Journal
FREE RADICAL BIOLOGY AND MEDICINE
Volume 46, Issue 10, Pages 1411-1419Publisher
ELSEVIER SCIENCE INC
DOI: 10.1016/j.freeradbiomed.2009.02.024
Keywords
2,4-Dinitrophenylhydrazine; Protein carbonyls; Dinitrophenyl hydrazone adducts; Cysteine sulfenic acid; Cysteine sulfinic acid; Cysteine sulfonic acid; Thioaldehyde; Hydrogen peroxide; Mass spectrometry; Free radicals
Funding
- FIRST (Fondo Interno Ricerca Scientifica e Tecnologica)
- University of Milan
- Fondazione Ariel
- Centro per le Disabilita Neuromotorie Infantili, Milan, Italy
Ask authors/readers for more resources
Most of the assays for detection of carbonylated proteins, the most general and widely used marker of severe protein oxidation, involve derivatization of the carbonyl group with 2,4-dinitrophenylhydrazine (DNPH), which leads to formation of a stable dinitrophenyl hydrazone product. Here, by using a Cys-containing model peptide and high-resolution mass spectrometry, we demonstrate that DNPH is not exclusively selective for carbonyl groups, because it also reacts with sulfenic acids, forming a DNPH adduct, through the acid-catalyzed formation of a thioaldehyde intermediate that is further converted to an aldehyde. beta-Mercaptoethanol prevents the formation of the DNPH derivative because it reacts with the oxidized Cys residue, forming the corresponding disulfide. (C) 2009 Elsevier Inc. All rights reserved.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available