Sono H, Sonoda Y, Sato Y
Kyoritsu College of Pharmacy, Tokyo, Japan.
Biochim Biophys Acta. 1991 Jul 12;1078(3):388-94. doi: 10.1016/0167-4838(91)90161-r.
Cytochrome P-45014DM, which catalyzes lanosterol 14 alpha-demethylation, from pig liver microsomes was purified to a state of virtually homogeneous by gel electrophoresis. Its apparent monomeric molecular weight was estimated to be 53,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and the amino-terminal amino acid sequence was Gly-Leu-Leu-Thr-Gly(Leu)-Asp-Leu-Leu-Gly-Ile. When reconstituted with NADPH-cytochrome P-450-reductase, the enzyme showed a high activity for lanosterol and 24,25-dihydrolanosterol 14 alpha-demethylation. Furthermore, the oxygenated intermediates of 24,25-dihydrolanosterol 14 alpha-demethylation, 32-hydroxy-24,25-dihydrolanosterol and 32-oxo-24,25-dihydrolanosterol, were converted to the 32-nor compound, 4,4-dimethylcholesta-8,14-dien-3 beta-ol, by the reconstituted enzyme system.
来自猪肝微粒体的催化羊毛甾醇14α-去甲基化的细胞色素P-45014DM通过凝胶电泳纯化至几乎均一的状态。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳估计其表观单体分子量为53,000，其氨基末端氨基酸序列为Gly-Leu-Leu-Thr-Gly(Leu)-Asp-Leu-Leu-Gly-Ile。当与NADPH-细胞色素P-450还原酶重组时，该酶对羊毛甾醇和24,25-二氢羊毛甾醇14α-去甲基化表现出高活性。此外，24,25-二氢羊毛甾醇14α-去甲基化的氧化中间体32-羟基-24,25-二氢羊毛甾醇和32-氧代-24,25-二氢羊毛甾醇通过重组酶系统转化为32-去甲化合物4,4-二甲基胆甾-8,14-二烯-3β-醇。
