Synergistic stimulation of pregnenolone synthesis in rat adrenal mitochondria by n-hexane and cardiolipin.
n-Hexane and cardiolipin each stimulate pregnenolone production by isolated rat adrenal mitochondria. Following corticotropin (ACTH) stimulation, mitochondrial cholesterol metabolism exhibits a fast phase lasting 2 min, followed by a 10-fold slower metabolism. ACTH suppression by dexamethazone or cycloheximide (CX) treatment removes this fast phase. n-Hexane, at concentrations approaching 80% of the aqueous solubility limit (approximately 0.08 mM), selectively stimulates the slow phase of metabolism, while cardiolipin (100 microM) stimulates only the fast phase. Other alkanes and ethers are effective. The effect of n-hexane is dependent on mitochondrial integrity, as evidenced by decreased effects in hypoosmotically shocked mitochondria (outer membrane disrupted) and ineffectiveness in sonicated mitochondria (both membranes disrupted). n-Hexane apparently enhances the transfer of outer membrane cholesterol to inner membrane P-450scc. Stimulation by cardiolipin is retained by disrupted mitochondria and may involve enhanced availability of P-450scc to inner membrane cholesterol. When added together, these agents produce more than additive effects on cholesterol metabolism. Preincubation with n-hexane did not increase reactive cholesterol, suggesting that enhanced cholesterol transport occurs only in concert with metabolism of inner membrane cholesterol. Uptake of alkanes into mitochondrial membranes may effect structural changes that facilitate outer to inner membrane cholesterol transfer, but major changes are excluded by the effectiveness of isocitrate as a reductant for P-450scc. In combination, n-hexane and cardiolipin reproduce the effect of the ACTH-sensitive sterol regulatory peptide on mitochondria [R. C. Pedersen and A. C. Brownie (1983) Proc. Natl. Acad. Sci. USA 80, 1882-1886], suggesting that peptide action on adrenal mitochondria may resolve into two analogous components.
McNamara
B C
BC
Department of Pharmacology, University of Wisconsin Medical School, Madison 53706.
Jefcoate
C R
CR
D000311:Adrenal Glands / Q000187:drug effects / Q000378:metabolism*; D000324:Adrenocorticotropic Hormone / Q000494:pharmacology; D000616:Aminoglutethimide / Q000494:pharmacology; D000818:Animals; D002308:Cardiolipins / Q000494:pharmacology*; D002784:Cholesterol / Q000378:metabolism; D002786:Cholesterol Side-Chain Cleavage Enzyme / Q000378:metabolism; D003513:Cycloheximide / Q000494:pharmacology; D004357:Drug Synergism; D005260:Female; D006586:Hexanes / Q000494:pharmacology*; D007523:Isocitrates / Q000494:pharmacology; D007700:Kinetics; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D009249:NADP / Q000494:pharmacology; D011284:Pregnenolone / Q000096:biosynthesis*; D051381:Rats
D002308:Cardiolipins; D006586:Hexanes; D007523:Isocitrates; D000616:Aminoglutethimide; C026385:n-hexane; D009249:NADP; D011284:Pregnenolone; D000324:Adrenocorticotropic Hormone; D002784:Cholesterol; D003513:Cycloheximide; C034219:isocitric acid; D002786:Cholesterol Side-Chain Cleavage Enzyme