Purification of adrenodoxin reductase from bovine adrenal cortex mitochondria by affinity chromatography. Properties of steroid hydroxylase systems reconstituted from adrenodoxin reductase, adrenodoxin and membranous cytochrome P-45011β

Adrenodoxin reductase from bovine adrenal cortex mitochondria was purified to homogeneity by affinity chromatography on 2',5'-ADP-Sepharose 4B. The purified adrenodoxin reductase was reconstitutively active in the presence of NADPH and adrenodoxin and with either cytochrome c or cytochrome P-450 as electron acceptor. In sonicated cytochrome P-45-containing milochondrial membranes reconstituted with adrenodoxin reductase and adrenodoxin. the content of cytochrome P-450 and level of reduction of cytochrome P-450 was not altered as compared to intact mitochondria. In contrast, hydroxylation of deoxycorticosteron was inhibited, suggesting that a component of cytochrome P-450 or an hypothetical factor required for hydroxylation of steroids by membrane-bound cytochrome P-450, was impaired.