title: Liver microsomes from the yellow rat snake (Elaphe obsoleta) and American bullfrog (Rana catesbeiana) oxidize polyunsaturated fatty acids by NADPH-dependent hydroxylation and epoxidation.
abstract: Polyunsaturated fatty acids (PUFAs) can be oxygenated by mammalian hepatic P450s to a series of metabolites. The most prominent of these are formed by omega- and (omega-1)-hydroxylation, epoxidation of the double bonds or bisallylic hydroxylation. The object of the present investigation was to determine whether similar oxygenations are catalyzed by liver microsomes of the yellow rat snake (Elaphe obsoleta) and the American bullfrog (Rana catesbeiana). Liver microsomes were incubated with [1-14C]-labeled arachidonic (AA), eicosapentaenoic (EPA), and linoleic acids (LA) in the presence or absence of 1 mM NADPH, and the major metabolites were analyzed by reverse-phase and straight-phase high performance liquid chromatography and capillary gas chromatography-mass spectroscopy. No metabolites were produced in the absence of NADPH. Profiles of metabolites were different depending on the organism and the acclimation state. In all incubations, EPA was the most effective substrate utilized and LA the least effective. The major products from EPA were 19-HEPE, 13-HEPE, and 20-HEPE from cold-acclimated (5 degrees C), warm-acclimated (22 degrees C) frogs, and snakes (22 degrees C), respectively. In contrast, 20-HETE production from AA was greater than 19-HETE in all three. Cold-acclimated frog liver microsomes produced significantly more of all metabolites when compared with microsomes from warm-acclimated frogs. We conclude that amphibian and snake liver can catalyze epoxidation and hydroxylation of PUFAs and that products are species-specific and acclimation-state dependent.
authors.affiliation: Department of Biology, New Mexico State University, Las Cruces 88003, USA. cherman@nmsu.edu
authors.forename: C A
authors.initials: CA
authors.lastname: Herman
authors.forename: E H
authors.initials: EH
authors.lastname: Oliw
mesh_terms: D000064:Acclimatization / Q000502:physiology; D000818:Animals; D016718:Arachidonic Acid / Q000378:metabolism; D002250:Carbon Radioisotopes; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003577:Cytochrome P-450 Enzyme System / Q000032:analysis; D015118:Eicosapentaenoic Acid / Q000378:metabolism; D005231:Fatty Acids, Unsaturated / Q000378:metabolism*; D008401:Gas Chromatography-Mass Spectrometry; D006893:Hydroxyeicosatetraenoic Acids / Q000378:metabolism; D006900:Hydroxylation; D019787:Linoleic Acid / Q000378:metabolism; D008099:Liver / Q000378:metabolism / Q000648:ultrastructure; D008862:Microsomes, Liver / Q000378:metabolism* / Q000648:ultrastructure; D019255:NADPH Oxidases / Q000502:physiology*; D010084:Oxidation-Reduction; D011892:Rana catesbeiana / Q000378:metabolism*; D012911:Snakes / Q000378:metabolism*; D013045:Species Specificity
chemical_list: D002250:Carbon Radioisotopes; D005231:Fatty Acids, Unsaturated; D006893:Hydroxyeicosatetraenoic Acids; D016718:Arachidonic Acid; D003577:Cytochrome P-450 Enzyme System; D019787:Linoleic Acid; D015118:Eicosapentaenoic Acid; D019255:NADPH Oxidases