title: High-throughput fluorescence assay of cytochrome P450 3A4.
abstract: Microtiter plate-based fluorescence assays allow rapid measurement of the catalytic activities of cytochrome P450 oxygenases (P450s). We describe a high-throughput fluorescence assay of P450 3A4, one of the key enzymes involved in xenobiotic metabolism. The assay involves the oxidative debenzylation of 7-hydroxy-4-trifluoromethyl coumarin, producing an increase in fluorescence.
authors.affiliation: Department of Biochemistry and Center in Molecular Toxicology, Vanderbilt University School of Medicine, Nashville, TN, USA.
authors.forename: Qian
authors.initials: Q
authors.lastname: Cheng
authors.forename: F Peter
authors.initials: FP
authors.lastname: Guengerich
mesh_terms: D003374:Coumarins / Q000737:chemistry / Q000378:metabolism; D051544:Cytochrome P-450 CYP3A / Q000378:metabolism*; D065692:Cytochrome P-450 CYP3A Inhibitors; D057075:Enzyme Assays / Q000379:methods*; D006801:Humans; D020128:Inhibitory Concentration 50; D007654:Ketoconazole / Q000494:pharmacology; D013050:Spectrometry, Fluorescence / Q000379:methods*
chemical_list: D003374:Coumarins; D065692:Cytochrome P-450 CYP3A Inhibitors; C030123:coumarin; D051544:Cytochrome P-450 CYP3A; D007654:Ketoconazole