Abstract
S-nitrosation as a redox-based posttranslational modification of protein cysteine has emerged as an integral part of signaling pathways of nitric oxide across all types of organisms. Protein S-nitrosation status is controlled by two key mechanisms: by direct denitrosation performed by the thioredoxin/thioredoxin reductase system, and in an indirect way mediated by S-nitrosoglutathione reductase (GSNOR). GSNOR, which has been identified as a key component of S-nitrosothiols catabolism, catalyzes an irreversible decomposition of abundant intracellular S-nitrosothiol, S-nitrosoglutathione (GSNO) to oxidized glutathione using reduced NADH cofactor. In plants, GSNOR has been shown to play important roles in plant growth and development and plant responses to abiotic and biotic stress stimuli. In this chapter, optimized protocols of spectrophotometric measurement of GSNOR enzymatic activity and activity staining in native polyacrylamide gels in plant GSNOR are presented.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Yu M, Lamattina L, Spoel SH, Loake GJ (2014) Nitric oxide function in plant biology: a redox cue in deconvolution. New Phytol 202:1142–1156
Seth D, Stamler JS (2011) The SNO-proteome: causation and classifications. Curr Opin Chem Biol 15:129–136
Martínez-Ruiz A, Lamas S (2004) S-nitrosylation: a potential new paradigm in signal transduction. Cardiovasc Res 62:43–52
Corpas FJ, Alché JD, Barroso JB (2013) Current overview of S-nitrosoglutathione (GSNO) in higher plants. Frontiers Plant Sci 4:126
Benhar M, Forrester MT, Stamler JS (2009) Protein denitrosylation: enzymatic mechanisms and cellular functions. Nat Rev Mol Cell Biol 10:721–732
Koivusalo M, Baumann M, Uotila L (1989) Evidence for the identity of glutathione-dependent formaldehyde dehydrogenase and class III alcohol dehydrogenase. FEBS Lett 257:105–109
Jensen D, Belka G, Du Bois G (1998) S-Nitrosoglutathione is a substrate for rat alcohol dehydrogenase class III isoenzyme. Biochem J 331:659–668
Liu L, Hausladen A, Zeng M et al (2001) A metabolic enzyme for S-nitrosothiol conserved from bacteria to humans. Nature 410:490–494
Staab CA, Hellgren M, Höög JO (2008) Medium- and short-chain dehydrogenase/reductase gene and protein families: dual functions of alcohol dehydrogenase 3: implications with focus on formaldehyde dehydrogenase and S-nitrosoglutathione reductase activities. Cell Mol Life Sci 65:3950–3960
Kubienová L, Kopečný D, Tylichová M et al (2013) Structural and functional characterization of a plant S-nitrosoglutathione reductase from Solanum lycopersicum. Biochimie 95:889–902
Leterrier M, Chaki M, Airaki M et al (2011) Function of S-nitrosoglutathione reductase (GSNOR) in plant development and under biotic/abiotic stress. Plant Signal Behav 6:789–793
Xu S, Guerra D, Lee U, Vierling E (2013) S-nitrosoglutathione reductases are low-copy number, cysteine-rich proteins in plants that control multiple developmental and defense responses in Arabidopsis. Frontiers Plant Sci 4:430
Corpas FJ, Carreras A, Esteban FJ et al (2008) Localization of S-nitrosothiols and assay of nitric oxide synthase and S-nitrosoglutathione reductase activity in plants. Methods Enzymol 437:561–574
Chaki M, Valderrama R, Fernández-Ocaňa AM et al (2011) Mechanical wounding induces a nitrosative stress by down-regulation of GSNO reductase and an increase in S-nitrosothiols in sunflower (Helianthus annuus) seedlings. J Exp Bot 62:1803–1813
Barnett SD, Buxton LO (2017) The role of S-nitrosoglutathione reductase (GSNOR) in human disease and therapy. Crit Rev Biochem Mol Biol 52:340–354
Zor T, Selinger Z (1996) Linearization of the Bradford protein assay increases its sensitivity: theoretical and experimental studies. Anal Biochem 236:302–308
Moore KP, Mani AR (2002) Measurement of protein nitration and S-nitrosothiol formation in biology and medicine. In: Cadenas E, Lester P (eds) Methods in enzymology. Academic Press, Cambridge, MA, pp 256–268
Uotila L, Koivusalo M (1974) Purification and properties of S-formylglutathione hydrolase from human liver. J Biol Chem 249:7664–7672
Acknowledgments
This project was supported by an internal Grant from Palacky University in Olomouc (IGA_PrF_2019_022).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2020 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Janků, M., Tichá, T., Luhová, L., Petřivalský, M. (2020). Measurement of S-Nitrosoglutathione Reductase Activity in Plants. In: Gupta, K. (eds) Nitrogen Metabolism in Plants. Methods in Molecular Biology, vol 2057. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9790-9_5
Download citation
DOI: https://doi.org/10.1007/978-1-4939-9790-9_5
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-4939-9789-3
Online ISBN: 978-1-4939-9790-9
eBook Packages: Springer Protocols