze protein S-nitrosylation. In principle, the free thiols in a target protein are first blocked, followed by reducing the S-nitrosothiols of the target protein to free thiols by ascorbate, which are subsequently labelled by biotin to form biotinylated proteins. The biotin-labelled sample was assayed by immunoblotting and mass spectrometry. Here, we present detailed experimental procedures for the in vitro and in vivo biotin-switch methods and give advice on key troubleshooting solutions.

Key words: nitric oxide, S-nitrosylation, biotin-switch assay, plants

Lichao Chen,Ni Zhan,Yansha Li,Jian Feng,Jianru Zuo. Detection and Analysis of Protein S-nitrosylation in Plants[J]. Chinese Bulletin of Botany, 2019, 54(4): 497-502.

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URL: https://www.chinbullbotany.com/EN/10.11983/CBB19108

https://www.chinbullbotany.com/EN/Y2019/V54/I4/497

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References 28

[1]	Albertos P, Romero-Puertas MC, Tatematsu K, Mateos I, Sánchez-Vicente I, Nambara E, Lorenzo O ( 2015). S-nitrosylation triggers ABI5 degradation to promote seed germination and seedling growth. Nat Commun 6, 8669.
[2]	Chen R, Sun S, Wang C, Li Y, Liang Y, An F, Li C, Dong H, Yang X, Zhang J, Zuo J ( 2009). The Arabidopsis PARAQUAT RESISTANT2 gene encodes an S-nitrosoglutathione reductase that is a key regulator of cell death. Cell Res 19, 1377.
[3]	Cui B, Pan Q, Clarke D, Villarreal MO, Umbreen S, Yuan B, Shan W, Jiang J, Loake GJ ( 2018). S-nitrosylation of the zinc finger protein SRG1 regulates plant immunity. Nat Commun 9, 4226.
[4]	Feng J, Chen L, Zuo J ( 2019). Protein S-nitrosylation in plants: current progresses and challenges. J Integr Plant Biol doi. org/10.1111/jipb.12780.
[5]	Feng J, Wang C, Chen Q, Chen H, Ren B, Li X, Zuo J ( 2013). S-nitrosylation of phosphotransfer proteins represses cytokinin signaling. Nat Commun 4, 1529.
[6]	He Y, Tang RH, Hao Y, Stevens RD, Cook CW, Ahn SM, Jing L, Yang Z, Chen L, Guo F, Fiorani F, Jackson RB, Crawford NM, Pei ZM ( 2004). Nitric oxide represses the Arabidopsis floral transition. Science 305, 1968-1971.
[7]	Hess DT, Matsumoto A, Kim SO, Marshall HE, Stamler JS ( 2005). Protein S-nitrosylation: purview and parameters. Nat Rev Mol Cell Biol 6, 150-166.
[8]	Hess DT, Stamler JS ( 2012). Regulation by S-nitrosylation of protein post-translational modification. J Biol Chem 287, 4411-4418.
[9]	Hu J, Huang X, Chen L, Sun X, Lu C, Zhang L, Wang Y, Zuo J ( 2015). Site-specific nitrosoproteomic identification of endogenously S-nitrosylated proteins in Arabidopsis. Plant Physiol 167, 1731-1746.
[10]	Hu J, Yang H, Mu J, Lu T, Peng J, Deng X, Kong Z, Bao S, Cao X, Zuo J ( 2017). Nitric oxide regulates protein methylation during stress responses in plants. Mol Cell 67, 702-710.
[11]	Iglesias MJ, Terrile MC, Correa-Aragunde N, Colman SL, Izquierdo-Álvarez A, Fiol DF, París R, Sánchez-López N, Marina A, Calderón Villalobos LIA, Estelle M, Lamattina L, Martínez-Ruiz A, Casalongué CA ( 2018). Regulation of SCFTIR1/AFBs E3 ligase assembly by S-nitrosylation of Arabidopsis SKP1-like1 impacts on auxin signaling. Redox Biol 18, 200-210.
[12]	Jaffrey SR, Snyder SH ( 2001). The biotin switch method for the detection of S-nitrosylated proteins. Sci STKE 2001,pl1.
[13]	Ling T, Bellin D, Vandelle E, Imanifard Z, Delledonne M ( 2017). Host-mediated S-nitrosylation disarms the bacterial effector HopAI1 to reestablish immunity. Plant Cell 29, 2871-2881.
[14]	Lytvyn DI, Raynaud C, Yemets AI, Bergounioux C, Blume YB ( 2016). Involvement of inositol biosynthesis and nitric oxide in the mediation of UV-B induced oxidative stress. Front Plant Sci 7, 430.
[15]	Pan QN, Geng CC, Li DD, Xu SW, Mao DD, Umbreen S, Loake GJ, Cui BM ( 2019). Nitrate reductase-mediated nitric oxide regulates the leaf shape in Arabidopsis by mediating the homeostasis of reactive oxygen species. Int J Mol Sci 20, 2235.
[16]	París R, Vazquez MM, Graziano M, Terrile MC, Miller ND, Spalding EP, Otegui MS, Casalongué CA ( 2018). Distribution of endogenous NO regulates early gravitropic response and PIN2 localization in Arabidopsis roots. Front Plant Sci 9, 495.
[17]	Seth D, Hess DT, Hausladen A, Wang L, Wang YJ, Stamler JS ( 2018). A multiplex enzymatic machinery for cellular protein S-nitrosylation. Mol Cell 69, 451-464.
[18]	Seth P, Hsieh PN, Jamal S, Wang L, Gygi SP, Jain MK, Coller J, Stamler JS ( 2019). Regulation of microRNA machinery and development by interspecies S-nitrosylation. Cell 176, 1014-1025.
[19]	Shekariesfahlan A, Lindermayr C ( 2018). Identification of NO-sensitive cysteine residues using cysteine mutants of recombinant proteins. In: Mengel A, Lindermayr C, eds. Nitric Oxide: Methods and Protocols. New York: Springer New York. pp. 183-203.
[20]	Shi H, Liu W, Wei Y, Ye T ( 2017). Integration of auxin/ indole-3-acetic acid 17 and RGA-LIKE3 confers salt stress resistance through stabilization by nitric oxide in Arabidopsis. J Exp Bot 68, 1239-1249.
[21]	Tada Y, Spoel SH, Pajerowska-Mukhtar K, Mou Z, Song J, Wang C, Zuo J, Dong X ( 2008). Plant immunity requires conformational charges of NPR1 via S-nitrosylation and thioredoxins. Science 321, 952-956.
[22]	Wang P, Zhu JK, Lang Z ( 2015). Nitric oxide suppresses the inhibitory effect of abscisic acid on seed germination by S-nitrosylation of SnRK2 proteins. Plant Signal Behav 10, e1031939.
[23]	Willems P, Horne A, Van Parys T, Goormachtig S, De Smet I, Botzki A, Van Breusegem F, Gevaert K ( 2019). The Plant PTM Viewer, a central resource for exploring plant protein modifications. Plant J 99, 752-762.
[24]	Xue Y, Liu Z, Gao X, Jin C, Wen L, Yao X, Ren J ( 2010). GPS-SNO: computational prediction of protein S-nitrosylation sites with a modified GPS algorithm. PLoS One 5, e11290.
[25]	Yang H, Mu J, Chen L, Feng J, Hu J, Li L, Zhou JM, Zuo J ( 2015). S-nitrosylation positively regulates ascorbate peroxidase activity during plant stress responses. Plant Physiol 167, 1604.
[26]	Zhan N, Wang C, Chen L, Yang H, Feng J, Gong X, Ren B, Wu R, Mu J, Li Y, Liu Z, Zhou Y, Peng J, Wang K, Huang X, Xiao S, Zuo J ( 2018). S-nitrosylation targets GSNO reductase for selective autophagy during hypoxia responses in plants. Mol Cell 71, 142-154.
[27]	Zhang L, Shi X, Zhang Y, Wang J, Yang J, Ishida T, Jiang W, Han X, Kang J, Wang X, Pan L, Lv S, Cao B, Zhang Y, Wu J, Han H, Hu Z, Cui L, Sawa S, He J, Wang G ( 2019 a). CLE9 peptide-induced stomatal closure is mediated by abscisic acid, hydrogen peroxide, and nitric oxide in Arabidopsis thaliana. Plant Cell Environ 42, 1033-1044.
[28]	Zhang ZW, Fu YF, Zhou YH, Wang CQ, Lan T, Chen GD, Zeng J, Chen YE, Yuan M, Yuan S, Hu JY ( 2019 b). Nitrogen and nitric oxide regulate Arabidopsis flowering differently. Plant Sci 284, 177-184.