Kobayashi S, Rikans L E
Comp Biochem Physiol B. 1984;77(2):313-8. doi: 10.1016/0305-0491(84)90335-3.
NADPH-cytochrome P-450 reductase was purified to apparent homogeneity from detergent-solubilized guinea pig liver microsomes. The reductase had a mol. wt of 78,000 and contained one mole each of FAD and FMN. Electron transfer activity to cytochrome c was optimal at a pH of 8.0 and an ionic strength of 0.43. The results of kinetic experiments were consistent with a ternary-complex mechanism for the interaction of the reductase with cytochrome c and NADPH. Km values for NADPH and cytochrome c were 3.1 and 26.7 microM, respectively. Inhibition by NADP+ and 2'-AMP was competitive with respect to NADPH; Ki values were 12.1 microM for NADP+ and 46.7 microM for 2'-AMP.
从去污剂增溶的豚鼠肝脏微粒体中纯化出了具有明显均一性的NADPH-细胞色素P-450还原酶。该还原酶的分子量为78,000，含有一摩尔的黄素腺嘌呤二核苷酸（FAD）和一摩尔的黄素单核苷酸（FMN）。向细胞色素c的电子转移活性在pH值为8.0和离子强度为0.43时最佳。动力学实验结果与还原酶与细胞色素c和NADPH相互作用的三元复合物机制一致。NADPH和细胞色素c的米氏常数（Km）分别为3.1微摩尔和26.7微摩尔。NADP⁺和2'-AMP的抑制作用对NADPH而言是竞争性的；NADP⁺的抑制常数（Ki）为12.1微摩尔，2'-AMP的抑制常数为46.7微摩尔。
