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High-throughput fluorescence assay of cytochrome P450 3A4.

Cheng Q, Guengerich FP
Methods Mol Biol. 2013 987: 157-62

PMID: 23475676 · PMCID: PMC4024206 · DOI:10.1007/978-1-62703-321-3_14

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.

MeSH Terms (8)

Coumarins Cytochrome P-450 CYP3A Cytochrome P-450 CYP3A Inhibitors Enzyme Assays Humans Inhibitory Concentration 50 Ketoconazole Spectrometry, Fluorescence

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