Bovine luteal cytochrome P-450scc was purified and incorporated into artificial phosphatidylcholine vesicles. The vesicle reconstituted cytochrome used membrane bound cholesterol as substrate and cholesterol binding varied with the phosphatidylcholine fatty acyl composition and was stimulated by the presence of cardiolipin in the vesicle. 22R -hydroxycholesterol and 20 alpha, 22R - dihydroxycholesterol bound to the cytochrome up to 300 times more tightly than cholesterol and decreased the affinity of the cytochrome for CO by 100-200-fold. The properties of the cytochrome closely paralleled those reported for cytochrome P-450scc purified from the bovine adrenal gland.