Purification and characterization of 25-hydroxyvitamin D3 1alpha-hydroxylase from rat kidney mitochondria.
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.
Nakamura
Y
Y
Department of Surgery I, Miyazaki Medical College, Kiyotake, Japan.
Eto
T A
TA
Taniguchi
T
T
Miyamoto
K
K
Nagatomo
J
J
Shiotsuki
H
H
Sueta
H
H
Higashi
S
S
Okuda
K I
KI
Setoguchi
T
T
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
D003577:Cytochrome P-450 Enzyme System; D015090:25-Hydroxyvitamin D3 1-alpha-Hydroxylase