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. 1983 Apr;80(7):1877–1881. doi: 10.1073/pnas.80.7.1877

Modulation of the kinetics of cholesterol side-chain cleavage by an activator and by an inhibitor isolated from the cytosol of the cortex of bovine adrenals.

P A Warne, N J Greenfield, S Lieberman
PMCID: PMC393713  PMID: 6572949

Abstract

Two modulators of sterol side-chain cleavage activity have been detected in the cytosol from the cortex of bovine adrenals. One is an inhibitor of side-chain cleavage which increases the Km of a purified and reconstituted mitochondrial side-chain cleavage system for both cholesterol and cholesterol sulfate. It also lowers the Vmax of cleavage when cholesterol sulfate is the substrate. The other modulator is a low molecular weight protein which in the reconstituted system increases the Vmax of cleavage for both substrates but does not affect the Km of either. Side-chain cleavage activity was also found in bovine adrenocortical cytosol and this appears to differ from that present in mitochondria.

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Selected References

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

  1. CALVIN H. I., LIEBERMAN S. EVIDENCE THAT STEROID SULFATES SERVE AS BIOSYNTHETIC INTERMEDIATES. II. IN VITRO CONVERSION OF PREGNENOLONE-3H SULFATE-35S TO 17ALPHA-HYDROXYPREGNENOLONE-3H SULFATE-35S. Biochemistry. 1964 Feb;3:259–264. doi: 10.1021/bi00890a020. [DOI] [PubMed] [Google Scholar]
  2. Davis W. W., Garren L. D. On the mechanism of action of adrenocorticotropic hormone. The inhibitory site of cycloheximide in the pathway of steroid biosynthesis. J Biol Chem. 1968 Oct 10;243(19):5153–5157. [PubMed] [Google Scholar]
  3. DuBois R. N., Simpson E. R., Kramer R. E., Waterman M. R. Induction of synthesis of cholesterol side chain cleavage cytochrome P-450 by adrenocorticotropin in cultured bovine adrenocortical cells. J Biol Chem. 1981 Jul 10;256(13):7000–7005. [PubMed] [Google Scholar]
  4. Erickson S. K., Meyer D. J., Gould R. G. Purification and characterization of a new cholesterol-binding protein from rat liver cytosol. J Biol Chem. 1978 Mar 25;253(6):1817–1826. [PubMed] [Google Scholar]
  5. 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]
  6. GOA J. A micro biuret method for protein determination; determination of total protein in cerebrospinal fluid. Scand J Clin Lab Invest. 1953;5(3):218–222. doi: 10.3109/00365515309094189. [DOI] [PubMed] [Google Scholar]
  7. Gasparini F., Wolfson A., Hochberg R., Lieberman S. Side chain cleavage of some cholesterol esters. J Biol Chem. 1979 Jul 25;254(14):6650–6656. [PubMed] [Google Scholar]
  8. Greenfield N. J., Gerolimatos B., Szwergold B. S., Wolfson A. J., Prasad V. V., Lieberman S. Effects of phospholipid and detergent on the substrate specificity of adrenal cytochrome P-450scc. Substrate binding and kinetics of cholesterol side chain oxidation. J Biol Chem. 1981 May 10;256(9):4407–4417. [PubMed] [Google Scholar]
  9. Hall P. F., Watanuki M., Hamkalo B. A. Adrenocortical cytochrome P-450 side chain cleavage. Preparation of membrane-bound side chain cleavage system from purified components. J Biol Chem. 1979 Jan 25;254(2):547–552. [PubMed] [Google Scholar]
  10. Hanukoglu I., Jefcoate C. R. Mitochondrial cytochrome P-450scc. Mechanism of electron transport by adrenodoxin. J Biol Chem. 1980 Apr 10;255(7):3057–3061. doi: 10.1016/S0021-9258(19)85851-9. [DOI] [PubMed] [Google Scholar]
  11. Hochberg R. B., Hoeven TA vander, Welch M., Lieberman S. A simple and precise assay of the enzymatic conversion of cholesterol into pregnenolone. Biochemistry. 1974 Jan 29;13(3):603–609. doi: 10.1021/bi00700a031. [DOI] [PubMed] [Google Scholar]
  12. Kan K. W., Ungar F. Characterization of an adrenal activator for cholesterol side chain cleavage. J Biol Chem. 1973 Apr 25;248(8):2868–2875. [PubMed] [Google Scholar]
  13. Katagiri M., Takemori S., Itagaki E., Suhara K. Purification of adrenal cytochrome P-450 (cholesterol desmolase and steroid 11beta- and 18-hydroxylase). Methods Enzymol. 1978;52:124–132. doi: 10.1016/s0076-6879(78)52014-4. [DOI] [PubMed] [Google Scholar]
  14. Koritz S. B., Kumar A. M. On the mechanism of action of the adrenocorticotrophic hormone. The stimulation of the activity of enzymes involved in pregnenolone synthesis. J Biol Chem. 1970 Jan 10;245(1):152–159. [PubMed] [Google Scholar]
  15. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  16. Lambeth J. D., Kamin H. Adrenodoxin reductase.adrenodoxin complex. Flavin to iron-sulfur electron transfer as the rate-limiting step in the NADPH-cytochrome c reductase reaction. J Biol Chem. 1979 Apr 25;254(8):2766–2774. [PubMed] [Google Scholar]
  17. Lambeth J. D., Seybert D. W., Kamin H. Phospholipid vesicle-reconstituted cytochrome P-450SCC. Mutually facilitated binding of cholesterol and adrenodoxin. J Biol Chem. 1980 Jan 10;255(1):138–143. [PubMed] [Google Scholar]
  18. Lefevre A., Morera A. M., Saez J. M. Adrenal cholesterol-binding protein: properties and partial purification. FEBS Lett. 1978 May 15;89(2):287–292. doi: 10.1016/0014-5793(78)80238-5. [DOI] [PubMed] [Google Scholar]
  19. Mellon-Nussbaum S., Welch M., Bandy L., Lieberman S. The lipoidal derivatives of steroids as biosynthetic intermediates. J Biol Chem. 1980 Mar 25;255(6):2487–2492. [PubMed] [Google Scholar]
  20. Merril C. R., Goldman D., Sedman S. A., Ebert M. H. Ultrasensitive stain for proteins in polyacrylamide gels shows regional variation in cerebrospinal fluid proteins. Science. 1981 Mar 27;211(4489):1437–1438. doi: 10.1126/science.6162199. [DOI] [PubMed] [Google Scholar]
  21. Nakajin S., Ishii Y., Shinoda M., Shikita M. Binding of Triton X-100 to purified cytochrome P-450scc and enhancement of the cholesterol side chain cleavage activity. Biochem Biophys Res Commun. 1979 Mar 30;87(2):524–531. doi: 10.1016/0006-291x(79)91827-8. [DOI] [PubMed] [Google Scholar]
  22. ROBERTS K. D., BANDI L., CALVIN H. I., DRUCKER W. D., LIEBERMAN S. EVIDENCE THAT STEROID SULFATES SERVE AS BIOSYNTHETIC INTERMEDIATES. IV. CONVERSION OF CHOLESTEROL SULFATE IN VIVO TO URINARY C-19 AND C-21 STEROIDAL SULFATES. Biochemistry. 1964 Dec;3:1983–1988. doi: 10.1021/bi00900a034. [DOI] [PubMed] [Google Scholar]
  23. Ritter M. C., Dempsey M. E. Specificity and role in cholesterol biosynthesis of a squalene and sterol carrier protein. J Biol Chem. 1971 Mar 10;246(5):1536–1539. [PubMed] [Google Scholar]
  24. Strott C. A., Lyons C. D. Two classes of cholesterol binding activity in the soluble fraction of the adrenal cortex of the guinea pig. J Steroid Biochem. 1980 Jan;13(1):73–82. doi: 10.1016/0022-4731(80)90115-6. [DOI] [PubMed] [Google Scholar]
  25. Suhara K., Takemori S., Katagiri M. Improved purification of bovine adrenal iron-sulfur protein. Biochim Biophys Acta. 1972 Apr 15;263(2):272–278. doi: 10.1016/0005-2795(72)90079-7. [DOI] [PubMed] [Google Scholar]
  26. Teicher B. A., Shikita M., Talalay P. Effects of adrenal steroid activator protein on the conversion of various 20- and 22-hydroxycholesterols to pregnenolone by adrenal mitochondrial enzymes. Biochem Biophys Res Commun. 1978 Aug 29;83(4):1436–1441. doi: 10.1016/0006-291x(78)91381-5. [DOI] [PubMed] [Google Scholar]
  27. Wolfson A. J., Lieberman S. Evidence suggesting that more than one sterol side chain cleavage enzyme system exists in mitochondria from bovine adrenal cortex. J Biol Chem. 1979 May 25;254(10):4094–4100. [PubMed] [Google Scholar]

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