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. 1980 Jan 15;186(1):145–152. doi: 10.1042/bj1860145

Non-esterified cholesterol-rich adrenal lipid fractions. Preparation, properties and preferential utilization for cholesterol side-chain cleavage by corticotropin-stimulated adrenal mitochondria.

R V Farese, W J Prudente, L T Chuang
PMCID: PMC1161513  PMID: 6245638

Abstract

Rat adrenal 105,000 g supernatant contains two lipid moieties, 'lipid-I' and 'lipid-II' which contain non-esterified cholesterol and stimulate cholesterol side-chain cleavage in soluble or mitochondrial enzyme systems. Lipid-I contains relatively large low-density heat-stable particles, whereas lipid-II particles are smaller, more dense and heat-labile. Lipid-I and lipid-II can be separated from clear cytosol by ultracentrifugation and gel filtration respectively. Corticotropin plus cycloheximide treatment increases the non-esterified cholesterol concentrations in the lipid fractions, and stimulatory effects of lipids on cholesterol side-chain cleavage appear to correlate with non-esterified cholesterol concentrations therein. On addition of saturating amounts of cholesterol-rich lipid, pregnenolone synthesis and cholesterol binding to cytochrome P-450 are stimulated more in mitochondria from corticotropin-stimulated adrenals than in mitochondria from control or corticotropin-plus cycloheximide-stimulated adrenals. These results support the contention that the corticotropin-induced increase in mitochondrial cholesterol side-chain cleavage involves an increase in cholesterol utilization as well as an increase in cholesterol availability.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bell J. J., Harding B. W. The acute action of adrenocorticotropic hormone on adrenal steroidogenesis. Biochim Biophys Acta. 1974 May 29;348(2):285–298. doi: 10.1016/0005-2760(74)90240-9. [DOI] [PubMed] [Google Scholar]
  2. Billiar R. B., Eik-Nes K. B. Biosynthesis of corticoids in guinea pig adrenal slices the effect of ACTH on [I-14C]acetate incorporation into adrenal steroids. Biochim Biophys Acta. 1965 Jul 8;104(2):503–514. doi: 10.1016/0304-4165(65)90356-9. [DOI] [PubMed] [Google Scholar]
  3. Boyd G. S., Trzeciak W. H. Cholesterol metabolism in the adrenal cortex: studies on the mode of action of ACTH. Ann N Y Acad Sci. 1973;212:361–377. doi: 10.1111/j.1749-6632.1973.tb47607.x. [DOI] [PubMed] [Google Scholar]
  4. Brownie A. C., Alfano J., Jefcoate C. R., Orme-Johnson W., Beinert H., Simpson E. R. Effect of ACTH on adrenal mitochondrial cytochrome P-450 in the rat. Ann N Y Acad Sci. 1973;212:344–360. doi: 10.1111/j.1749-6632.1973.tb47606.x. [DOI] [PubMed] [Google Scholar]
  5. Davis W. W., Garren L. D. Evidence for the stimulation by adenocorticotropic hormone of the conversion of cholesterol esters to cholesterol in the adrenal, in vivo. Biochem Biophys Res Commun. 1966 Sep 8;24(5):805–810. doi: 10.1016/0006-291x(66)90398-6. [DOI] [PubMed] [Google Scholar]
  6. FARESE R. V. INHIBITION OF THE STEROIDOGENIC EFFECT OF ACTH AND INCORPORATION OF AMINO ACID INTO RAT ADRENAL PROTEIN IN VITRO BY CHLORAMPHENICOL. Biochim Biophys Acta. 1964 Aug 12;87:699–701. doi: 10.1016/0926-6550(64)90292-0. [DOI] [PubMed] [Google Scholar]
  7. FERGUSON J. J., Jr PROTEIN SYNTHESIS AND ADRENOCORTICOTROPIN RESPONSIVENESS. J Biol Chem. 1963 Aug;238:2754–2759. [PubMed] [Google Scholar]
  8. Farese R. V., Prudente W. J. Corticotropin-induced changes in a soluble desmolase preparation. Biochim Biophys Acta. 1978 Mar 1;539(2):142–161. doi: 10.1016/0304-4165(78)90002-8. [DOI] [PubMed] [Google Scholar]
  9. Farese R. V., Prudente W. J. On the requirement for protein synthesis during corticotropin-induced stimulation of cholesterol side chain cleavage in rat adrenal mitochondrial and solubilized desmolase preparations. Biochim Biophys Acta. 1977 Feb 28;496(2):567–570. doi: 10.1016/0304-4165(77)90339-7. [DOI] [PubMed] [Google Scholar]
  10. Farese R. V., Prudente W. J. On the role of calcium in adrenocorticotropin-induced changes in mitochondrial pregnenolone synthesis. Endocrinology. 1978 Oct;103(4):1264–1271. doi: 10.1210/endo-103-4-1264. [DOI] [PubMed] [Google Scholar]
  11. Farese R. V., Prudente W. J. On the role of intra-adrenal unesterified cholesterol in the steroidogenic effect of corticotropin. Biochim Biophys Acta. 1978 Nov 15;544(1):77–84. doi: 10.1016/0304-4165(78)90211-8. [DOI] [PubMed] [Google Scholar]
  12. Farese R. V. Stimulation of pregnenolone synthesis by ACTH in rat adrenal sections. Endocrinology. 1971 Oct;89(4):958–962. doi: 10.1210/endo-89-4-958. [DOI] [PubMed] [Google Scholar]
  13. Garren L. D., Gill G. N., Masui H., Walton G. M. On the mechanism of action of ACTH. Recent Prog Horm Res. 1971;27:433–478. doi: 10.1016/b978-0-12-571127-2.50035-3. [DOI] [PubMed] [Google Scholar]
  14. Garren L. D., Ney R. L., Davis W. W. Studies on the role of protein synthesis in the regulation of corticosterone production by adrenocorticotropic hormone in vivo. Proc Natl Acad Sci U S A. 1965 Jun;53(6):1443–1450. doi: 10.1073/pnas.53.6.1443. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. HARDING B. W., WONG S. H., NELSON D. H. CARBON MONOXIDE-COMBINING SUBSTANCES IN RAT ADRENAL. Biochim Biophys Acta. 1964 Nov 22;92:415–417. doi: 10.1016/0926-6569(64)90206-8. [DOI] [PubMed] [Google Scholar]
  16. Jefcoate C. R., Hume R., Boyd G. S. Separation of two forms of cytochrome P450 adrenal cortex mitochondria. FEBS Lett. 1970 Jul 15;9(1):41–44. doi: 10.1016/0014-5793(70)80306-4. [DOI] [PubMed] [Google Scholar]
  17. Leier D. J., Jungmann R. A. Adrenocorticotropic hormone and dibutryl adenosine cyclic monophosphate-mediated Ca2+ uptake by rat adrenal glands. Biochim Biophys Acta. 1973 Dec 5;329(2):196–210. doi: 10.1016/0304-4165(73)90284-5. [DOI] [PubMed] [Google Scholar]
  18. Mahaffee D., Reitz R. C., Ney R. L. The mechanism of action of adrenocroticotropic hormone. The role of mitochondrial cholesterol accumulation in the regulation of steroidogenesis. J Biol Chem. 1974 Jan 10;249(1):227–233. [PubMed] [Google Scholar]
  19. OMURA T., SATO R. THE CARBON MONOXIDE-BINDING PIGMENT OF LIVER MICROSOMES. I. EVIDENCE FOR ITS HEMOPROTEIN NATURE. J Biol Chem. 1964 Jul;239:2370–2378. [PubMed] [Google Scholar]
  20. Quarfordt S. H., Nathans A., Dowdee M., Hilderman H. L. Heterogeneity of human very low density lipoproteins by gel filtration chromatography. J Lipid Res. 1972 Jul;13(4):435–444. [PubMed] [Google Scholar]
  21. SILBER R. H., BUSCH R. D., OSLAPAS R. Practical procedure for estimation of corticosterone or hydrocortisone. Clin Chem. 1958 Aug;4(4):278–285. [PubMed] [Google Scholar]
  22. STONE D., HECHTER O. Studies on ACTH action in perfused bovine adrenals: the site of action of ACTH in corticosteroidogenesis. Arch Biochem Biophys. 1954 Aug;51(2):457–469. doi: 10.1016/0003-9861(54)90501-9. [DOI] [PubMed] [Google Scholar]
  23. Sata T., Estrich D. L., Wood P. D., Kinsell L. W. Evaluation of gel chromatography for plasma lipoprotein fractionation. J Lipid Res. 1970 Jul;11(4):331–340. [PubMed] [Google Scholar]
  24. Simpson E. R., Jefcoate C. R., Brownie A. C., Boyd G. S. The effect of ether anaesthesia stress on cholesterol-side-chain cleavage and cytochrome P450 in rat-adrenal mitochondria. Eur J Biochem. 1972 Jul 24;28(3):442–450. doi: 10.1111/j.1432-1033.1972.tb01931.x. [DOI] [PubMed] [Google Scholar]
  25. Simpson E. R., McCarthy J. L., Peterson J. A. Evidence that the cycloheximide-sensitive site of adrenocorticotropic hormone action is in the mitochondrion. Changes in pregnenolone formation, cholesterol content, and the electron paramagnetic resonance spectra of cytochrome P-450. J Biol Chem. 1978 May 10;253(9):3135–3139. [PubMed] [Google Scholar]

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