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Nitric Oxide pp 267–280Cite as

Immunodetection of S-Nitrosoglutathione Reductase Protein in Plant Samples

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1747))

Abstract

S-nitrosation, the attachment of a nitroso group to cysteine thiols, has been recognized as an important posttranslational modification of proteins by nitric oxide and related reactive nitrogen species. Mechanisms and significance of S-nitrosation in the regulation of the structure and activity of proteins have been extensively studied in animal and plant systems. In plants, protein S-nitrosation is involved in signaling pathways of plant hormones and regulators during plant growth and development and in responses to abiotic and biotic stress stimuli. S-nitrosoglutathione reductase (GSNOR) has been identified as a key enzyme controlling the intracellular level of S-nitrosothiols. GSNOR irreversibly degrades S-nitrosoglutathione (GSNO), the major low molecular weight S-nitrosothiol involved in the formation of protein S-nitrosothiols through transnitrosylation. GSNOR level and activity in plant cells are modulated during plant development and in response to external stimuli such as pathogen infection. In this chapter, we give a detailed description of the immunochemical detection of the GSNOR protein in plant samples.

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References

  1. Gould N, Doulias PT, Tenopoulou M, Raju K, Ischiropoulos H (2013) Regulation of protein function and signaling by reversible cysteine S-nitrosylation. J Biol Chem 288:26473–26479

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Lamotte O, Bertoldo JB, Besson-Bard A, Rosnoblet C, Aimé S et al (2015) Protein S-nitrosylation: specificity and identification strategies in plants. Front Chem 2:114

    Article  PubMed  PubMed Central  Google Scholar 

  3. Kovacs I, Lindermayr C (2013) Nitric oxide-based protein modification: formation and site-specificity of protein S-nitrosylation. Front Plant Sci 4:137

    PubMed  PubMed Central  Google Scholar 

  4. 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

    Article  CAS  PubMed  Google Scholar 

  5. Liu L, Hausladen A, Zeng M, Que L, Heitman J et al (2001) A metabolic enzyme for S-nitrosothiol conserved from bacteria to humans. Nature 410:490–494

    Article  CAS  PubMed  Google Scholar 

  6. Staab CA, Ålander J, Brandt M, Lengqvist J, Morgenstern R et al (2008) Reduction of S-nitrosoglutathione by alcohol dehydrogenase 3 is facilitated by substrate alcohols via direct cofactor recycling and leads to GSH-controlled formation of glutathione transferase inhibitors. Biochem J 413:493–504

    Article  CAS  PubMed  Google Scholar 

  7. 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. Front Plant Sci 4:430

    Article  PubMed  PubMed Central  Google Scholar 

  8. Salgado I, Carmen Martínez M, Oliveira H, Frungillo L (2013) Nitric oxide signaling and homeostasis in plants: a focus on nitrate reductase and S-nitrosoglutathione reductase in stress-related responses. Braz J Bot 36:89–98

    Article  Google Scholar 

  9. Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A 76:4350–4354

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Espunya MC, Díaz M, Moreno-Romero J, Martínez MC (2006) Modification of intracellular levels of glutathione-dependent formaldehyde dehydrogenase alters glutathione homeostasis and root development. Plant Cell Environ 29:1002–1011

    Article  CAS  PubMed  Google Scholar 

  11. Kubienova L, Kopecny D, Tylichova M, Briozzo P, Skopalova J et al (2013) Structural and functional characterization of a plant S-nitrosoglutathione reductase from Solanum lycopersicum. Biochimie 95:889–902

    Article  CAS  PubMed  Google Scholar 

  12. Kubienova L, Ticha T, Jahnova J, Luhova L, Mieslerova B et al (2014) Effect of abiotic stress stimuli on S-nitrosoglutathione reductase in plants. Planta 239:139–146

    Article  CAS  PubMed  Google Scholar 

  13. Tichá T, Činčalová L, Kopečný D, Sedlářová M, Kopečná M et al (2017) Characterization of S-nitrosoglutathione reductase from Brassica and Lactuca spp. and its modulation during plant development. Nitric Oxide 68:68–76

    Google Scholar 

  14. Chaki M, Fernández-Ocaňa AM, Valderrama R, Carreras A, Esteban FJ et al (2009) Involvement of reactive nitrogen and oxygen species (RNS and ROS) in sunflower-mildew interaction. Plant Cell Physiol 50:265–279

    Article  CAS  PubMed  Google Scholar 

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Correspondence to Marek Petřivalský .

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Tichá, T., Luhová, L., Petřivalský, M. (2018). Immunodetection of S-Nitrosoglutathione Reductase Protein in Plant Samples. In: Mengel, A., Lindermayr, C. (eds) Nitric Oxide. Methods in Molecular Biology, vol 1747. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7695-9_21

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  • DOI: https://doi.org/10.1007/978-1-4939-7695-9_21

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7694-2

  • Online ISBN: 978-1-4939-7695-9

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