title: Eicosanoid formation by a cytochrome P450 isoform expressed in the pharynx of Caenorhabditis elegans.
abstract: Caenorhabditis elegans harbours several CYP (cytochrome P450) genes that are homologous with mammalian CYP isoforms important to the production of physiologically active AA (arachidonic acid) metabolites. We tested the hypothesis that mammals and C. elegans may share similar basic mechanisms of CYP-dependent eicosanoid formation and action. We focused on CYP33E2, an isoform related to the human AA-epoxygenases CYP2C8 and CYP2J2. Co-expression of CYP33E2 with the human NADPH-CYP reductase in insect cells resulted in the reconstitution of an active microsomal mono-oxygenase system that metabolized EPA (eicosapentaenoic acid) and, with lower activity, also AA to specific sets of regioisomeric epoxy- and hydroxy-derivatives. The main products included 17,18-epoxyeicosatetraenoic acid from EPA and 19-hydroxyeicosatetraenoic acid from AA. Using nematode worms carrying a pCYP33E2::GFP reporter construct, we found that CYP33E2 is exclusively expressed in the pharynx, where it is predominantly localized in the marginal cells. RNAi (RNA interference)-mediated CYP33E2 expression silencing as well as treatments with inhibitors of mammalian AA-metabolizing CYP enzymes, significantly reduced the pharyngeal pumping frequency of adult C. elegans. These results demonstrate that EPA and AA are efficient CYP33E2 substrates and suggest that CYP-eicosanoids, influencing in mammals the contractility of cardiomyocytes and vascular smooth muscle cells, may function in C. elegans as regulators of the pharyngeal pumping activity.
authors.affiliation: Max Delbrück Center for Molecular Medicine, Robert-Rössle-Strasse 10, 13125 Berlin, Germany.
authors.forename: Mandy
authors.initials: M
authors.lastname: Kosel
authors.forename: Waltraud
authors.initials: W
authors.lastname: Wild
authors.forename: Alexandra
authors.initials: A
authors.lastname: Bell
authors.forename: Michael
authors.initials: M
authors.lastname: Rothe
authors.forename: Carsten
authors.initials: C
authors.lastname: Lindschau
authors.forename: Christian E W
authors.initials: CE
authors.lastname: Steinberg
authors.forename: Wolf-Hagen
authors.initials: WH
authors.lastname: Schunck
authors.forename: Ralph
authors.initials: R
authors.lastname: Menzel
mesh_terms: D000818:Animals; D017173:Caenorhabditis elegans / Q000201:enzymology* / Q000378:metabolism; D029742:Caenorhabditis elegans Proteins / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular; D065607:Cytochrome P-450 Enzyme Inhibitors; D003577:Cytochrome P-450 Enzyme System / Q000145:classification* / Q000378:metabolism*; D015777:Eicosanoids / Q000235:genetics / Q000378:metabolism*; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology*; D020868:Gene Silencing; D009154:Mutation; D010614:Pharynx / Q000201:enzymology; D020033:Protein Isoforms
chemical_list: D029742:Caenorhabditis elegans Proteins; D065607:Cytochrome P-450 Enzyme Inhibitors; D015777:Eicosanoids; D020033:Protein Isoforms; D003577:Cytochrome P-450 Enzyme System