Mechanism of protein decarbonylation.
Ligand/receptor stimulation of cells promotes protein carbonylation that is followed by the decarbonylation process, which might involve thiol-dependent reduction (C.M. Wong et al., Circ. Res. 102:301-318; 2008). This study further investigated the properties of this protein decarbonylation mechanism. We found that the thiol-mediated reduction of protein carbonyls is dependent on heat-labile biologic components. Cysteine and glutathione were efficient substrates for decarbonylation. Thiols decreased the protein carbonyl content, as detected by 2,4-dinitrophenylhydrazine, but not the levels of malondialdehyde or 4-hydroxynonenal protein adducts. Mass spectrometry identified proteins that undergo thiol-dependent decarbonylation, which include peroxiredoxins. Peroxiredoxin-2 and -6 were carbonylated and subsequently decarbonylated in response to the ligand/receptor stimulation of cells. siRNA knockdown of glutaredoxin inhibited the decarbonylation of peroxiredoxin. These results strengthen the concept that thiol-dependent decarbonylation defines the kinetics of protein carbonylation signaling.
['Wong|Chi-Ming|CM|', 'Marcocci|Lucia|L|', 'Das|Dividutta|D|', 'Wang|Xinhong|X|', 'Luo|Haibei|H|', 'Zungu-Edmondson|Makhosazane|M|', 'Suzuki|Yuichiro J|YJ|']
D000447:Aldehydes D000818:Animals D002478:Cells, Cultured D004137:Dinitrochlorobenzene D054477:Glutaredoxins D006801:Humans D008297:Male D008315:Malondialdehyde D008623:Mercaptoethanol D018384:Oxidative Stress D054464:Peroxiredoxins D010659:Phenylhydrazines D050050:Protein Carbonylation D034622:RNA Interference D034741:RNA, Small Interfering D051381:Rats D017207:Rats, Sprague-Dawley D017382:Reactive Oxygen Species D013438:Sulfhydryl Compounds D013879:Thioredoxins
Decarbonylation Carbonylation 2,4-dinitrophenylhydrazine Peroxiredoxin Peroxiredoxin 2,4-dinitrochlorobenzene 2,4-dinitrophenyl BME BSA DNCB DNP DNPH DTT ECL Free radicals GSH Grx1 HRP IEF N-acetyl-l-cysteine NAC Oxidant signaling Oxidative stress PDGF Protein oxidation Prx ROS Reactive oxygen species Redox signaling SDS SMC Trx bovine serum albumin dithiothreitol enhanced chemiluminescence glutaredoxin-1 horseradish peroxidase isoelectric focusing l-glutathione peroxiredoxin platelet-derived growth factor reactive oxygen species smooth muscle cells sodium dodecyl sulfate thioredoxin β-mercaptoethanol