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. 1988 Jan 15;249(2):487–493. doi: 10.1042/bj2490487

Comparison of the effects of phorbol 12-myristate 13-acetate and prostaglandin E1 on calcium regulation in human platelets.

K Yoshida 1, F Stark 1, V T Nachmias 1
PMCID: PMC1148729  PMID: 2829859

Abstract

We compared the effects of phorbol 12-myristate 13-acetate (PMA) with those of prostaglandin E1 (PGE1) on the calcium transient in intact platelets and on 45Ca2+ uptake in saponin-treated platelets and microsomal fractions to determine the roles of protein kinase C and cyclic AMP in calcium sequestration. In intact platelets, PMA, like PGE1, stimulated the return of the calcium transient to resting values after a thrombin stimulus, but only the PGE1 effect was reversed by adrenaline. Both PMA and PGE1, when added before saponin, stimulated ATP-dependent 45Ca2+ uptake into the permeabilized platelets. Thrombin also stimulated 45Ca2+ uptake into saponin-treated platelets. Uptake of 45Ca2+ was increased in microsomal preparations from platelets pretreated with PMA or PGE1. PMA did not increase the cyclic AMP content of control or thrombin-treated platelets, and it induced a pattern of protein phosphorylation in 32P-labelled platelets different from that with PGE1. In correlation with the increased uptake of calcium in the saponin-treated preparation, we measured a rapid translocation of protein kinase C from supernatant to cell fraction after the addition of PMA. Our results suggest that activation of protein kinase C enhances calcium sequestration independently of an effect on cyclic AMP content in platelets. This activation could play a physiological role in the regulation of the calcium transient.

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Selected References

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

  1. Adunyah S. E., Dean W. L. Effect of phorbol esters and diacylglycerols on calcium transport by human platelet membranes. Cell Calcium. 1986 Jun;7(3):193–202. doi: 10.1016/0143-4160(86)90023-0. [DOI] [PubMed] [Google Scholar]
  2. Authi K. S., Evenden B. J., Crawford N. Metabolic and functional consequences of introducing inositol 1,4,5-trisphosphate into saponin-permeabilized human platelets. Biochem J. 1986 Feb 1;233(3):707–718. doi: 10.1042/bj2330707. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brass L. F., Joseph S. K. A role for inositol triphosphate in intracellular Ca2+ mobilization and granule secretion in platelets. J Biol Chem. 1985 Dec 5;260(28):15172–15179. [PubMed] [Google Scholar]
  4. Brass L. F., Laposata M., Banga H. S., Rittenhouse S. E. Regulation of the phosphoinositide hydrolysis pathway in thrombin-stimulated platelets by a pertussis toxin-sensitive guanine nucleotide-binding protein. Evaluation of its contribution to platelet activation and comparisons with the adenylate cyclase inhibitory protein, Gi. J Biol Chem. 1986 Dec 25;261(36):16838–16847. [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. Chantler P. D., Sellers J. R., Szent-Györgyi A. G. Cooperativity in scallop myosin. Biochemistry. 1981 Jan 6;20(1):210–216. doi: 10.1021/bi00504a035. [DOI] [PubMed] [Google Scholar]
  7. Connolly T. M., Lawing W. J., Jr, Majerus P. W. Protein kinase C phosphorylates human platelet inositol trisphosphate 5'-phosphomonoesterase, increasing the phosphatase activity. Cell. 1986 Sep 12;46(6):951–958. doi: 10.1016/0092-8674(86)90077-2. [DOI] [PubMed] [Google Scholar]
  8. Fox J. E., Say A. K., Haslam R. J. Subcellular distribution of the different platelet proteins phosphorylated on exposure of intact platelets to ionophore A23187 or to prostaglandin E1. Possible role of a membrane phosphopolypeptide in the regulation of calcium-ion transport. Biochem J. 1979 Dec 15;184(3):651–661. doi: 10.1042/bj1840651. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Grynkiewicz G., Poenie M., Tsien R. Y. A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem. 1985 Mar 25;260(6):3440–3450. [PubMed] [Google Scholar]
  10. Inagaki M., Kawamoto S., Hidaka H. Serotonin secretion from human platelets may be modified by Ca2+-activated, phospholipid-dependent myosin phosphorylation. J Biol Chem. 1984 Dec 10;259(23):14321–14323. [PubMed] [Google Scholar]
  11. Iwasa Y., Hosey M. M. Phosphorylation of cardiac sarcolemma proteins by the calcium-activated phospholipid-dependent protein kinase. J Biol Chem. 1984 Jan 10;259(1):534–540. [PubMed] [Google Scholar]
  12. Katada T., Gilman A. G., Watanabe Y., Bauer S., Jakobs K. H. Protein kinase C phosphorylates the inhibitory guanine-nucleotide-binding regulatory component and apparently suppresses its function in hormonal inhibition of adenylate cyclase. Eur J Biochem. 1985 Sep 2;151(2):431–437. doi: 10.1111/j.1432-1033.1985.tb09120.x. [DOI] [PubMed] [Google Scholar]
  13. Kranias E. G. Regulation of Ca2+ transport by cyclic 3',5'-AMP-dependent and calcium-calmodulin-dependent phosphorylation of cardiac sarcoplasmic reticulum. Biochim Biophys Acta. 1985 Feb 21;844(2):193–199. doi: 10.1016/0167-4889(85)90090-4. [DOI] [PubMed] [Google Scholar]
  14. Käser-Glanzmann R., Gerber E., Lüscher E. F. Regulation of the intracellular calcium level in human blood platelets: cyclic adenosine 3',5'-monophosphate dependent phosphorylation of a 22,000 dalton component in isolated Ca2+-accumulating vesicles. Biochim Biophys Acta. 1979 Dec 12;558(3):344–347. doi: 10.1016/0005-2736(79)90271-2. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. Le Peuch C. J., Le Peuch D. A., Katz S., Demaille J. G., Hincke M. T., Bredoux R., Enouf J., Levy-Toledano S., Caen J. Regulation of calcium accumulation and efflux from platelet vesicles. Possible role for cyclic-AMP-dependent phosphorylation and calmodulin. Biochim Biophys Acta. 1983 Jun 23;731(3):456–464. doi: 10.1016/0005-2736(83)90041-x. [DOI] [PubMed] [Google Scholar]
  17. Menashi S., Authi K. S., Carey F., Crawford N. Characterization of the calcium-sequestering process associated with human platelet intracellular membranes isolated by free-flow electrophoresis. Biochem J. 1984 Sep 1;222(2):413–417. doi: 10.1042/bj2220413. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Molina y Vedia L. M., Lapetina E. G. Phorbol 12,13-dibutyrate and 1-oleyl-2-acetyldiacylglycerol stimulate inositol trisphosphate dephosphorylation in human platelets. J Biol Chem. 1986 Aug 15;261(23):10493–10495. [PubMed] [Google Scholar]
  19. Movsesian M. A., Nishikawa M., Adelstein R. S. Phosphorylation of phospholamban by calcium-activated, phospholipid-dependent protein kinase. Stimulation of cardiac sarcoplasmic reticulum calcium uptake. J Biol Chem. 1984 Jul 10;259(13):8029–8032. [PubMed] [Google Scholar]
  20. 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]
  21. Nishizuka Y. Studies and perspectives of protein kinase C. Science. 1986 Jul 18;233(4761):305–312. doi: 10.1126/science.3014651. [DOI] [PubMed] [Google Scholar]
  22. Pollock W. K., Rink T. J., Irvine R. F. Liberation of [3H]arachidonic acid and changes in cytosolic free calcium in fura-2-loaded human platelets stimulated by ionomycin and collagen. Biochem J. 1986 May 1;235(3):869–877. doi: 10.1042/bj2350869. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Pollock W. K., Sage S. O., Rink T. J. Stimulation of Ca2+ efflux from fura-2-loaded platelets activated by thrombin or phorbol myristate acetate. FEBS Lett. 1987 Jan 5;210(2):132–136. doi: 10.1016/0014-5793(87)81322-4. [DOI] [PubMed] [Google Scholar]
  24. Rittenhouse S. E., Sasson J. P. Mass changes in myoinositol trisphosphate in human platelets stimulated by thrombin. Inhibitory effects of phorbol ester. J Biol Chem. 1985 Jul 25;260(15):8657–8660. [PubMed] [Google Scholar]
  25. Tada M., Ohmori F., Yamada M., Abe H. Mechanism of the stimulation of Ca2+-dependent ATPase of cardiac sarcoplasmic reticulum by adenosine 3':5'-monophosphate-dependent protein kinase. Role of the 22,000-dalton protein. J Biol Chem. 1979 Jan 25;254(2):319–326. [PubMed] [Google Scholar]
  26. Tapley P. M., Murray A. W. Evidence that treatment of platelets with phorbol ester causes proteolytic activation of Ca2+-activated, phospholipid-dependent protein kinase. Eur J Biochem. 1985 Sep 2;151(2):419–423. doi: 10.1111/j.1432-1033.1985.tb09118.x. [DOI] [PubMed] [Google Scholar]
  27. Uratsuji Y., Nakanishi H., Takeyama Y., Kishimoto A., Nishizuka Y. Activation of cellular protein kinase C and mode of inhibitory action of phospholipid-interacting compounds. Biochem Biophys Res Commun. 1985 Jul 31;130(2):654–661. doi: 10.1016/0006-291x(85)90467-x. [DOI] [PubMed] [Google Scholar]
  28. Watson S. P., Lapetina E. G. 1,2-Diacylglycerol and phorbol ester inhibit agonist-induced formation of inositol phosphates in human platelets: possible implications for negative feedback regulation of inositol phospholipid hydrolysis. Proc Natl Acad Sci U S A. 1985 May;82(9):2623–2626. doi: 10.1073/pnas.82.9.2623. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Yoshida K., Dubyak G., Nachmias V. T. Rapid effects of phorbol ester on platelet shape change, cytoskeleton and calcium transient. FEBS Lett. 1986 Oct 6;206(2):273–278. doi: 10.1016/0014-5793(86)80995-4. [DOI] [PubMed] [Google Scholar]
  30. Zavoico G. B., Feinstein M. B. Cytoplasmic Ca2+ in platelets is controlled by cyclic AMP: antagonism between stimulators and inhibitors of adenylate cyclase. Biochem Biophys Res Commun. 1984 Apr 30;120(2):579–585. doi: 10.1016/0006-291x(84)91294-4. [DOI] [PubMed] [Google Scholar]

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