Protein carbonylation: 2,4-dinitrophenylhydrazine reacts with both aldehydes/ketones and sulfenic acids.
Free Radic Biol Med, 2009/5/15;46(10):1411-9.
Dalle-Donne I[1], Carini M, Orioli M, Vistoli G, Regazzoni L, Colombo G, Rossi R, Milzani A, Aldini G
Affiliations
PMID: 19268703DOI: 10.1016/j.freeradbiomed.2009.02.024
Impact factor: 8.101
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. beta-Mercaptoethanol prevents the formation of the DNPH derivative because it reacts with the oxidized Cys residue, forming the corresponding disulfide.
MeSH terms
Aldehydes; Computational Biology; Cysteine; Dimerization; Ketones; Models, Theoretical; Oxidation-Reduction; Peptides; Phenylhydrazines; Protein Binding; Protein Carbonylation; Sulfenic Acids
More resources
EndNote: Download