Identification of S-Nitrosothiols by the Sequential Cysteine Blocking Technique

Gary Loake, Rafael A Homem, Thierry Le Bihan, Manda Yu

Research output: Chapter in Book/Report/Conference proceedingChapter (peer-reviewed)peer-review

Abstract

Here, we describe a procedure for the identifi cation 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 modifi ed 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.
Original languageEnglish
Title of host publicationPlant Nitric Oxide
Subtitle of host publicationMethods and Protocols
PublisherSpringer New York
Pages163-174
Number of pages11
Volume1424
ISBN (Electronic)978-1-4939-3600-7
ISBN (Print)978-1-4939-3598-7
DOIs
Publication statusPublished - 20 Apr 2016

Publication series

NameMethods in Molecular Biology
Volume1424
ISSN (Print)1064-3745

Keywords

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

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