title: Stimulation of adrenal mitochondrial cholesterol side-chain cleavage by GTP, steroidogenesis activator polypeptide (SAP), and sterol carrier protein2. GTP and SAP act synergistically.
abstract: Several factors are known to stimulate cholesterol side-chain cleavage in isolated adrenal mitochondria, including steroidogenesis activator polypeptide (SAP), GTP, and sterol carrier protein2 (SCP2). All of these reportedly function at the level of the translocation of cholesterol to the inner membrane wherein side-chain cleavage to form pregnenolone occurs. We have investigated the activating effects of these factors alone and in combination. Under conditions where exogenous cholesterol is provided and multiple turnovers of a transport system are required, GTP stimulated steroidogenesis in isolated mitochondria and in adrenal homogenates, and this effect was enhanced by a GTP regenerating system. SAP alone had little effect under these conditions, but synergized with GTP to stimulate cholesterol metabolism. A truncated SAP analog and a variant from the C terminus of the minor heat-shock protein GRP78 had similar effects, but an unrelated peptide had no effect. GTP stimulated side-chain cleavage with the same EC50 in both resting mitochondria (from dexamethasone-treated rats) and in activated mitochondria (from ether-treated rats), but SAP effects were most apparent in resting mitochondria. In contrast, SCP2 stimulation was additive with other factors, suggesting an independent mechanism of action. While the data are consistent with biological roles for these factors, the relatively small magnitude of the in vitro effects may indicate that cell disruption and mitochondrial isolation disrupt important structural or other features which are necessary for the full expression of the steroidogenic response.
authors.affiliation: Department of Biochemistry, Emory University School of Medicine, Atlanta, Georgia 30322.
authors.forename: T S
authors.initials: TS
authors.lastname: Xu
authors.forename: E P
authors.initials: EP
authors.lastname: Bowman
authors.forename: D B
authors.initials: DB
authors.lastname: Glass
authors.forename: J D
authors.initials: JD
authors.lastname: Lambeth
mesh_terms: D000311:Adrenal Glands / Q000201:enzymology*; D000595:Amino Acid Sequence; D000818:Animals; D002352:Carrier Proteins / Q000494:pharmacology*; D002786:Cholesterol Side-Chain Cleavage Enzyme / Q000378:metabolism*; D003907:Dexamethasone / Q000494:pharmacology; D004357:Drug Synergism; D004986:Ether / Q000494:pharmacology; D005260:Female; D006160:Guanosine Triphosphate / Q000378:metabolism / Q000494:pharmacology*; D006360:Heat-Shock Proteins; D007700:Kinetics; D008928:Mitochondria / Q000201:enzymology*; D018832:Molecular Chaperones; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000494:pharmacology; D010940:Plant Proteins; D011284:Pregnenolone / Q000096:biosynthesis; D011506:Proteins / Q000494:pharmacology*; D051381:Rats; D011919:Rats, Inbred Strains; D000091342:Endoplasmic Reticulum Chaperone BiP
chemical_list: D002352:Carrier Proteins; D002786:Cholesterol Side-Chain Cleavage Enzyme; D003907:Dexamethasone; D004986:Ether; D006160:Guanosine Triphosphate; D006360:Heat-Shock Proteins; D018832:Molecular Chaperones; D010446:Peptide Fragments; D010940:Plant Proteins; D011284:Pregnenolone; D011506:Proteins; C051684:GRP78 protein, rat; D000091342:Endoplasmic Reticulum Chaperone BiP; C072087:sterol carrier proteins