Utilization of intramitochondrial membrane cholesterol by cytochrome P-450-dependent cholesterol side-chain cleavage reaction in bovine adrenocortical mitochondria: steroidogenic and non-steroidogenic pools of cholesterol in the mitochondrial inner membranes

Abstract

Inner and outer submitochondrial membranes were prepared after disruption of malate-treated bovine adrenocortical mitochondria. It was found that a part of the endogenous cholesterol in the inner membrane (approx. 50%) was rapidly utilized by the cholesterol side-chain cleavage reaction. The utilization of cholesterol in the outer membrane, on the other hand, was inefficient and slow in spite of the fact that cholesterol concentration is higher in the outer than in the inner membrane. When the inner membrane prepared from untreated mitochondria was incubated for 20 min in the presence of a reconstituted cytochrome P-450-reducing system, the inner membrane cholesterol was depleted by approximately 70%. The half-life of the depletion reaction was 2-3 min. In addition, when the outer membrane plus the soluble fraction from the untreated mitochondria were added as a source of cholesterol to the inner membrane fraction, a marginal increase in the production of steroids was observed. From these results it is concluded that a portion of the inner membrane cholesterol can be steroidogenic, whereas the rest of the cholesterol is non-steroidogenic.