title: Assay of Endocannabinoid Oxidation by Cytochrome P450.
abstract: Cytochrome P450 enzymes are a large family of heme-containing proteins that have important functions in the biotransformation of xenobiotics, including pharmacologic and environmental agents, as well as of endogenously produced chemicals with broad structural and functional diversity. Anandamide and 2-arachidonoylglycerol (2-AG) are substrates for P450s expressed in multiple tissues, leading to the production of a diverse set of mono- and di-oxygenated metabolites. This chapter describes tools and methods that have been used to identify major endocannabinoid-metabolizing P450s and their corresponding products, by using subcellular tissue fractions, cultured cells, and purified recombinant enzymes in a reconstituted system.
authors.affiliation: Department of Cell Biology and Physiology, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA. ntsnider@med.unc.edu.
authors.forename: Natasha T
authors.initials: NT
authors.lastname: Snider
authors.affiliation: Department of Medicine - Nephrology, University of Alabama at Birmingham, Birmingham, AL, USA.
authors.forename: Vyvyca J
authors.initials: VJ
authors.lastname: Walker
authors.affiliation: Department of Pharmacology, University of Michigan, Ann Arbor, MI, USA.
authors.forename: Paul F
authors.initials: PF
authors.lastname: Hollenberg
mesh_terms: D000818:Animals; D001681:Biological Assay / Q000379:methods; D002853:Chromatography, Liquid; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D063388:Endocannabinoids / Q000378:metabolism*; D006801:Humans; D013058:Mass Spectrometry; D008861:Microsomes / Q000378:metabolism; D008928:Mitochondria / Q000378:metabolism; D010084:Oxidation-Reduction
chemical_list: D063388:Endocannabinoids; D003577:Cytochrome P-450 Enzyme System
keywords: Eicosanoid; Liquid chromatography-mass spectrometry; Metabolism; Microsomes; Oxidation; P450 epoxygenase; P450 hydroxylase