title: Luminogenic cytochrome P450 assays.
abstract: Luminogenic cytochrome P450 (CYP) assays couple CYP enzyme activity to firefly luciferase luminescence in a technology called P450-Glo(TM) (Promega). Luminogenic substrates are used in assays of human CYP1A1, -1A2, -1B1, -2C8, -2C9, -2C19, -2D6, -2J2, -3A4, -3A7, -4A11, -4F3B, -4F12 and -19. The assays detect dose-dependent CYP inhibition by test compounds against recombinant CYP enzymes or liver microsomes. Induction or inhibition of CYP activities in cultured hepatocytes is measured in a nonlytic approach that leaves cells intact for additional analysis. Luminogenic CYP assays offer advantages of speed and safety over HPLC and radiochemical-based methods. Compared with fluorogenic methods the approach offers advantages of improved sensitivity and decreased interference between optical properties of test compound and CYP substrate. These homogenous assays are sensitive and robust tools for high-throughput CYP screening in early drug discovery.
authors.affiliation: Promega Corp., Madison, WI 53711, USA. jim.cali@promega.com
authors.forename: James J
authors.initials: JJ
authors.lastname: Cali
authors.forename: Dongping
authors.initials: D
authors.lastname: Ma
authors.forename: Mary
authors.initials: M
authors.lastname: Sobol
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Simpson
authors.forename: Susan
authors.initials: S
authors.lastname: Frackman
authors.forename: Troy D
authors.initials: TD
authors.lastname: Good
authors.forename: William J
authors.initials: WJ
authors.lastname: Daily
authors.forename: David
authors.initials: D
authors.lastname: Liu
mesh_terms: D000465:Algorithms; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D065607:Cytochrome P-450 Enzyme Inhibitors; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D004791:Enzyme Inhibitors; D005456:Fluorescent Dyes; D006801:Humans; D008156:Luciferases / Q000378:metabolism; D049408:Luminescent Agents / Q000737:chemistry*; D013379:Substrate Specificity
chemical_list: D065607:Cytochrome P-450 Enzyme Inhibitors; D004791:Enzyme Inhibitors; D005456:Fluorescent Dyes; D049408:Luminescent Agents; D003577:Cytochrome P-450 Enzyme System; D008156:Luciferases