Identification of S-Nitrosothiols by the Sequential Cysteine Blocking Technique

Part of the Methods in Molecular Biology book series (MIMB, volume 1424)


Here, we describe a procedure for the identification of S-nitrosothiols that has been used in our laboratory to study the roles of protein S-nitrosylation in the immune responses of Arabidopsis thaliana and other organisms. It employs a modified version of the biotin-switch technique, which we termed the sequential cysteine blocking technique, encompassing the sequential redox-blocking of recombinant proteins followed by LC–MS/MS analysis.

Key words

S-nitrosylation S-nitrosothiols Redox-based posttranslational modification LC–MS/MS Iodoacetamide Sequential cysteine blocking Label-free quantitation 


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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Institute of Molecular Plant Sciences, School of Biological SciencesUniversity of EdinburghEdinburghUK
  2. 2.Synthetic and Systems Biology, School of Biological SciencesUniversity of EdinburghEdinburghUK

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