title: Purification and characterization of 25-hydroxyvitamin D3 1alpha-hydroxylase from rat kidney mitochondria.
abstract: We purified extensively 25-hydroxyvitamin D3 1alpha-hydroxylase (calcidiol, NADPH: oxygen oxidoreductase (1-hydroxylating), EC 1.14.13.13) from kidney mitochondria of rachitic rats and disclosed its peculiar properties as a P450. The final preparation was identified as a 55 kDa protein having an intense absorption at 417 nm characteristic of P450. The specific activity was 4.8 nmol/min/mg of protein indicating a 350-fold purification. Specific content of P450 was 1.1 nmol/mg of protein and turnover number was 4.4 min-1.
authors.affiliation: Department of Surgery I, Miyazaki Medical College, Kiyotake, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Nakamura
authors.forename: T A
authors.initials: TA
authors.lastname: Eto
authors.forename: T
authors.initials: T
authors.lastname: Taniguchi
authors.forename: K
authors.initials: K
authors.lastname: Miyamoto
authors.forename: J
authors.initials: J
authors.lastname: Nagatomo
authors.forename: H
authors.initials: H
authors.lastname: Shiotsuki
authors.forename: H
authors.initials: H
authors.lastname: Sueta
authors.forename: S
authors.initials: S
authors.lastname: Higashi
authors.forename: K I
authors.initials: KI
authors.lastname: Okuda
authors.forename: T
authors.initials: T
authors.lastname: Setoguchi
mesh_terms: D015090:25-Hydroxyvitamin D3 1-alpha-Hydroxylase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods; D003577:Cytochrome P-450 Enzyme System / Q000032:analysis; D004795:Enzyme Stability; D007668:Kidney / Q000201:enzymology*; D008297:Male; D008928:Mitochondria / Q000201:enzymology*; D008970:Molecular Weight; D051381:Rats; D017207:Rats, Sprague-Dawley; D012279:Rickets
chemical_list: D003577:Cytochrome P-450 Enzyme System; D015090:25-Hydroxyvitamin D3 1-alpha-Hydroxylase