Selective fluorescent labeling of S-nitrosothiols (S-FLOS): A novel method for studying S-nitrosation

Lakshmi Santhanam, Marjan Gucek, Tashalee R. Brown, Malini Mansharamani, Sungwoo Ryoo, Christopher A. Lemmon, Lewis Romer, Artin A. Shoukas, Dan E. Berkowitz, Robert N. Cole

Research output: Contribution to journalArticlepeer-review

Abstract

Protein S-nitrosation is a reversible post-translation modification critical for redox-sensitive cell signaling that is typically studied using the Biotin Switch method. This method and subsequent modifications usually require avidin binding or Western blot analysis to detect biotin labeled proteins. We describe here a modification of the Biotin Switch assay that eliminates the need for Western blot or avidin enrichment protocols and allows direct comparison of the S-nitrosation state proteins from two different samples in the same gel lane or on the same 2D gel. This S-FLOS method offers detection, identification and quantification of S-nitrosated proteins, with the potential for site-specific identification of nitrosation events.

Original languageEnglish (US)
Pages (from-to)295-302
Number of pages8
JournalNitric Oxide - Biology and Chemistry
Volume19
Issue number3
DOIs
StatePublished - Nov 2008

Keywords

  • Biotin switch
  • Nitric oxide
  • Proteomics
  • S-nitrosylation
  • SNO

ASJC Scopus subject areas

  • Biochemistry
  • Physiology
  • Clinical Biochemistry
  • Cancer Research

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