Abstract
1. Protein degradation in rat hepatocytes in stationary monolayer culture was measured as release of radioactive trichloroacetic acid-soluble material from intracellular proteins labelled with [3H]leucine. 2. Glucocorticoids, but not other steroids, stimulated protein breakdown in the hepatocyte monolayers. The effects observed were greater when the cells were preincubated with the hormones, indicating that the stimulation was not immediate. In addition, the stimulation by glucocorticoids persisted for up to 4 h after hormone removal. 3. Cycloheximide and the lysosomotropic agents leupeptin and ammonia effectively blocked glucocorticoid stimulation of protein degradation. 4. Insulin blocked dexamethasone stimulation when added at the same time as the steroid, but not when added 3 h later. 5. Stimulation of protein breakdown by dexamethasone was additive with that by glucagon or dibutyryl cyclic AMP, suggesting that its mechanism of action is different from that of the latter two agents. 6. Total activities of several lysosomal enzymes were unaffected under conditions where protein breakdown was stimulated by either glucagon or dexamethasone. 7. It is suggested that, whereas glucagon, dibutyryl cyclic AMP and insulin modulate protein breakdown in these cells via changes in autophagocytosis, the stimulation by glucocorticoids is exerted independently, perhaps by stimulating the synthesis of membrane proteins essential to the autophagic process.
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Selected References
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- Arstila A. U., Shelburne J. D., Trump B. F. Studies on cellular autophagocytosis. A histochemical study on sequential alterations of mitochondria in the glucagon-induced autophagic vacuoles of rat liver. Lab Invest. 1972 Sep;27(3):317–323. [PubMed] [Google Scholar]
- BEAUFAY H., VAN CAMPENHOUT E., DE DUVE C. Tissue fractionation studies. II. Influence of various hepatotoxic treatments on the state of some bound enzymes in rat liver. Biochem J. 1959 Dec;73:617–623. doi: 10.1042/bj0730617. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ballard F. J. Intracellular protein degradation. Essays Biochem. 1977;13:1–37. [PubMed] [Google Scholar]
- Bodansky O. Acid phosphatase. Adv Clin Chem. 1972;15:43–147. doi: 10.1016/s0065-2423(08)60159-6. [DOI] [PubMed] [Google Scholar]
- Bowness J. M., Carpenter N., Barry D. N. Increased subcellular hyaluronidase in the rat after hydrocortisone injection. Can J Biochem. 1971 Jan;49(1):12–18. doi: 10.1139/o71-003. [DOI] [PubMed] [Google Scholar]
- Carubelli R., Griffin M. J. Neuraminidase activity in hela cells: effect of hydrocortisone. Science. 1970 Dec 4;170(3962):1110–1112. doi: 10.1126/science.170.3962.1110. [DOI] [PubMed] [Google Scholar]
- Chertow B. S., Buchanan W. E., Mayron M. S., Schwartz T. B. Hormonal modification of mouse liver lysosomal protein metabolism by cortisone acetate. Endocrinology. 1973 Mar;92(3):722–727. doi: 10.1210/endo-92-3-722. [DOI] [PubMed] [Google Scholar]
- Ciechanover A., Heller H., Elias S., Haas A. L., Hershko A. ATP-dependent conjugation of reticulocyte proteins with the polypeptide required for protein degradation. Proc Natl Acad Sci U S A. 1980 Mar;77(3):1365–1368. doi: 10.1073/pnas.77.3.1365. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cristofalo V. J., Kabakjian J. Lysosomal enzymes and aging in vitro: subcellular enzyme distribution and effect of hydrocortisone on cell life-span. Mech Ageing Dev. 1975 Jan-Feb;4(1):19–28. doi: 10.1016/0047-6374(75)90004-4. [DOI] [PubMed] [Google Scholar]
- DeMartino G. N., Goldberg A. L. Identification and partial purification of an ATP-stimulated alkaline protease in rat liver. J Biol Chem. 1979 May 25;254(10):3712–3715. [PubMed] [Google Scholar]
- Etlinger J. D., Goldberg A. L. A soluble ATP-dependent proteolytic system responsible for the degradation of abnormal proteins in reticulocytes. Proc Natl Acad Sci U S A. 1977 Jan;74(1):54–58. doi: 10.1073/pnas.74.1.54. [DOI] [PMC free article] [PubMed] [Google Scholar]
- FEIGELSON P., FEIGELSON M. Studies on the mechanism of regulation by cortisone of the metabolism of liver purine and ribonucleic acid. J Biol Chem. 1963 Mar;238:1073–1077. [PubMed] [Google Scholar]
- GOODLAD G. A., MUNRO H. N. Diet and the action of cortisone on protein metabolism. Biochem J. 1959 Oct;73:343–348. doi: 10.1042/bj0730343. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gelehrter T. D., Emanuel J. R. Hormonal regulation of protein degradation in hepatoma cells in tissue culture. Endocrinology. 1974 Mar;94(3):676–684. doi: 10.1210/endo-94-3-676. [DOI] [PubMed] [Google Scholar]
- Goldberg A. L., Dice J. F. Intracellular protein degradation in mammalian and bacterial cells. Annu Rev Biochem. 1974;43(0):835–869. doi: 10.1146/annurev.bi.43.070174.004155. [DOI] [PubMed] [Google Scholar]
- Goldberg A. L. Protein turnover in skeletal muscle. II. Effects of denervation and cortisone on protein catabolism in skeletal muscle. J Biol Chem. 1969 Jun 25;244(12):3223–3229. [PubMed] [Google Scholar]
- Goldberg A. L., St John A. C. Intracellular protein degradation in mammalian and bacterial cells: Part 2. Annu Rev Biochem. 1976;45:747–803. doi: 10.1146/annurev.bi.45.070176.003531. [DOI] [PubMed] [Google Scholar]
- Grammeltvedt R., Berg T. Influence of insulin on lysosomal activity and urea production in isolated parenchymal cells from rat liver. Hoppe Seylers Z Physiol Chem. 1976 Jul;357(7):977–981. doi: 10.1515/bchm2.1976.357.2.977. [DOI] [PubMed] [Google Scholar]
- Gunn J. M., Ballard F. J., Hanson R. W. Infulence of hormones and medium composition on the degradation of phosphoenolpyruvate carboxykinase (GTP) and total protein in Reuber H35 cells. J Biol Chem. 1976 Jun 25;251(12):3586–3593. [PubMed] [Google Scholar]
- Hanoune J., Chambaut A. M., Josipowicz A. The glucose effect and cortisone action upon rat liver and muscle protein metabolism. Arch Biochem Biophys. 1972 Jan;148(1):180–184. doi: 10.1016/0003-9861(72)90129-4. [DOI] [PubMed] [Google Scholar]
- Hanoune J., Feigelson P. Turnover of protein and RNA of liver ribosomal components in normal and cortisol-treated rats. Biochim Biophys Acta. 1970 Jan 21;199(1):214–223. doi: 10.1016/0005-2787(70)90710-0. [DOI] [PubMed] [Google Scholar]
- Henderson I. C., Fischel R. E., Loeb J. N. Suppression of liver DNA synthesis by cortisone. Endocrinology. 1971 Jun;88(6):1471–1476. doi: 10.1210/endo-88-6-1471. [DOI] [PubMed] [Google Scholar]
- Hershko A., Ciechanover A., Heller H., Haas A. L., Rose I. A. Proposed role of ATP in protein breakdown: conjugation of protein with multiple chains of the polypeptide of ATP-dependent proteolysis. Proc Natl Acad Sci U S A. 1980 Apr;77(4):1783–1786. doi: 10.1073/pnas.77.4.1783. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hille M. B., Barrett A. J., Dingle J. T., Fell H. B. Microassay for cathepsin D shows an unexpected effedt of cycloheximide on limb-bone rudiments in organ culture. Exp Cell Res. 1970 Aug;61(2):470–472. doi: 10.1016/0014-4827(70)90476-3. [DOI] [PubMed] [Google Scholar]
- Hopgood M. F., Clark M. G., Ballard F. J. Inhibition of protein degradation in isolated rat hepatocytes. Biochem J. 1977 May 15;164(2):399–407. doi: 10.1042/bj1640399. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hopgood M. F., Clark M. G., Ballard F. J. Protein degradation in hepatocyte monolayers. Effects of glucagon, adenosine 3':5'-cyclic monophosphate and insulin. Biochem J. 1980 Jan 15;186(1):71–79. doi: 10.1042/bj1860071. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ikezawa H., Aoyagi T., Takeuchi T., Umezawa H. Effect of protease inhibitors of actinomycetes on lysosomal peptide-hydrolases from swine liver. J Antibiot (Tokyo) 1971 Jul;24(7):488–490. doi: 10.7164/antibiotics.24.488. [DOI] [PubMed] [Google Scholar]
- Kim Y. S., Kim Y. Glucocorticoid inhibition of protein synthesis in vivo and in vitro. J Biol Chem. 1975 Mar 25;250(6):2293–2298. [PubMed] [Google Scholar]
- Knowles S. E., Ballard F. J. Selective control of the degradation of normal and aberrant proteins in Reuber H35 hepatoma cells. Biochem J. 1976 Jun 15;156(3):609–617. doi: 10.1042/bj1560609. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Knowles S. E., Gunn J. M., Hanson R. W., Ballard F. J. Increased degradation rates of protein synthesized in hepatoma cells in the presence of amino acid analogues. Biochem J. 1975 Mar;146(3):595–600. doi: 10.1042/bj1460595. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kyaw A., Mellors A. The effect of cortisone acetate on lysosomal enzyme levels in rat liver. Can J Biochem. 1972 Jan;50(1):20–24. doi: 10.1139/o72-004. [DOI] [PubMed] [Google Scholar]
- L'Age M., Langholz J., Fechner W., Salzmann H. Disturbances of the hypothalamo-hypophysial-adrenocortical system in the alloxan diabetic rat. Endocrinology. 1974 Sep;95(3):760–765. doi: 10.1210/endo-95-3-760. [DOI] [PubMed] [Google Scholar]
- Lundgren E. Isozyme variations in human cells grown in vitro. 8. Glucocorticoid regulation of amino acid naphthylamidase. Hum Hered. 1972;22(1):45–54. doi: 10.1159/000152467. [DOI] [PubMed] [Google Scholar]
- Neely A. N., Cox J. R., Fortney J. A., Schworer C. M., Mortimore G. E. Alterations of lysosomal size and density during rat liver perfusion. Suppression by insulin and amino acids. J Biol Chem. 1977 Oct 10;252(19):6948–6954. [PubMed] [Google Scholar]
- Pfeifer U. Inhibited autophagic degradation of cytoplasm during compensatory growth of liver cells after partial hepatectomy. Virchows Arch B Cell Pathol Incl Mol Pathol. 1979 Jun 29;30(3):313–333. doi: 10.1007/BF02889111. [DOI] [PubMed] [Google Scholar]
- Pfeifer U. Inhibition by insulin of the physiological autophagic breakdown of cell organelles. Acta Biol Med Ger. 1977;36(11-12):1691–1694. [PubMed] [Google Scholar]
- Plattner H., Henning R. Isolation of hepatocyte-derived rat liver lysosomal fractions. Exp Cell Res. 1975 Mar 15;91(2):333–343. doi: 10.1016/0014-4827(75)90112-3. [DOI] [PubMed] [Google Scholar]
- Reijngoud D. J., Oud P. S., Kás J., Tager J. M. Relationship between medium pH and that of the lysosomal matrix as studied by two independent methods. Biochim Biophys Acta. 1976 Oct 5;448(2):290–302. doi: 10.1016/0005-2736(76)90243-1. [DOI] [PubMed] [Google Scholar]
- Rosa F. Ultrastructural changes produced by glucagon, cyclic 3'5'-AMP and epinephrine on perfused rat livers. J Ultrastruct Res. 1971 Feb;34(3):205–213. doi: 10.1016/s0022-5320(71)80068-0. [DOI] [PubMed] [Google Scholar]
- Rose I. A., Warms J. V., Hershko A. A high molecular weight protease in liver cytosol. J Biol Chem. 1979 Sep 10;254(17):8135–8138. [PubMed] [Google Scholar]
- Shelburne J. D., Arstila A. U., Trump B. F. Studies on cellular autophagocytosis. Cyclic AMP- and dibutyryl cyclic AMP-stimulated autophagy in rat liver. Am J Pathol. 1973 Sep;72(3):521–540. [PMC free article] [PubMed] [Google Scholar]
- Suzuki H., Osawa M., Endo H. Effect of hydrocortisone on HeLa cells with special reference to the RNA metabolism. Endocrinol Jpn. 1969 Feb;16(1):121–127. doi: 10.1507/endocrj1954.16.121. [DOI] [PubMed] [Google Scholar]
- Symons A. M., Lewis D. A., Ancill R. J. Stabilising action of anti-inflammatory steroids on lysosomes. Biochem Pharmacol. 1969 Oct;18(10):2581–2582. doi: 10.1016/0006-2952(69)90376-1. [DOI] [PubMed] [Google Scholar]
- Thompson E. B., Lippman M. E. Mechanism of action of glucocorticoids. Metabolism. 1974 Feb;23(2):159–202. doi: 10.1016/0026-0495(74)90113-9. [DOI] [PubMed] [Google Scholar]
- Tomas F. M., Munro H. N., Young V. R. Effect of glucocorticoid administration on the rate of muscle protein breakdown in vivo in rats, as measured by urinary excretion of N tau-methylhistidine. Biochem J. 1979 Jan 15;178(1):139–146. doi: 10.1042/bj1780139. [DOI] [PMC free article] [PubMed] [Google Scholar]
- WEISSMANN G. LABILIZATION AND STABILIZATION OF LYSOSOMES. Fed Proc. 1964 Sep-Oct;23:1038–1044. [PubMed] [Google Scholar]
- WEISSMANN G., THOMAS L. THE EFFECTS OF CORTICOSTEROIDS UPON CONNECTIVE TISSUE AND LYSOSOMES. Recent Prog Horm Res. 1964;20:215–245. [PubMed] [Google Scholar]
- WOOL I. G., WEINSHELBAUM E. I. Adrenal cortical hormone and incorporation of C14 from amino acid precursors into muscle protein. Am J Physiol. 1960 Feb;198:360–362. doi: 10.1152/ajplegacy.1960.198.2.360. [DOI] [PubMed] [Google Scholar]
- Warburton M. J., Poole B. Effect of medium composition on protein degradation and DNA synthesis in rat embryo fibroblasts. Proc Natl Acad Sci U S A. 1977 Jun;74(6):2427–2431. doi: 10.1073/pnas.74.6.2427. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wiener J., Loud A. V., Kimberg D. V., Spiro D. A quantitative description of cortisone-induced alterations in the ultrastructure of rat liver parenchmal cells. J Cell Biol. 1968 Apr;37(1):47–61. doi: 10.1083/jcb.37.1.47. [DOI] [PMC free article] [PubMed] [Google Scholar]
