title: Isolation of four forms of acetone-induced cytochrome P-450 in chicken liver by h.p.l.c. and their enzymic characterization.
abstract: The purpose of this study was to purify and characterize the forms of cytochrome P-450 induced in chicken liver by acetone or ethanol. Using high performance liquid ion-exchange chromatography, we were able to isolate at least four different forms of cytochrome P-450 which were induced by acetone in chicken liver. All four forms of cytochrome P-450 proved to be distinct proteins, as indicated by their N-terminal amino acid sequences and their reconstituted catalytic activities. Two of these forms, also induced by glutethimide in chicken embryo liver, appeared to be cytochromes P450IIH1 and P450IIH2. Both of these cytochromes P-450 have identical catalytic activities towards benzphetamine demethylation. However, they differ in their abilities to hydroxylate p-nitrophenol and to convert acetaminophen into a metabolite that forms a covalent adduct with glutathione at the 3-position. Another form of cytochrome P-450 induced by acetone is highly active in the hydroxylation of p-nitrophenol and in the conversion of acetaminophen to a reactive metabolite, similar to reactions catalysed by mammalian cytochrome P450IIE. Yet the N-terminal amino acid sequence of this form has only 30-33% similarity with cytochrome P450IIE purified from rat, rabbit and human livers. A fourth form of cytochrome P-450 was identified whose N-terminal amino acid sequence and enzymic activities do not correspond to any mammalian cytochromes P-450 reported to be induced by acetone or ethanol.
authors.affiliation: Veterans Administration, White River Junction, VT 05001.
authors.forename: J F
authors.initials: JF
authors.lastname: Sinclair
authors.forename: S
authors.initials: S
authors.lastname: Wood
authors.forename: L
authors.initials: L
authors.lastname: Lambrecht
authors.forename: N
authors.initials: N
authors.lastname: Gorman
authors.forename: L
authors.initials: L
authors.lastname: Mende-Mueller
authors.forename: L
authors.initials: L
authors.lastname: Smith
authors.forename: J
authors.initials: J
authors.lastname: Hunt
authors.forename: P
authors.initials: P
authors.lastname: Sinclair
mesh_terms: D000082:Acetaminophen / Q000378:metabolism; D000096:Acetone / Q000494:pharmacology*; D000595:Amino Acid Sequence; D000818:Animals; D001589:Benzphetamine / Q000378:metabolism; D002384:Catalysis; D002642:Chick Embryo; D002645:Chickens; D002851:Chromatography, High Pressure Liquid; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D000431:Ethanol / Q000494:pharmacology; D005984:Glutethimide / Q000494:pharmacology; D008099:Liver / Q000196:embryology / Q000201:enzymology*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009596:Nitrophenols / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D009596:Nitrophenols; D001589:Benzphetamine; D000096:Acetone; D000082:Acetaminophen; D000431:Ethanol; D003577:Cytochrome P-450 Enzyme System; D005984:Glutethimide; C024836:4-nitrophenol