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.
|Titolo:||Protein carbonylation: 2,4-dinitrophenylhydrazine reacts with both aldehydes/ketones and sulfenic acids.|
|Rivista:||FREE RADICAL BIOLOGY & MEDICINE|
|Citazione:||DALLE DONNE, I., Carini, M., Orioli, M., Vistoli, G., Regazzoni, L., Colombo, G., et al. (2009). Protein carbonylation: 2,4-dinitrophenylhydrazine reacts with both aldehydes/ketones and sulfenic acids. FREE RADICAL BIOLOGY & MEDICINE, 46(10), 1411-1419.|
|Appare nelle tipologie:||1.1 Articolo in rivista|