Potentiation by thrombin of the secretion of serotonin from permeabilized platelets equilibrated with Ca2+ buffers. Relationship to protein phosphorylation and diacylglycerol formation (original) (raw)

Abstract

After human platelets have been rendered permeable to small molecules by high voltage electric discharges, addition of buffered micromolar concentrations of Ca2+ causes an ATP-dependent secretion of dense granule serotonin [Knight & Scrutton (1980) Thromb. Res. 20, 437-446]. In the present study, platelets permeabilized by this technique were found to show an up to 10-fold increase in their sensitivity to Ca2+ after exposure to thrombin. In permeabilized platelets, as in the intact cells, release of serotonin was associated with the Ca2+-dependent phosphorylation of 47 000 and 20 000 Da polypeptides (P47 and P20). Thrombin markedly increased the phosphorylation of P47 in the presence of 0.1-1.0 microM-Ca2+ free but had a much smaller effect on phosphorylation of P20. Thrombin also stimulated the formation of 1,2-diacylglycerol in the presence of 0.1 microM-Ca2+ free and was even more effective with 1.0 microM-Ca2+ free, suggesting that receptor-activated hydrolysis of phosphoinositides to 1,2-diacylglycerol was preserved in permeabilized platelets and was potentiated by low intracellular concentrations of Ca2+. The increase in phosphorylation of P47 on addition of thrombin may therefore be accounted for by the stimulatory action of 1,2-diacylglycerol on Ca2+-activated, phospholipid-dependent protein kinase. However, in both the presence and absence of thrombin, higher Ca2+ concentrations were required for optimal secretion than for maximal phosphorylation of both P47 and P20, indicating that additional actions of Ca2+ and thrombin, perhaps also mediated by 1,2-diacylglycerol formation, may be involved in the release of serotonin.

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  1. ASTER R. H., JANDL J. H. PLATELET SEQUESTRATION IN MAN. I. METHODS. J Clin Invest. 1964 May;43:843–855. doi: 10.1172/JCI104970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Agranoff B. W., Murthy P., Seguin E. B. Thrombin-induced phosphodiesteratic cleavage of phosphatidylinositol bisphosphate in human platelets. J Biol Chem. 1983 Feb 25;258(4):2076–2078. [PubMed] [Google Scholar]
  3. Allan D., Michell R. H. Accumulation of 1,2-diacylglycerol in the plasma membrane may lead to echinocyte transformation of erythrocytes. Nature. 1975 Nov 27;258(5533):348–349. doi: 10.1038/258348a0. [DOI] [PubMed] [Google Scholar]
  4. BLIGH E. G., DYER W. J. A rapid method of total lipid extraction and purification. Can J Biochem Physiol. 1959 Aug;37(8):911–917. doi: 10.1139/o59-099. [DOI] [PubMed] [Google Scholar]
  5. Bell R. L., Majerus P. W. Thrombin-induced hydrolysis of phosphatidylinositol in human platelets. J Biol Chem. 1980 Mar 10;255(5):1790–1792. [PubMed] [Google Scholar]
  6. Berridge M. J. Phosphatidylinositol hydrolysis: a multifunctional transducing mechanism. Mol Cell Endocrinol. 1981 Nov;24(2):115–140. doi: 10.1016/0303-7207(81)90055-1. [DOI] [PubMed] [Google Scholar]
  7. Billah M. M., Lapetina E. G. Degradation of phosphatidylinositol-4,5-bisphosphate is insensitive to CA2+ mobilization in stimulated platelets. Biochem Biophys Res Commun. 1982 Nov 16;109(1):217–222. doi: 10.1016/0006-291x(82)91587-x. [DOI] [PubMed] [Google Scholar]
  8. Billah M. M., Lapetina E. G. Evidence for multiple metabolic pools of phosphatidylinositol in stimulated platelets. J Biol Chem. 1982 Oct 25;257(20):11856–11859. [PubMed] [Google Scholar]
  9. Daniel J. L., Molish I. R., Holmsen H. Myosin phosphorylation in intact platelets. J Biol Chem. 1981 Jul 25;256(14):7510–7514. [PubMed] [Google Scholar]
  10. Fabiato A., Fabiato F. Calculator programs for computing the composition of the solutions containing multiple metals and ligands used for experiments in skinned muscle cells. J Physiol (Paris) 1979;75(5):463–505. [PubMed] [Google Scholar]
  11. Feinman R. D., Detwiler T. C. Platelet secretion induced by divalent cation ionophores. Nature. 1974 May 10;249(453):172–173. doi: 10.1038/249172a0. [DOI] [PubMed] [Google Scholar]
  12. Haslam R. J., Lynham J. A., Fox J. E. Effects of collagen, ionophore A23187 and prostaglandin E1 on the phosphorylation of specific proteins in blood platelets. Biochem J. 1979 Feb 15;178(2):397–406. doi: 10.1042/bj1780397. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Haslam R. J., Lynham J. A. Relationship between phosphorylation of blood platelet proteins and secretion of platelet granule constituents. I. Effects of different aggregating agents. Biochem Biophys Res Commun. 1977 Jul 25;77(2):714–722. doi: 10.1016/s0006-291x(77)80037-5. [DOI] [PubMed] [Google Scholar]
  14. Imaoka T., Lynham J. A., Haslam R. J. Purification and characterization of the 47,000-dalton protein phosphorylated during degranulation of human platelets. J Biol Chem. 1983 Sep 25;258(18):11404–11414. [PubMed] [Google Scholar]
  15. Kaibuchi K., Takai Y., Sawamura M., Hoshijima M., Fujikura T., Nishizuka Y. Synergistic functions of protein phosphorylation and calcium mobilization in platelet activation. J Biol Chem. 1983 Jun 10;258(11):6701–6704. [PubMed] [Google Scholar]
  16. Kawahara Y., Takai Y., Minakuchi R., Sano K., Nishizuka Y. Phospholipid turnover as a possible transmembrane signal for protein phosphorylation during human platelet activation by thrombin. Biochem Biophys Res Commun. 1980 Nov 17;97(1):309–317. doi: 10.1016/s0006-291x(80)80169-0. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. 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]
  19. Knight D. E., Hallam T. J., Scrutton M. C. Agonist selectivity and second messenger concentration in Ca2+-mediated secretion. Nature. 1982 Mar 18;296(5854):256–257. doi: 10.1038/296256a0. [DOI] [PubMed] [Google Scholar]
  20. Knight D. E., Scrutton M. C. Cyclic nucleotides control a system which regulates Ca2+ sensitivity of platelet secretion. Nature. 1984 May 3;309(5963):66–68. doi: 10.1038/309066a0. [DOI] [PubMed] [Google Scholar]
  21. Knight D. E., Scrutton M. C. Direct evidence for a role for Ca2+ in amine storage granule secretion by human platelets. Thromb Res. 1980 Nov 15;20(4):437–446. doi: 10.1016/0049-3848(80)90282-0. [DOI] [PubMed] [Google Scholar]
  22. Lapetina E. G. Metabolism of inositides and the activation of platelets. Life Sci. 1983 May 2;32(18):2069–2082. doi: 10.1016/0024-3205(83)90094-2. [DOI] [PubMed] [Google Scholar]
  23. Lyons R. M., Stanford N., Majerus P. W. Thrombin-induced protein phosphorylation in human platelets. J Clin Invest. 1975 Oct;56(4):924–936. doi: 10.1172/JCI108172. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. MOLNAR J., LORAND L. Studies on apyrases. Arch Biochem Biophys. 1961 May;93:353–363. doi: 10.1016/0003-9861(61)90278-8. [DOI] [PubMed] [Google Scholar]
  25. Michell R. H. Inositol phospholipids and cell surface receptor function. Biochim Biophys Acta. 1975 Mar 25;415(1):81–47. doi: 10.1016/0304-4157(75)90017-9. [DOI] [PubMed] [Google Scholar]
  26. Michell R. H., Kirk C. J., Jones L. M., Downes C. P., Creba J. A. The stimulation of inositol lipid metabolism that accompanies calcium mobilization in stimulated cells: defined characteristics and unanswered questions. Philos Trans R Soc Lond B Biol Sci. 1981 Dec 18;296(1080):123–138. doi: 10.1098/rstb.1981.0177. [DOI] [PubMed] [Google Scholar]
  27. Naka M., Nishikawa M., Adelstein R. S., Hidaka H. Phorbol ester-induced activation of human platelets is associated with protein kinase C phosphorylation of myosin light chains. Nature. 1983 Dec 1;306(5942):490–492. doi: 10.1038/306490a0. [DOI] [PubMed] [Google Scholar]
  28. Persechini A., Hartshorne D. J. Cooperative behavior of smooth muscle myosin. Fed Proc. 1982 Oct;41(12):2868–2872. [PubMed] [Google Scholar]
  29. 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]
  30. Rink T. J., Smith S. W., Tsien R. Y. Cytoplasmic free Ca2+ in human platelets: Ca2+ thresholds and Ca-independent activation for shape-change and secretion. FEBS Lett. 1982 Nov 1;148(1):21–26. doi: 10.1016/0014-5793(82)81234-9. [DOI] [PubMed] [Google Scholar]
  31. Rittenhouse-Simmons S. Differential activation of platelet phospholipases by thrombin and ionophore A23187. J Biol Chem. 1981 May 10;256(9):4153–4155. [PubMed] [Google Scholar]
  32. Rittenhouse-Simmons S. Production of diglyceride from phosphatidylinositol in activated human platelets. J Clin Invest. 1979 Apr;63(4):580–587. doi: 10.1172/JCI109339. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Sano K., Takai Y., Yamanishi J., Nishizuka Y. A role of calcium-activated phospholipid-dependent protein kinase in human platelet activation. Comparison of thrombin and collagen actions. J Biol Chem. 1983 Feb 10;258(3):2010–2013. [PubMed] [Google Scholar]
  34. Takai Y., Minakuchi R., Kikkawa U., Sano K., Kaibuchi K., Yu B., Matsubara T., Nishizuka Y. Membrane phospholipid turnover, receptor function and protein phosphorylation. Prog Brain Res. 1982;56:287–301. doi: 10.1016/S0079-6123(08)63780-2. [DOI] [PubMed] [Google Scholar]
  35. Wallace W. C., Bensusan H. B. Protein phosphorylation in platelets stimulated by immobilized thrombin at 37 degrees and 4 degrees C. J Biol Chem. 1980 Mar 10;255(5):1932–1937. [PubMed] [Google Scholar]
  36. Wallace W. C., Bensusan H. B. The effects of platelet secretion inhibitors on protein phosphorylation. Biochim Biophys Acta. 1982 Oct 11;721(2):164–171. doi: 10.1016/0167-4889(82)90064-7. [DOI] [PubMed] [Google Scholar]
  37. White J. G., Rao G. H., Gerrard J. M. Effects of the lonophore A23187 on blood platelets I. Influence on aggregation and secretion. Am J Pathol. 1974 Nov;77(2):135–149. [PMC free article] [PubMed] [Google Scholar]
  38. 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]