title: NADP(+)-dependent dehydrogenase activity of carbonyl reductase on glutathionylhydroxynonanal as a new pathway for hydroxynonenal detoxification.
abstract: An NADP(+)-dependent dehydrogenase activity on 3-glutathionyl-4-hydroxynonanal (GSHNE) was purified to electrophoretic homogeneity from a line of human astrocytoma cells (ADF). Proteomic analysis identified this enzymatic activity as associated with carbonyl reductase 1 (EC 1.1.1.184). The enzyme is highly efficient at catalyzing the oxidation of GSHNE (KM 33 µM, kcat 405 min(-1)), as it is practically inactive toward trans-4-hydroxy-2-nonenal (HNE) and other HNE-adducted thiol-containing amino acid derivatives. Combined mass spectrometry and nuclear magnetic resonance spectroscopy analysis of the reaction products revealed that carbonyl reductase oxidizes the hydroxyl group of GSHNE in its hemiacetal form, with the formation of the corresponding 3-glutathionylnonanoic-δ-lactone. The relevance of this new reaction catalyzed by carbonyl reductase 1 is discussed in terms of HNE detoxification and the recovery of reducing power.
authors: Biochemistry Unit, Department of Biology, University of Pisa, I-56123 Pisa, Italy.
authors: Roberta
authors: R
authors: Moschini
authors: Biochemistry Unit, Department of Biology, University of Pisa, I-56123 Pisa, Italy.
authors: Eleonora
authors: E
authors: Peroni
authors: Biochemistry Unit, Department of Biology, University of Pisa, I-56123 Pisa, Italy.
authors: Rossella
authors: R
authors: Rotondo
authors: Proteomics & Mass Spectrometry Laboratory, ISPAAM-CNR, I-80147 Napoli, Italy.
authors: Giovanni
authors: G
authors: Renzone
authors: Institute of Biomolecular Chemistry, ICB-CNR, I-80078 Pozzuoli (Naples), Italy.
authors: Dominique
authors: D
authors: Melck
authors: Biochemistry Unit, Department of Biology, University of Pisa, I-56123 Pisa, Italy.
authors: Mario
authors: M
authors: Cappiello
authors: Health Unit 5 Pisa, Gynecology and Obstetric Unit, Pontedera Hospital, 56025 Pontedera, Italy.
authors: Massimo
authors: M
authors: Srebot
authors: Scuola Normale Superiore, Pisa, Italy.
authors: Elio
authors: E
authors: Napolitano
authors: Institute of Biomolecular Chemistry, ICB-CNR, I-80078 Pozzuoli (Naples), Italy.
authors: Andrea
authors: A
authors: Motta
authors: Proteomics & Mass Spectrometry Laboratory, ISPAAM-CNR, I-80147 Napoli, Italy.
authors: Andrea
authors: A
authors: Scaloni
authors: Biochemistry Unit, Department of Biology, University of Pisa, I-56123 Pisa, Italy.
authors: Umberto
authors: U
authors: Mura
authors: Biochemistry Unit, Department of Biology, University of Pisa, I-56123 Pisa, Italy. Electronic address: antonella.delcorso@unipi.it.
authors: Antonella
authors: A
authors: Del-Corso
mesh_terms: D000429:Alcohol Oxidoreductases / Q000302:isolation & purification / Q000378:metabolism*; D000449:Aldehyde Reductase / Q000378:metabolism; D000447:Aldehydes / Q000378:metabolism*; D001254:Astrocytoma / Q000378:metabolism* / Q000473:pathology; D005978:Glutathione / Q000031:analogs & derivatives* / Q000378:metabolism*; D006801:Humans; D008658:Inactivation, Metabolic; D007783:Lactones / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry; D009249:NADP / Q000378:metabolism*; D009252:NADPH Dehydrogenase / Q000378:metabolism*; D010084:Oxidation-Reduction; D040901:Proteomics; D013379:Substrate Specificity; D013438:Sulfhydryl Compounds / Q000378:metabolism; D014407:Tumor Cells, Cultured
chemical_list: C000605761:3-glutathionyl-4-hydroxynonanal; D000447:Aldehydes; D007783:Lactones; D013438:Sulfhydryl Compounds; D009249:NADP; D000429:Alcohol Oxidoreductases; C531129:CBR1 protein, human; D000449:Aldehyde Reductase; D009252:NADPH Dehydrogenase; D005978:Glutathione; C027576:4-hydroxy-2-nonenal
keywords: 3-Glutathionyl-4-hydroxynonanal; 4-Hydroxy-2-nonenal; Carbonyl reductase; Free radicals; Hydroxynonenal detoxification