Luminogenic cytochrome P450 assays.
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.
Cali
James J
JJ
Promega Corp., Madison, WI 53711, USA. jim.cali@promega.com
Ma
Dongping
D
Sobol
Mary
M
Simpson
Daniel J
DJ
Frackman
Susan
S
Good
Troy D
TD
Daily
William J
WJ
Liu
David
D
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
D065607:Cytochrome P-450 Enzyme Inhibitors; D004791:Enzyme Inhibitors; D005456:Fluorescent Dyes; D049408:Luminescent Agents; D003577:Cytochrome P-450 Enzyme System; D008156:Luciferases