Physical Incorporation of NADPH-Cytochrome P450 Reductase and Cytochrome P450 into Phospholipid Vesicles Using Glycocholate and Bio-Beads

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Physical incorporation of NADPH-cytochrome P450 reductase and cytochrome P450 into phospholipid vesicles using glycocholate and Bio-Beads.

In a previous study from our laboratory (Drug Metab Dispos 34: 660-666, 2006), we found several limitations with published methods (cholate gel filtration and cholate dialysis) for the incorporation of cytochromes P450 and P450 reductase into phospholipid vesicles. We found that a significant proportion of reductase was not incorporated in the vesicles when the amount of reductase was equal to ...

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We have incorporated CYP3A4 (cytochrome P450 3A4) and CPR (NADPH-cytochrome P450 reductase) into liposomes with a high lipid/protein ratio by an improved method. In the purified proteoliposomes, CYP3A4 binds testosterone with Kd (app)=36±6 μM and Hill coefficient=1.5±0.3, and 75±4% of the CYP3A4 can be reduced by NADPH in the presence of testosterone. Transfer of the first electron from CPR to ...

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ژورنال

عنوان ژورنال: Drug Metabolism and Disposition

سال: 2007

ISSN: 0090-9556,1521-009X

DOI: 10.1124/dmd.107.018473