Stable Expression of Human Cytochrome P450 3A4 in Conjunction with Human NADPH-Cytochrome P450 Oxidoreductase in V79 Chinese Hamster Cells
| Autor: | Wolfgang A. Schmalix, D Lang, G.J. Horbach, R Bocker, P Beaune, C Belloc, Helmut Greim, E.M. de Groene, V Siruguri, A Schneider, B Kleingeist, Johannes Doehmer |
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| Rok vydání: | 1996 |
| Předmět: |
Aflatoxin B1
DNA Complementary CYP2B6 Genetic Vectors 25-Hydroxyvitamin D3 1-alpha-hydroxylase Biophysics Gene Expression Biology Reductase Transfection Biochemistry Cell Line Mixed Function Oxygenases Cytochrome P-450 Enzyme System Cricetinae Animals Cytochrome P-450 CYP3A Humans Molecular Biology NADPH-Ferrihemoprotein Reductase Micronucleus Tests CYP3A4 Cytochrome c CYP1A2 Cytochrome P450 reductase Recombinant Proteins Coenzyme Q – cytochrome c reductase biology.protein |
| Zdroj: | Archives of Biochemistry and Biophysics. 332:295-304 |
| ISSN: | 0003-9861 |
| DOI: | 10.1006/abbi.1996.0345 |
| Popis: | V79 Chinese hamster cells were constructed for stable expression of human cytochrome P450 3A4 with and without coexpression of human NADPH-cytochrome P450 oxidoreductase. Expression of the cDNAs was shown by Northern and Western analyses. Activity was tested by 6 beta-hydroxylation of testosterone for cytochrome P450 3A4 and by cytochrome c reduction for NADPH-cytochrome P450 reductase. Five V79 cell lines were obtained expressing cytochrome P450 3A4, human NADPH-cytochrome P450 oxidoreductase, and both. Cytochrome P450 3A4 activity depended highly on cytochrome P450 reductase activity, with lowest activity when only the parental Chinese hamster cytochrome P450 reductase was present, 5- and 10-fold higher when coexpressed with the human NADPH-cytochrome P450 reductase. Correspondingly, cytotoxic and genotoxic potency of aflatoxin B1 was increased by orders of magnitudes when human cytochrome P450 3A4 was coexpressed with the human NADPH-cytochrome P450 reductase. The effect of NADPH-cytochrome P450 reductase coexpression on cytochrome P450 3A4 activity was also tested by nifedipine oxidation and midazolam hydroxylation. Nifedipine oxidation was increased about 10-fold, 1-hydroxylation of midazolam and 4-hydroxylation of midazolam were increased 15-fold. |
| Databáze: | OpenAIRE |
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