High-throughput fluorescence assay of cytochrome P450 3A4

被引:13
作者
Cheng, Qian
Sohl, Christal D.
Guengerich, F. Peter [1 ]
机构
[1] Vanderbilt Univ, Sch Med, Dept Biochem, Nashville, TN 37212 USA
关键词
DRUG-DRUG INTERACTIONS; INHIBITION; P450; POINT;
D O I
10.1038/nprot.2009.123
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Cytochrome P450 mono-oxygenases (P450s) are the principal enzymes involved in the oxidative metabolism of drugs and other xenobiotics. In this protocol, we describe a fluorescence-based, high-throughput assay for measuring the activity of P450 3A4, one of the key enzymes involved in drug metabolism. The assay involves the oxidative debenzylation of a substituted coumarin, yielding an increase in fluorescence on reaction. The entire procedure can be accomplished in 1 h or less.
引用
收藏
页码:1258 / 1261
页数:4
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