Abstract
The effects of phorbol 12-myristate 13-acetate (PMA) on catecholamine secretion and protein phosphorylation from intact and digitonin-treated chromaffin cells were investigated. PMA (10-300 nM), an activator of protein kinase C, caused a slow Ca2+-dependent release of catecholamine from intact chromaffin cells that was potentiated by the Ca2+ ionophore ionomycin. PMA also enhanced secretion induced by Ba2+. In cells with plasma membranes rendered permeable by digitonin to Ca2+, ATP, and protein, PMA (100 nM) enhanced Ca2+-dependent secretion approximately 70% at 0.5 microM Ca2+ and 30% at 10 microM Ca2+. PMA enhanced the maximal response to Ca2+ approximately 25% and decreased the Ca2+ concentration required for half-maximal secretion approximately 30%. The effects of PMA on chromaffin cells were associated with a 2- to 3-fold increase in the phosphorylation of a 56-kDa protein that may be tyrosine hydroxylase. Other proteins were phosphorylated to a lesser extent. The experiments suggest that PMA increases protein kinase activity and secretion in chromaffin cells and raise the possibility that protein kinase C modulates catecholamine secretion in chromaffin cells.
Full text
PDF![930](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8743/397161/5034928d5820/pnas00343-0304.png)
![931](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8743/397161/4dec9139a818/pnas00343-0305.png)
![932](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8743/397161/f12a9aa668e3/pnas00343-0306.png)
![933](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8743/397161/a4d580e956c3/pnas00343-0307.png)
![934](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8743/397161/2d12cb8a2d21/pnas00343-0308.png)
Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- 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]
- 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]
- Dunn L. A., Holz R. W. Catecholamine secretion from digitonin-treated adrenal medullary chromaffin cells. J Biol Chem. 1983 Apr 25;258(8):4989–4993. [PubMed] [Google Scholar]
- Haycock J. W., Meligeni J. A., Bennett W. F., Waymire J. C. Phosphorylation and activation of tyrosine hydroxylase mediate the acetylcholine-induced increase in catecholamine biosynthesis in adrenal chromaffin cells. J Biol Chem. 1982 Nov 10;257(21):12641–12648. [PubMed] [Google Scholar]
- Holz R. W., Senter R. A., Frye R. A. Relationship between Ca2+ uptake and catecholamine secretion in primary dissociated cultures of adrenal medulla. J Neurochem. 1982 Sep;39(3):635–646. doi: 10.1111/j.1471-4159.1982.tb07940.x. [DOI] [PubMed] [Google Scholar]
- Kikkawa U., Takai Y., Minakuchi R., Inohara S., Nishizuka Y. Calcium-activated, phospholipid-dependent protein kinase from rat brain. Subcellular distribution, purification, and properties. J Biol Chem. 1982 Nov 25;257(22):13341–13348. [PubMed] [Google Scholar]
- Kilpatrick D. L., Slepetis R. J., Corcoran J. J., Kirshner N. Calcium uptake and catecholamine secretion by cultured bovine adrenal medulla cells. J Neurochem. 1982 Feb;38(2):427–435. doi: 10.1111/j.1471-4159.1982.tb08647.x. [DOI] [PubMed] [Google Scholar]
- Kishimoto A., Takai Y., Mori T., Kikkawa U., Nishizuka Y. Activation of calcium and phospholipid-dependent protein kinase by diacylglycerol, its possible relation to phosphatidylinositol turnover. J Biol Chem. 1980 Mar 25;255(6):2273–2276. [PubMed] [Google Scholar]
- 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]
- Knight D. E., Kesteven N. T. Evoked transient intracellular free Ca2+ changes and secretion in isolated bovine adrenal medullary cells. Proc R Soc Lond B Biol Sci. 1983 May 23;218(1211):177–199. doi: 10.1098/rspb.1983.0033. [DOI] [PubMed] [Google Scholar]
- Kraft A. S., Anderson W. B. Phorbol esters increase the amount of Ca2+, phospholipid-dependent protein kinase associated with plasma membrane. Nature. 1983 Feb 17;301(5901):621–623. doi: 10.1038/301621a0. [DOI] [PubMed] [Google Scholar]
- O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
- Ueda T., Greengard P. Adenosine 3':5'-monophosphate-regulated phosphoprotein system of neuronal membranes. I. Solubilization, purification, and some properties of an endogenous phosphoprotein. J Biol Chem. 1977 Jul 25;252(14):5155–5163. [PubMed] [Google Scholar]
- Waymire J. C., Bennett W. F., Boehme R., Hankins L., Gilmer-Waymire K., Haycock J. W. Bovine adrenal chromaffin cells: high-yield purification and viability in suspension culture. J Neurosci Methods. 1983 Apr;7(4):329–351. doi: 10.1016/0165-0270(83)90026-2. [DOI] [PubMed] [Google Scholar]
- Wilson S. P., Kirshner N. Calcium-evoked secretion from digitonin-permeabilized adrenal medullary chromaffin cells. J Biol Chem. 1983 Apr 25;258(8):4994–5000. [PubMed] [Google Scholar]
- Wise B. C., Raynor R. L., Kuo J. F. Phospholipid-sensitive Ca2+-dependent protein kinase from heart. I. Purification and general properties. J Biol Chem. 1982 Jul 25;257(14):8481–8488. [PubMed] [Google Scholar]
- Yamanishi J., Takai Y., Kaibuchi K., Sano K., Castagna M., Nishizuka Y. Synergistic functions of phorbol ester and calcium in serotonin release from human platelets. Biochem Biophys Res Commun. 1983 Apr 29;112(2):778–786. doi: 10.1016/0006-291x(83)91529-2. [DOI] [PubMed] [Google Scholar]