title: Mechanism of protein decarbonylation.
abstract: 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.
authors.affiliation: Department of Pharmacology and Physiology, Georgetown University Medical Center, Washington, DC 20057, USA.
authors.forename: Chi-Ming
authors.initials: CM
authors.lastname: Wong
authors.affiliation: Department of Biochemical Sciences "A. Rossi Fanelli," Sapienza University of Rome, Rome, Italy.
authors.forename: Lucia
authors.initials: L
authors.lastname: Marcocci
authors.affiliation: Department of Pharmacology and Physiology, Georgetown University Medical Center, Washington, DC 20057, USA.
authors.forename: Dividutta
authors.initials: D
authors.lastname: Das
authors.affiliation: Department of Pharmacology and Physiology, Georgetown University Medical Center, Washington, DC 20057, USA; Department of Physiology and Pathophysiology, Fudan University Shanghai Medical College, Shanghai 200032, China.
authors.forename: Xinhong
authors.initials: X
authors.lastname: Wang
authors.affiliation: Department of Pharmacology and Physiology, Georgetown University Medical Center, Washington, DC 20057, USA.
authors.forename: Haibei
authors.initials: H
authors.lastname: Luo
authors.affiliation: Department of Pharmacology and Physiology, Georgetown University Medical Center, Washington, DC 20057, USA.
authors.forename: Makhosazane
authors.initials: M
authors.lastname: Zungu-Edmondson
authors.affiliation: Department of Pharmacology and Physiology, Georgetown University Medical Center, Washington, DC 20057, USA. Electronic address: ys82@georgetown.edu.
authors.forename: Yuichiro J
authors.initials: YJ
authors.lastname: Suzuki
mesh_terms: D000447:Aldehydes / Q000737:chemistry; D000818:Animals; D002478:Cells, Cultured; D004137:Dinitrochlorobenzene / Q000737:chemistry; D054477:Glutaredoxins / Q000737:chemistry* / Q000235:genetics; D006801:Humans; D008297:Male; D008315:Malondialdehyde / Q000737:chemistry; D008623:Mercaptoethanol / Q000737:chemistry; D018384:Oxidative Stress; D054464:Peroxiredoxins / Q000737:chemistry*; D010659:Phenylhydrazines / Q000737:chemistry; D050050:Protein Carbonylation / Q000502:physiology*; D034622:RNA Interference; D034741:RNA, Small Interfering; D051381:Rats; D017207:Rats, Sprague-Dawley; D017382:Reactive Oxygen Species / Q000737:chemistry*; D013438:Sulfhydryl Compounds / Q000737:chemistry; D013879:Thioredoxins / Q000235:genetics
chemical_list: D000447:Aldehydes; D004137:Dinitrochlorobenzene; D054477:Glutaredoxins; D010659:Phenylhydrazines; D034741:RNA, Small Interfering; D017382:Reactive Oxygen Species; D013438:Sulfhydryl Compounds; C004787:2,4-dinitrophenylhydrazine; D008315:Malondialdehyde; D013879:Thioredoxins; D008623:Mercaptoethanol; D054464:Peroxiredoxins; C027576:4-hydroxy-2-nonenal
keywords: 2,4-dinitrochlorobenzene; 2,4-dinitrophenyl; 2,4-dinitrophenylhydrazine; BME; BSA; Carbonylation; DNCB; DNP; DNPH; DTT; Decarbonylation; ECL; Free radicals; GSH; Grx1; HRP; IEF; N-acetyl-l-cysteine; NAC; Oxidant signaling; Oxidative stress; PDGF; Peroxiredoxin; 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
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