Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1985 May 15;228(1):35–42. doi: 10.1042/bj2280035

Characterization of protein kinase C and its role in catecholamine secretion from bovine adrenal-medullary cells.

K W Brocklehurst, K Morita, H B Pollard
PMCID: PMC1144950  PMID: 3159383

Abstract

Protein kinase C activity towards exogenous histone was detected in a cytosolic fraction of bovine adrenal medulla. The enzyme was dependent on Ca2+ and phosphatidylserine for its activity, with half-maximal activation being achieved at approx. 18 microM free Ca2+ and 8 micrograms of phosphatidylserine/ml. Both diolein and 4 beta-phorbol 12-myristate 13-acetate (TPA) decreased the Ca2+ requirement of the enzyme, half-maximal activation being obtained at approx. 12 microM and 9 microM free Ca2+ respectively in the presence of these agents. Many endogenous proteins in the adrenal-medullary cytosolic fraction were detected whose phosphorylation was dependent on the presence of both Ca2+ and phosphatidylserine. TPA stimulated catecholamine release from cultured bovine adrenal-chromaffin cells in a Ca2+-dependent manner. A23187 also stimulated catecholamine secretion, and at sub-optimal concentrations of TPA and A23187 a synergistic secretory response was obtained. These results are consistent with protein kinase C having a regulatory role in exocytosis in bovine adrenal chromaffin cells.

Full text

PDF
35

Images in this article

39Fig. 6.
on p.39

Selected References

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

  1. Amy C. M., Kirshner N. Phosphorylation of adrenal medulla cell proteins in conjunction with stimulation of catecholamine secretion. J Neurochem. 1981 Mar;36(3):847–854. doi: 10.1111/j.1471-4159.1981.tb01671.x. [DOI] [PubMed] [Google Scholar]
  2. Berridge M. J. Inositol trisphosphate and diacylglycerol as second messengers. Biochem J. 1984 Jun 1;220(2):345–360. doi: 10.1042/bj2200345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  4. CREVELING C. R., DALY J. W., WITKOP B., UNDENFRIEND S. Substrates and inhibitors of dopamine-beta-oxidase. Biochim Biophys Acta. 1962 Oct 8;64:125–134. doi: 10.1016/0006-3002(62)90765-5. [DOI] [PubMed] [Google Scholar]
  5. Castagna M., Takai Y., Kaibuchi K., Sano K., Kikkawa U., Nishizuka Y. Direct activation of calcium-activated, phospholipid-dependent protein kinase by tumor-promoting phorbol esters. J Biol Chem. 1982 Jul 10;257(13):7847–7851. [PubMed] [Google Scholar]
  6. Creutz C. E., Dowling L. G., Sando J. J., Villar-Palasi C., Whipple J. H., Zaks W. J. Characterization of the chromobindins. Soluble proteins that bind to the chromaffin granule membrane in the presence of Ca2+. J Biol Chem. 1983 Dec 10;258(23):14664–14674. [PubMed] [Google Scholar]
  7. Fisher S. K., Holz R. W., Agranoff B. W. Muscarinic receptors in chromaffin cell cultures mediate enhanced phospholipid labeling but not catecholamine secretion. J Neurochem. 1981 Aug;37(2):491–497. doi: 10.1111/j.1471-4159.1981.tb00482.x. [DOI] [PubMed] [Google Scholar]
  8. Greenberg A., Zinder O. Alpha- and beta-receptor control of catecholamine secretion from isolated adrenal medulla cells. Cell Tissue Res. 1982;226(3):655–665. doi: 10.1007/BF00214792. [DOI] [PubMed] [Google Scholar]
  9. Gunther G. R. Effect of 12-O-tetradecanoyl-phorbol-13-acetate on Ca2+ efflux and protein discharge in pancreatic acini. J Biol Chem. 1981 Dec 10;256(23):12040–12045. [PubMed] [Google Scholar]
  10. Hutton J. C., Peshavaria M., Brocklehurst K. W. Phorbol ester stimulation of insulin release and secretory-granule protein phosphorylation in a transplantable rat insulinoma. Biochem J. 1984 Dec 1;224(2):483–490. doi: 10.1042/bj2240483. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kaibuchi K., Takai Y., Nishizuka Y. Cooperative roles of various membrane phospholipids in the activation of calcium-activated, phospholipid-dependent protein kinase. J Biol Chem. 1981 Jul 25;256(14):7146–7149. [PubMed] [Google Scholar]
  12. Katakami Y., Kaibuchi K., Sawamura M., Takai Y., Nishizuka Y. Synergistic action of protein kinase C and calcium for histamine release from rat peritoneal mast cells. Biochem Biophys Res Commun. 1984 Jun 15;121(2):573–578. doi: 10.1016/0006-291x(84)90220-1. [DOI] [PubMed] [Google Scholar]
  13. Knight D. E., Baker P. F. Stimulus-secretion coupling in isolated bovine adrenal medullary cells. Q J Exp Physiol. 1983 Jan;68(1):123–143. doi: 10.1113/expphysiol.1983.sp002691. [DOI] [PubMed] [Google Scholar]
  14. Knight D. E., Baker P. F. The phorbol ester TPA increases the affinity of exocytosis for calcium in 'leaky' adrenal medullary cells. FEBS Lett. 1983 Aug 22;160(1-2):98–100. doi: 10.1016/0014-5793(83)80944-2. [DOI] [PubMed] [Google Scholar]
  15. Kojima I., Lippes H., Kojima K., Rasmussen H. Aldosterone secretion: effect of phorbol ester and A23187. Biochem Biophys Res Commun. 1983 Oct 31;116(2):555–562. doi: 10.1016/0006-291x(83)90559-4. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Lord J. M., Ashcroft S. J. Identification and characterization of Ca2+-phospholipid-dependent protein kinase in rat islets and hamster beta-cells. Biochem J. 1984 Apr 15;219(2):547–551. doi: 10.1042/bj2190547. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Malaisse W. J., Sener A., Herchuelz A., Carpinelli A. R., Poloczek P., Winand J., Castagna M. Insulinotropic effect of the tumor promoter 12-O-tetradecanoylphorbol-13-acetate in rat pancreatic islets. Cancer Res. 1980 Oct;40(10):3827–3831. [PubMed] [Google Scholar]
  19. Minakuchi R., Takai Y., Yu B., Nishizuka Y. Widespread occurrence of calcium-activated, phospholipid-dependent protein kinase in mammalian tissues. J Biochem. 1981 May;89(5):1651–1654. doi: 10.1093/oxfordjournals.jbchem.a133362. [DOI] [PubMed] [Google Scholar]
  20. Niedel J. E., Kuhn L. J., Vandenbark G. R. Phorbol diester receptor copurifies with protein kinase C. Proc Natl Acad Sci U S A. 1983 Jan;80(1):36–40. doi: 10.1073/pnas.80.1.36. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Nishizuka Y. The role of protein kinase C in cell surface signal transduction and tumour promotion. Nature. 1984 Apr 19;308(5961):693–698. doi: 10.1038/308693a0. [DOI] [PubMed] [Google Scholar]
  22. Rink T. J., Sanchez A., Hallam T. J. Diacylglycerol and phorbol ester stimulate secretion without raising cytoplasmic free calcium in human platelets. Nature. 1983 Sep 22;305(5932):317–319. doi: 10.1038/305317a0. [DOI] [PubMed] [Google Scholar]
  23. Schatzman R. C., Raynor R. L., Fritz R. B., Kuo J. F. Purification to homogeneity, characterization and monoclonal antibodies of phospholipid-sensitive Ca2+-dependent protein kinase from spleen. Biochem J. 1983 Feb 1;209(2):435–443. doi: 10.1042/bj2090435. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Smith M. A., Vale W. W. Superfusion of rat anterior pituitary cells attached to Cytodex beads: validation of a technique. Endocrinology. 1980 Nov;107(5):1425–1431. doi: 10.1210/endo-107-5-1425. [DOI] [PubMed] [Google Scholar]
  25. Storer A. C., Cornish-Bowden A. Concentration of MgATP2- and other ions in solution. Calculation of the true concentrations of species present in mixtures of associating ions. Biochem J. 1976 Oct 1;159(1):1–5. doi: 10.1042/bj1590001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Takai Y., Kishimoto A., Iwasa Y., Kawahara Y., Mori T., Nishizuka Y. Calcium-dependent activation of a multifunctional protein kinase by membrane phospholipids. J Biol Chem. 1979 May 25;254(10):3692–3695. [PubMed] [Google Scholar]
  27. Takai Y., Kishimoto A., Kikkawa U., Mori T., Nishizuka Y. Unsaturated diacylglycerol as a possible messenger for the activation of calcium-activated, phospholipid-dependent protein kinase system. Biochem Biophys Res Commun. 1979 Dec 28;91(4):1218–1224. doi: 10.1016/0006-291x(79)91197-5. [DOI] [PubMed] [Google Scholar]
  28. Tanigawa K., Kuzuya H., Imura H., Taniguchi H., Baba S., Takai Y., Nishizuka Y. Calcium-activated, phospholipid-dependent protein kinase in rat pancreas islets of langerhans. Its possible role in glucose-induced insulin release. FEBS Lett. 1982 Feb 22;138(2):183–186. doi: 10.1016/0014-5793(82)80436-5. [DOI] [PubMed] [Google Scholar]
  29. Thams P., Capito K., Hedeskov C. J. Endogenous substrate proteins for Ca2+-calmodulin-dependent, Ca2+-phospholipid-dependent and cyclic AMP-dependent protein kinases in mouse pancreatic islets. Biochem J. 1984 Jul 1;221(1):247–253. doi: 10.1042/bj2210247. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Turgeon J. L., Ashcroft S. J., Waring D. W., Milewski M. A., Walsh D. A. Characteristics of the adenohypophyseal Ca2+-phospholipid-dependent protein kinase. Mol Cell Endocrinol. 1984 Feb;34(2):107–112. doi: 10.1016/0303-7207(84)90061-3. [DOI] [PubMed] [Google Scholar]
  31. Virji M. A., Steffes M. W., Estensen R. D. Phorbol myristate acetate: effect of a tumor promoter on insulin release from isolated rat islets of Langerhans. Endocrinology. 1978 Mar;102(3):706–711. doi: 10.1210/endo-102-3-706. [DOI] [PubMed] [Google Scholar]
  32. Wooten M. W., Wrenn R. W. Phorbol ester induces intracellular translocation of phospholipid/Ca2+-dependent protein kinase and stimulates amylase secretion in isolated pancreatic acini. FEBS Lett. 1984 Jun 11;171(2):183–186. doi: 10.1016/0014-5793(84)80484-6. [DOI] [PubMed] [Google Scholar]
  33. Wrenn R. W. Phospholipid-sensitive calcium-dependent protein kinase and its endogenous substrate proteins in rat pancreatic acinar cells. Life Sci. 1983 May 16;32(20):2385–2392. doi: 10.1016/0024-3205(83)90770-1. [DOI] [PubMed] [Google Scholar]
  34. Zawalich W., Brown C., Rasmussen H. Insulin secretion: combined effects of phorbol ester and A23187. Biochem Biophys Res Commun. 1983 Dec 16;117(2):448–455. doi: 10.1016/0006-291x(83)91221-4. [DOI] [PubMed] [Google Scholar]
  35. de Pont J. J., Fleuren-Jakobs A. M. Synergistic effect of A23187 and a phorbol ester on amylase secretion from rabbit pancreatic acini. FEBS Lett. 1984 May 7;170(1):64–68. doi: 10.1016/0014-5793(84)81369-1. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES