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 ass...

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Published inExpert opinion on drug metabolism & toxicology Vol. 2; no. 4; p. 629
Main Authors Cali, James J, Ma, Dongping, Sobol, Mary, Simpson, Daniel J, Frackman, Susan, Good, Troy D, Daily, William J, Liu, David
Format Journal Article
LanguageEnglish
Published England 01.08.2006
Subjects
Algorithms
Animals
Chromatography, High Pressure Liquid
Cytochrome P-450 Enzyme Inhibitors
Cytochrome P-450 Enzyme System - metabolism
Enzyme Inhibitors
Fluorescent Dyes
Humans
Luciferases - metabolism
Luminescent Agents - chemistry
Substrate Specificity
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Summary: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.
ISSN:1742-5255
DOI:10.1517/17425255.2.4.629