Affordable Access

deepdyve-link
Publisher Website

Rocket fuel for the quantification of S-nitrosothiols. Highly specific reduction of S-nitrosothiols to thiols by methylhydrazine.

Authors
  • Wiesweg, M
  • Berchner-Pfannschmidt, U
  • Fandrey, J
  • Petrat, F
  • de Groot, H
  • Kirsch, M
Type
Published Article
Journal
Free Radical Research
Publisher
Informa UK (Taylor & Francis)
Publication Date
Feb 01, 2013
Volume
47
Issue
2
Pages
104–115
Identifiers
DOI: 10.3109/10715762.2012.744836
PMID: 23181469
Source
Medline
License
Unknown

Abstract

Reduction of S-nitrosothiols to the corresponding thiol function is the key step in analyzing S-nitrosocysteinyl residues in proteins. Though it has been shown to give low yields, ascorbate-dependent reduction is commonly performed in the frequently used biotin-switch technique. We demonstrate that the compound methylhydrazine can act as a specific and efficient reducing agent for S-nitrosothiols. The corresponding thiol function is exclusively generated from low molecular weight and proteinaceous S-nitrosothiols while methylhydrazine failed to reduce disulfides. It was possible to optimize the experimental conditions so that thiol autoxidation is excluded, and high reaction yields (>90%) are obtained for the thiol function. The biotin-switch technique performed with methylhydrazine-dependent reduction shows remarkably improved sensitivity compared to the ascorbate-dependent procedure.

Report this publication

Statistics

Seen <100 times