Abstract
Cytochrome P450 2E1 was isolated from minipig liver microsomes. The protein has been cloned and the respective cDNA sequenced (GenBank Accession Number AY581116). Minipig CYP2E1 is two residues shorter than its human ortholog. The only difference between pig and minipig sequence is the presence of aspartic acid residue in position 346 contrary to valine in the pig enzyme. Minipig CYP2E1 was shown to be able to convert two prototypical substrates of human CYP2E1, chlorzoxazone and p-nitrophenol, to the respective metabolites. The experiments performed with both the liver microsomal fraction and reconstituted systems with human or minipig CYP2E1 confirmed the similarity of both enzymes. Inhibition with diethyldithiocarbamate gave comparable Ki values for minipig as well as for the human CYP2E1. The results indicate that the systems containing minipig CYP2E1 may be used to model the respective CYP2E1-catalyzed reactions of drug metabolism in humans.
Footnotes
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Financial support from the Grant Agency of the Czech Republic (Project No 203/02/1152) is gratefully acknowledged.
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Part of the results was presented at the 13th International Conference, Cytochromes P450, Biochemistry, Biophysics and Drug Metabolism, Prague, Czech Republic, June 29-July 3, 2003.
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Article, publication date, and citation information can be found at http://dmd.aspetjournals.org.
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doi:10.1124/dmd.104.003392.
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ABBREVIATIONS: P450, cytochrome P450; PCR, polymerase chain reaction.
- Received December 17, 2004.
- Accepted March 16, 2005.
- The American Society for Pharmacology and Experimental Therapeutics
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