Regulation of cholesterol movement to mitochondrial cytochrome P450scc in steroid hormone synthesis.

Transfer of cholesterol to cytochrome P450scc is generally the rate-limiting step in steroid synthesis. Depending on the steroidogenic cell, cholesterol is supplied from low or high density lipoproteins (LDL or HDL) or de novo synthesis. ACTH and gonadotropins stimulate this cholesterol transfer prior to activation of gene transcription, both through increasing the availability of cytosolic free cholesterol and through enhanced cholesterol transfer between the outer and inner mitochondrial membranes. Cytosolic free cholesterol from LDL or HDL is primarily increased through enhanced cholesterol ester hydrolysis and suppressed esterification, but increased de novo synthesis can be significant. Elements of the cytoskeleton, probably in conjunction with sterol carrier protein(2) (SCP(2)), mediate cholesterol transfer to the mitochondrial outer membranes. Several factors contribute to the transfer of cholesterol between mitochondrial membranes; steroidogenesis activator peptide acts synergistically with GTP and is supplemented by SCP(2). 5-Hydroperoxyeicosatrienoic acid, endozepine (at peripheral benzodiazepine receptors), and rapid changes in outer membrane phospholipid content may also contribute stimulatory effects at this step. It is suggested that hormonal activation, through these factors, alters membrane structure around mitochondrial intermembrane contact sites, which also function to transfer ADP, phospholipids, and proteins to the inner mitochondria. Cholesterol transfer may occur following a labile fusion of inner and outer membranes, stimulated through involvement of cardiolipin and phosphatidylethanolamine in hexagonal phase membrane domains. Ligand binding to benzodiazepine receptors and the mitochondrial uptake of 37 kDa phosphoproteins that uniquely characterize steroidogenic mitochondria could possibly facilitate these changes. ACTH activation of rat adrenals increases the susceptibility of mitochondrial outer membranes to digitonin solubilization, suggesting increased cholesterol availability. Proteins associated with contact sites were not solubilized, indicating that this part of the outer membrane is resistant to this treatment. Two pools of reactive cholesterol within adrenal mitochondria have been distinguished by different isocitrate- and succinate-supported metabolism. These pools appear to be differentially affected in vitro by the above stimulatory factors.