title: Identification of the human liver cytochrome P-450 responsible for coumarin 7-hydroxylase activity.
abstract: 1. We have constructed a full-length human liver cytochrome P450IIA cDNA from a partial-length clone by oligonucleotide-directed mutagenesis, and subcloned it into the monkey kidney (COS-7) cell expression vector, pSVL. 2. The cDNA encodes a 49 kDa protein with coumarin 7-hydroxylase (COH) activity which cross-reacts with antisera to the mouse cytochrome P-450 isoenzyme responsible for COH activity and comigrates with a human liver microsomal protein. 3. Western blot analysis of a panel of human livers indicates that the level of the 49 kDa protein, detected using antisera to either the mouse COH P-450 or rat P450IIA1 protein, correlates very highly with COH activity. 4. Antisera to the rat P450IIA1 protein can inhibit COH activity in human liver microsomes. Taken together, these data indicate that a member of the P450IIA subfamily is responsible for most, if not all, of the COH activity in human liver.
authors.affiliation: Imperial Cancer Research Fund, University Department of Biochemistry, George Square, Edinburgh, U.K.
authors.forename: J S
authors.initials: JS
authors.lastname: Miles
authors.forename: A W
authors.initials: AW
authors.lastname: McLaren
authors.forename: L M
authors.initials: LM
authors.lastname: Forrester
authors.forename: M J
authors.initials: MJ
authors.lastname: Glancey
authors.forename: M A
authors.initials: MA
authors.lastname: Lang
authors.forename: C R
authors.initials: CR
authors.lastname: Wolf
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001189:Aryl Hydrocarbon Hydroxylases; D001483:Base Sequence; D002460:Cell Line; D065637:Cytochrome P-450 CYP2A6; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics* / Q000378:metabolism; D004247:DNA / Q000235:genetics; D006801:Humans; D008099:Liver / Q000201:enzymology*; D006899:Mixed Function Oxygenases / Q000235:genetics* / Q000378:metabolism*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009154:Mutation; D015345:Oligonucleotide Probes; D012689:Sequence Homology, Nucleic Acid; D014162:Transfection
chemical_list: D015345:Oligonucleotide Probes; D004247:DNA; D003577:Cytochrome P-450 Enzyme System; D006899:Mixed Function Oxygenases; D001189:Aryl Hydrocarbon Hydroxylases; D065637:Cytochrome P-450 CYP2A6