Protein carbonylation: 2,4-dinitrophenylhydrazine reacts with both aldehydes/ketones and sulfenic acids
Articolo
Data di Pubblicazione:
2009
Citazione:
Protein carbonylation: 2,4-dinitrophenylhydrazine reacts
with both aldehydes/ketones and sulfenic acids / I. Dalle-Donne, M. Carini, M. Orioli, G. Vistoli, L. Regazzoni, G. Colombo, R. Rossi, A. Milzani, G. Aldini. - In: FREE RADICAL BIOLOGY & MEDICINE. - ISSN 0891-5849. - 46:10(2009), pp. 1411-1419.
Abstract:
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. β-Mercaptoethanol prevents the formation of the DNPH derivative because it reacts with the oxidized Cys residue, forming the corresponding disulfide.
Tipologia IRIS:
01 - Articolo su periodico
Keywords:
2,4-Dinitrophenylhydrazine; Cysteine sulfenic acid; Cysteine sulfinic acid; Cysteine sulfonic acid; Dinitrophenyl hydrazone adducts; Free radicals; Hydrogen peroxide; Mass spectrometry; Protein carbonyls; Thioaldehyde
Elenco autori:
I. Dalle-Donne, M. Carini, M. Orioli, G. Vistoli, L. Regazzoni, G. Colombo, R. Rossi, A. Milzani, G. Aldini
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