Comparison of "high throughput" micromethods for determination of cytochrome P450 activities with classical methods using HPLC for product identification.
Enzyme activities of the CYP enzymes (CYP3A4, CYP2C9 and CYP2A6) were determined using classical substrates (testosterone, diclofenac and coumarin, respectively) as well as with luminogenic or fluorogenic substrates in micromethod arrangement. The luciferin-based luminogenic substrates for CYP3A4 and CYP2C9 as well as coumarin in micromethod for assay of CYP2A6 activity gave results well comparable with the classical methods with determination of reaction products by the HPLC.
Anzenbacherová
Eva
E
Institute of Medical Chemistry and Biochemistry, Palacky University, Hnevotínská 3, Olomouc, Czech Republic. anzeneva@tunw.upol.cz
Veinlichová
Alena
A
Masek
Vlastimil
V
Anzenbacher
Pavel
P
D001189:Aryl Hydrocarbon Hydroxylases / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid; D003374:Coumarins / Q000737:chemistry; D065637:Cytochrome P-450 CYP2A6; D065729:Cytochrome P-450 CYP2C9; D051544:Cytochrome P-450 CYP3A; D003577:Cytochrome P-450 Enzyme System / Q000032:analysis*; D004008:Diclofenac / Q000737:chemistry; D006801:Humans; D008297:Male; D006899:Mixed Function Oxygenases / Q000032:analysis*; D013739:Testosterone / Q000737:chemistry
D003374:Coumarins; D004008:Diclofenac; D013739:Testosterone; D003577:Cytochrome P-450 Enzyme System; C030123:coumarin; D006899:Mixed Function Oxygenases; C450260:CYP2C9 protein, human; D065729:Cytochrome P-450 CYP2C9; D001189:Aryl Hydrocarbon Hydroxylases; C585710:CYP2A6 protein, human; D065637:Cytochrome P-450 CYP2A6; D051544:Cytochrome P-450 CYP3A; C510163:CYP3A4 protein, human