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Fluorimetric-Based Method to Detect and Quantify Total S-Nitrosothiols (SNOs) in Plant Samples

  • Paulo T. MiotoEmail author
  • Alejandra Matiz
  • Luciano Freschi
  • Francisco J. Corpas
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2057)

Abstract

Accumulating experimental evidence indicates that S-nitrosylation (technically S-nitrosation) events have a central role in plant biology, presumably accounting for much of the widespread influence of nitric oxide (NO) on developmental, metabolic, and stress-related plant responses. Therefore, the accurate detection and quantification of S-nitrosylated proteins and peptides can be particularly useful to determine the relevance of this class of compounds in the ever-increasing number of NO-dependent signaling events described in plant systems. Up to now, the quantification of S-nitrosothiols (SNOs) in plant samples has mostly relied on the Saville reaction and the ozone-based chemiluminescence method, which lacks sensitivity and are very time-consuming, respectively. Taking advantage of the photolytic properties of S-nitrosylated proteins and peptides, the method described in this chapter allows simple, fast, and high-throughput detection of SNOs in plant samples.

Key words

Ozone chemiluminescence Saville S-nitrosothiols S-nitrosoglutathione Nitric oxide Nitrosylated proteins 

Notes

Acknowledgments

Funding: This work was supported by Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) [grant numbers 2011/50637-0, 2013/18056-2 and 2013/15108-1], Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) [grant numbers 442045/2014-0, and 309504/2014-7], Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) [grant number 99999.006262/2015-05] and CNCSIS/UEFISCDI [grant PN-II-RU-TE-2014-4-2555]. Research in FJC laboratory is supported by the ERDF-cofinanced grants from the Ministry of Economy and Competitiveness (AGL2015-65104-P) and Junta de Andalucía (group BIO192), Spain.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Paulo T. Mioto
    • 1
    Email author
  • Alejandra Matiz
    • 2
  • Luciano Freschi
    • 2
  • Francisco J. Corpas
    • 3
  1. 1.Department of Botany, Biological Sciences CenterUniversidade Federal de Santa CatarinaFlorianópolisBrazil
  2. 2.Department of Botany, Institute of BiosciencesUniversity of São PauloSão PauloBrazil
  3. 3.Department of Biochemistry, Cell and Molecular Biology of PlantsEstación Experimental del Zaidín, CSICGranadaSpain

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