期刊
FREE RADICAL BIOLOGY AND MEDICINE
卷 46, 期 10, 页码 1411-1419出版社
ELSEVIER SCIENCE INC
DOI: 10.1016/j.freeradbiomed.2009.02.024
关键词
2,4-Dinitrophenylhydrazine; Protein carbonyls; Dinitrophenyl hydrazone adducts; Cysteine sulfenic acid; Cysteine sulfinic acid; Cysteine sulfonic acid; Thioaldehyde; Hydrogen peroxide; Mass spectrometry; Free radicals
资金
- FIRST (Fondo Interno Ricerca Scientifica e Tecnologica)
- University of Milan
- Fondazione Ariel
- Centro per le Disabilita Neuromotorie Infantili, Milan, Italy
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.
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