title: Cytochromes P450 catalyze the reduction of α,β-unsaturated aldehydes.
abstract: The metabolism of α,β-unsaturated aldehydes, e.g., 4-hydroxynonenal, involves oxidation to carboxylic acids, reduction to alcohols, and glutathionylation to eventually form mercapturide conjugates. Recently, we demonstrated that P450s can oxidize aldehydes to carboxylic acids, a reaction previously thought to involve aldehyde dehydrogenase. When recombinant cytochrome P450 3A4 was incubated with 4-hydroxynonenal, O(2), and NADPH, several products were produced, including 1,4-dihydroxynonene (DHN), 4-hydroxy-2-nonenoic acid (HNA), and an unknown metabolite. Several P450s catalyzed the reduction reaction in the order (human) P450 2B6 ≅ P450 3A4 > P450 1A2 > P450 2J2 > (mouse) P450 2c29. Other P450s did not catalyze the reduction reaction (human P450 2E1 and rabbit P450 2B4). Metabolism by isolated rat hepatocytes showed that HNA formation was inhibited by cyanamide, while DHN formation was not affected. Troleandomycin increased HNA production 1.6-fold while inhibiting DHN formation, suggesting that P450 3A11 is a major enzyme involved in rat hepatic clearance of 4-HNE. A fluorescent assay was developed using 9-anthracenealdehyde to measure both reactions. Feeding mice a diet containing t-butylated hydroxyanisole increased the level of both activities with hepatic microsomal fractions but not proportionally. Miconazole (0.5 mM) was a potent inhibitor of these microsomal reduction reactions, while phenytoin and α-naphthoflavone (both at 0.5 mM) were partial inhibitors, suggesting the role of multiple P450 enzymes. The oxidative metabolism of these aldehydes was inhibited >90% in an Ar or CO atmosphere, while the reductive reactions were not greatly affected. These results suggest that P450s are significant catalysts of the reduction of α,β-unsaturated aldehydes in the liver.
authors.affiliation: Department of Biochemistry and Molecular Biology, The University of Louisville School of Medicine , Louisville, KY 40292, USA.
authors.forename: Immaculate
authors.initials: I
authors.lastname: Amunom
authors.forename: Laura J
authors.initials: LJ
authors.lastname: Dieter
authors.forename: Viola
authors.initials: V
authors.lastname: Tamasi
authors.forename: Jian
authors.initials: J
authors.lastname: Cai
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Conklin
authors.forename: Sanjay
authors.initials: S
authors.lastname: Srivastava
authors.forename: Martha V
authors.initials: MV
authors.lastname: Martin
authors.forename: F Peter
authors.initials: FP
authors.lastname: Guengerich
authors.forename: Russell A
authors.initials: RA
authors.lastname: Prough
mesh_terms: D000447:Aldehydes / Q000737:chemistry / Q000378:metabolism*; D000818:Animals; D000873:Anthracenes / Q000737:chemistry / Q000378:metabolism; D055162:Biocatalysis; D002478:Cells, Cultured; D003484:Cyanamide / Q000494:pharmacology; D051544:Cytochrome P-450 CYP3A / Q000235:genetics / Q000378:metabolism*; D022781:Hepatocytes / Q000187:drug effects; D006801:Humans; D006880:Hydroxy Acids / Q000737:chemistry / Q000378:metabolism; D008297:Male; D051379:Mice; D008862:Microsomes, Liver / Q000378:metabolism; D009249:NADP / Q000378:metabolism; D010084:Oxidation-Reduction; D010100:Oxygen / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014217:Troleandomycin / Q000494:pharmacology
chemical_list: C091766:4-hydroxy-2-nonenoic acid; D000447:Aldehydes; D000873:Anthracenes; D006880:Hydroxy Acids; D011994:Recombinant Proteins; D003484:Cyanamide; D009249:NADP; C073050:9-anthraldehyde; D014217:Troleandomycin; D051544:Cytochrome P-450 CYP3A; C027576:4-hydroxy-2-nonenal; D010100:Oxygen