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NADPH: cytochrome P450 reductases of various species can be used in systems reconstituting drug-metabolizing CYP2E1 activity
Marie Belejová, Eva Anzenbacherová, Roman Zuber, Pavel Anzenbacher
Jazyk angličtina Země Slovensko
- MeSH
- cytochrom P-450 CYP2E1 MeSH
- jaterní mikrozomy účinky léků MeSH
- krysa rodu rattus MeSH
- léčivé přípravky metabolismus MeSH
- miniaturní prasata MeSH
- NADP MeSH
- NADPH-cytochrom c-reduktasa MeSH
- systém (enzymů) cytochromů P-450 MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- zvířata MeSH
Metabolism of drugs and other foreign substances is mostly mediated by cytochromes P450 (P450, abbreviated also CYP for a particular enzyme). To find which P450 is involved in metabolism of a drug, liver microsomal monooxygenase system of P450 is reconstituted with its components: selected P450 enzyme, cytochrome b5, NADPH:P450 reductase and phospholipid. Used NADPH:P450 reductases were human recombinant and minipig or rat liver microsomal ones isolated by chromatographic separations. Chosen P450 enzyme was the CYP2E1 which is known to be of very similar primary structure among species; in this study, the minipig enzyme has been taken, as the minipigs seem to be suitable model animals for drug metabolism studies. Results obtained show that the reductase enzymes of rat and human origin can be used in reconstituted systems with a CYP2E1 as the activity of this enzyme varies in systems with reductases of different origin less than ten times. The results also indicate that the reductases from different species share a functional similarity, if not identity.
Literatura
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- $a Metabolism of drugs and other foreign substances is mostly mediated by cytochromes P450 (P450, abbreviated also CYP for a particular enzyme). To find which P450 is involved in metabolism of a drug, liver microsomal monooxygenase system of P450 is reconstituted with its components: selected P450 enzyme, cytochrome b5, NADPH:P450 reductase and phospholipid. Used NADPH:P450 reductases were human recombinant and minipig or rat liver microsomal ones isolated by chromatographic separations. Chosen P450 enzyme was the CYP2E1 which is known to be of very similar primary structure among species; in this study, the minipig enzyme has been taken, as the minipigs seem to be suitable model animals for drug metabolism studies. Results obtained show that the reductase enzymes of rat and human origin can be used in reconstituted systems with a CYP2E1 as the activity of this enzyme varies in systems with reductases of different origin less than ten times. The results also indicate that the reductases from different species share a functional similarity, if not identity.
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