Abstract
Adult bovine adrenal cortical cells in monolayer culture were used to study the induction of cholesterol side-chain-cleavage cytochrome P-450 by corticotropin (ACTH). In the presence of 1 microM ACTH, there was a 4-fold increase in cortisol production by these cells over a 72-hr period and a corresponding increase in total cytochrome P-450 content. The incorporation of [35S]methionine into a number of cellular proteins was stimulated by the presence of ACTH in the culture medium, whereas the incorporation into other proteins was decreased. The temporal profile of these changes varied from one protein to another. Examination of the incorporation of [35S]methionine into mitochondrial protein showed an increased production of a radiolabeled protein that comigrated with the form of cytochrome P-450 known as side-chain-cleavage cytochrome upon incubation with ACTH. Thus, it appears that the cytochrome P-450scc content is increased in bovine adrenal cortical cells exposed to ACTH. Cytochrome P-450scc, synthesized in a cell-free translation system directed by RNA isolated from bovine adrenal cortical tissue or from cells, had a molecular weight of 54,500. Cytochrome P-450scc isolated from bovine adrenal mitochondria had a molecular weight of 49,000. Thus, cytochrome P-450scc is synthesized as a larger precursor that must be processed by proteolytic cleavage before or upon insertion into the mitochondrion.
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- Asano K., Harding B. W. Biosynthesis of adrenodoxin in mouse adrenal tumor cells. Endocrinology. 1976 Oct;99(4):977–987. doi: 10.1210/endo-99-4-977. [DOI] [PubMed] [Google Scholar]
- 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]
- Bryson M. J., Sweat M. L. Cleavage of cholesterol side chain associated with cytochrome P-450, flavoprotein, and nonheme iron-protein derived from the bovine adrenal cortex. J Biol Chem. 1968 May 25;243(10):2799–2804. [PubMed] [Google Scholar]
- Côté C., Solioz M., Schatz G. Biogenesis of the cytochrome bc1 complex of yeast mitochondria. A precursor form of the cytoplasmically made subunit V. J Biol Chem. 1979 Mar 10;254(5):1437–1439. [PubMed] [Google Scholar]
- Dazord A., Gallet D., Saez J. M. Effect of corticotropin treatment in vivo on the synthesis of a specific adrenal cytosolic protein. Characterization by dual-labelling technique and polyacrylamide-gel electrophoresis. Biochem J. 1978 Oct 15;176(1):233–239. doi: 10.1042/bj1760233. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Deeley R. G., Gordon J. I., Burns A. T., Mullinix K. P., Binastein M., Goldberg R. F. Primary activation of the vitellogenin gene in the rooster. J Biol Chem. 1977 Nov 25;252(22):8310–8319. [PubMed] [Google Scholar]
- Gomez-Sanchez C., Milewich L., Holland O. B. Radioiodinated derivatives for steroid radioimmunoassay. Application to the radioimmunoassay of cortisol. J Lab Clin Med. 1977 Apr;89(4):902–909. [PubMed] [Google Scholar]
- Gospodarowicz D., Ill C. R., Hornsby P. J., Gill G. N. Control of bovine adrenal cortical cell proliferation by fibroblast growth factor. Lack of effect of epidermal growth factor. Endocrinology. 1977 Apr;100(4):1080–1089. doi: 10.1210/endo-100-4-1080. [DOI] [PubMed] [Google Scholar]
- Hornsby P. J., Gill G. N. Characterization of adult bovine adrenocortical cells throughout their life span in tissue culture. Endocrinology. 1978 Mar;102(3):926–936. doi: 10.1210/endo-102-3-926. [DOI] [PubMed] [Google Scholar]
- Hornsby P. J., Gill G. N. Hormonal control of adrenocortical cell proliferation. Desensitization to ACTH and interaction between ACTH and fibroblast growth factor in bovine adrenocortical cell cultures. J Clin Invest. 1977 Aug;60(2):342–352. doi: 10.1172/JCI108782. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ivarie R. D., Jones P. P. A rapid sensitive assay for specific protein synthesis in cells and in cell-free translations: use of Staphylococcus aureus as an adsorbent for immune complexes. Anal Biochem. 1979 Aug;97(1):24–35. doi: 10.1016/0003-2697(79)90322-1. [DOI] [PubMed] [Google Scholar]
- 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]
- Kowal J., Simpson E. R., Estabrook R. W. Adrenal cells in tissue culture. V. On the specificity of the stimulation of 11beta-hydroxylation by adrenocorticotropin. J Biol Chem. 1970 May 10;245(9):2438–2443. [PubMed] [Google Scholar]
- Kuno H., Kihara H. K. Simple microassay of protein with membrane filter. Nature. 1967 Aug 26;215(5104):974–975. doi: 10.1038/215974a0. [DOI] [PubMed] [Google Scholar]
- 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]
- Lewin A. S., Gregor I., Mason T. L., Nelson N., Schatz G. Cytoplasmically made subunits of yeast mitochondrial F1-ATPase and cytochrome c oxidase are synthesized as individual precursors, not as polyproteins. Proc Natl Acad Sci U S A. 1980 Jul;77(7):3998–4002. doi: 10.1073/pnas.77.7.3998. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Maccecchini M. L., Rudin Y., Blobel G., Schatz G. Import of proteins into mitochondria: precursor forms of the extramitochondrially made F1-ATPase subunits in yeast. Proc Natl Acad Sci U S A. 1979 Jan;76(1):343–347. doi: 10.1073/pnas.76.1.343. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Maccecchini M. L., Rudin Y., Schatz G. Transport of proteins across the mitochondrial outer membrane. A precursor form of the cytoplasmically made intermembrane enzyme cytochrome c peroxidase. J Biol Chem. 1979 Aug 25;254(16):7468–7471. [PubMed] [Google Scholar]
- Neuwelt E., Stumpf D., Austin J., Kohler P. A monospecific antibody to human sulfatase A. Preparation, characterization and significance. Biochim Biophys Acta. 1971 Apr 27;236(1):333–346. doi: 10.1016/0005-2795(71)90182-6. [DOI] [PubMed] [Google Scholar]
- 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]
- Palmiter R. D. Magnesium precipitation of ribonucleoprotein complexes. Expedient techniques for the isolation of undergraded polysomes and messenger ribonucleic acid. Biochemistry. 1974 Aug 13;13(17):3606–3615. doi: 10.1021/bi00714a032. [DOI] [PubMed] [Google Scholar]
- Paul D. P., Gallant S., Orme-Johnson N. R., Orme-Johnson W. H., Brownie A. C. Temperature dependence of cholesterol binding to cytochrome P-450scc of the rat adrenal. Effect of adrenocorticotropic hormone and cycloheximide. J Biol Chem. 1976 Nov 25;251(22):7120–7126. [PubMed] [Google Scholar]
- Pelham H. R., Jackson R. J. An efficient mRNA-dependent translation system from reticulocyte lysates. Eur J Biochem. 1976 Aug 1;67(1):247–256. doi: 10.1111/j.1432-1033.1976.tb10656.x. [DOI] [PubMed] [Google Scholar]
- Purvis J. L., Canick J. A., Mason J. I., Estabrook R. W., McCarthy J. L. Lifetime of adrenal cytochrome P-450 as influenced by ACTH. Ann N Y Acad Sci. 1973;212:319–343. doi: 10.1111/j.1749-6632.1973.tb47605.x. [DOI] [PubMed] [Google Scholar]
- Ray D. B., Horst I. A., Kowal J. Adrenocorticotropic hormone increases specific proteins of the mitochondrial fraction that are translated inside or outside this organelle in cultured adrenal tumor cells. Proc Natl Acad Sci U S A. 1980 Aug;77(8):4648–4652. doi: 10.1073/pnas.77.8.4648. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Richardson M. C., Schulster D. Corticosteroidogenesis in isolated adrenal cells: effect of adrenocorticotrophic hormone, adenosine 3',5'-monophosphate and 1-24 adrenocorticotrophic hormone diazotized to polyacrylamide. J Endocrinol. 1972 Oct;55(1):127–139. doi: 10.1677/joe.0.0550127. [DOI] [PubMed] [Google Scholar]
- 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]
- Sayers G., Beall R. J., Seelig S. Isolated adrenal cells: adrenocorticotropic hormone, calcium, steroidogenesis, and cyclic adenosine monophosphate. Science. 1972 Mar 10;175(4026):1131–1133. doi: 10.1126/science.175.4026.1131. [DOI] [PubMed] [Google Scholar]
- Schmelzer E., Heinrich P. C. Synthesis of a larger precursor for the subunit IV of rat liver cytochrome c oxidase in a cell-free wheat germ system. J Biol Chem. 1980 Aug 25;255(16):7503–7506. [PubMed] [Google Scholar]
- Seybert D. W., Lancaster J. R., Jr, Lambeth J. D., Kamin H. Participation of the membrane in the side chain cleavage of cholesterol. Reconstitution of cytochrome P-450scc into phospholipid vesicles. J Biol Chem. 1979 Dec 10;254(23):12088–12098. [PubMed] [Google Scholar]
- Shore G. C., Carignan P., Raymond Y. In vitro synthesis of a putative precursor to the mitochondrial enzyme, carbamyl phosphate synthetase. J Biol Chem. 1979 May 10;254(9):3141–3144. [PubMed] [Google Scholar]
- Simonian M. H., Gill G. N. Regulation of deoxyribonucleic acid synthesis in bovine adrenocortical cells in culture. Endocrinology. 1979 Mar;104(3):588–595. doi: 10.1210/endo-104-3-588. [DOI] [PubMed] [Google Scholar]
- Simpson E. R., Boyd G. S. Partial resolution of the mixed-function oxidase involved in the cholesterol side-chain cleavage reaction in bovine adrenal mitochondria. Biochem Biophys Res Commun. 1967 Sep 27;28(6):945–950. doi: 10.1016/0006-291x(67)90071-x. [DOI] [PubMed] [Google Scholar]
- Simpson E. R., Boyd G. S. The cholesterol side-chain cleavage system of bovine adrenal cortex. Eur J Biochem. 1967 Oct;2(3):275–285. doi: 10.1111/j.1432-1033.1967.tb00136.x. [DOI] [PubMed] [Google Scholar]
- Simpson E. R., Williams-Smith D. L. Electron paramagnetic resonance spectra of mitochondrial and microsomal cytochrome P-450 from the rat adrenal. Biochim Biophys Acta. 1976 Oct 13;449(1):59–71. doi: 10.1016/0005-2728(76)90007-4. [DOI] [PubMed] [Google Scholar]
- Takemori S., Sato H., Gomi T., Suhara K., Katagiri M. Purification and properties of cytochrome P-450(11beta) from adrenocortical mitochondria. Biochem Biophys Res Commun. 1975 Dec 1;67(3):1151–1157. doi: 10.1016/0006-291x(75)90794-9. [DOI] [PubMed] [Google Scholar]
- Yago N., Kobayashi S., Sekiyama S., Kurokawa H., Iwai Y. Further studies on the submitochondrial localization of cholesterol side chain-cleaving enzyme system in hog adrenal cortex by sonic treatment. J Biochem. 1970 Dec;68(6):775–783. doi: 10.1093/oxfordjournals.jbchem.a129414. [DOI] [PubMed] [Google Scholar]