Regulation and functional significance of phospholipase D in myocardium

  • Yvonne E. G. Eskildsen-Helmond
  • Han A. A. Van Heugten
  • Jos M. J. Lamers
Part of the Developments in Molecular and Cellular Biochemistry book series (DMCB, volume 17)


There is now clear evidence that receptor-dependent phospholipase D is present in myocardium. This novel signal transduction pathway provides an alternative source of 1,2-diacylglycerol, which activates isoforms of protein kinase C. The members of the protein kinase C family respond differently to various combinations of Ca2+, phosphatidylserine, molecular species of 1,2-diacylglycerol and other membrane phospholipid metabolites including free fatty acids. Protein kinase C isozymes are responsible for phosphorylation of specific cardiac substrate proteins that may be involved in regulation of cardiac contractility, hypertrophic growth, gene expression, ischemic preconditioning and electrophysiological changes. The initial product of phospholipase D, phosphatidic acid, may also have a second messenger role. As in other tissues, the question how the activity of phospholipase D is controlled by agonists in myocardium is controversial. Agonists, such as endothelin-1, atrial natriuretic factor and angiotensin II that are shown to activate phospholipase D, also potently stimulate phospholipase C-β in myocardium. PMA stimulation of protein kinase C inactivates phospholipase C and strongly activates phospholipase D and this is probably a major mechanism by which agonists that promote phosphatidyl-4,5-bisphosphate hydrolysis secondary activate phosphatidylcholine-hydrolysis. On the other hand, one group has postulated that formation of phosphatidic acid secondary activates phosphatidyl-4,5-bisphosphate hydrolysis in cardiomyocytes. Whether GTP-binding proteins directly control phospholipase D is not clearly established in myocardium. Phospholipase D activation may also be mediated by an increase in cytosolic free Ca2+ or by tyrosine-phosphorylation.

Key words

phospholipase D signaltransduction myocardium cardiomyocytes protein kinase C phospholipase C phosphatidic acid phosphatidylethanol hypertrophy ischemic preconditioning inotrophy 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Brown JH, Martinson JH: Phosphoinositide-generated second messengers in cardiac signal transduction. Trends Cardiovasc Med 2: 209–213, 1992PubMedCrossRefGoogle Scholar
  2. 2.
    De Jonge HW, Van Heugten HAA, Lamers JMJ: Signal transduction by the phosphatidylinositol cycle in myocardium. J Mol Cell Cardiol 27: 93–106, 1995PubMedCrossRefGoogle Scholar
  3. 3.
    Van Heugten HAA, De Jonge HW, Bezstarosti K, Sharma HS, Verdouw PD, Lamers JMJ: Intracellular signalling and genetic reprogramming during agonist-induced hypertrophy of cardiomyocytes. Ann N Y Acad Sci 752: 343–352, 1995PubMedCrossRefGoogle Scholar
  4. 4.
    Chien KR, Knowlton KU, Zhu H, Chien S: Regulation of cardiac gene expression during myocardial growth and hypertrophy: molecular studies of an adaptive physiologic response. FASEB J 5: 3037–3046, 1991PubMedGoogle Scholar
  5. 5.
    Van Heugten HAA, De Jonge HW, Goedbloed MA, Bezstarosti K, Sharma HS, Verdouw PD, Lamers JMJ: Intracellular signalling and genetic reprogramming during development of hypertrophy in cultured cardiomyocytes. In: N.S. Dhalla, P.K. Singal, R.E. Beamish (eds). Heart Hypertrophy and Failure, Kluwer Academic Publishers, Boston 1995, pp 79–92CrossRefGoogle Scholar
  6. 6.
    Edwards DR: Cell signalling and the control of gene transcription. Trends Pharmacol Sci 15: 239–244, 1994PubMedCrossRefGoogle Scholar
  7. 7.
    Blumer KJ, Johnson GL: Diversity in function and regulation of MAP kinase pathways. Trends Pharmacol Sci 19: 236–240, 1994Google Scholar
  8. 8.
    Exton JH: Signalling through phosphatidylcholine breakdown. J Biol Chem 265: 1–4, 1990PubMedGoogle Scholar
  9. 9.
    Shukla SD, Halenda SP: Phospholipase D in cell signalling and relationship to phospholipase C. Life Sci 48: 851–866, 1991PubMedCrossRefGoogle Scholar
  10. 10.
    Kiss Z: Effects of phorbolester on phospholipid metabolism. Prog Lipid Res 29: 141–166, 1990PubMedCrossRefGoogle Scholar
  11. 11.
    Meij JTA, Lamers JMJ: Phorbolester inhibits α1-adrenoceptor mediated phosphoinositide breakdown in cardiomyocytes. J Mol Cell Cardiol 21: 661–668, 1989PubMedCrossRefGoogle Scholar
  12. 12.
    Meij JTA, Bezstarosti K, Panagia V, Lamers JMJ: Phorbolester and the actions of phosphatidyl 4,5-bisphosphate specific phospholipase C and protein kinase C in microsomes prepared from cultured cardiomyocytes. Mol Cell Biochem 105: 37–47, 1989Google Scholar
  13. 13.
    Van Heugten HAA, Bezstarosti K, Dekkers DHW, Lamers JMJ: Homologous desensitization of the endothelin-1 receptor mediated phosphoinositide response in cultured neonatal rat cardiomyocytes. J Mol Cell Cardiol 25: 41–52, 1993PubMedCrossRefGoogle Scholar
  14. 14.
    Berridge M, Irvine RF: Inositol phosphates and cell signalling. Nature 341: 197–205, 1989PubMedCrossRefGoogle Scholar
  15. 15.
    Kanfer JN: Phospholipase D and the base exchange enzyme. In: D.E. Vance (ed.). Phosphatidylcholine Metabolism. CRC Press, Florida, 1989, pp 65–86Google Scholar
  16. 16.
    Billah MM, Anthes JC: The regulation and cellular functions of phosphatidylcholine hydrolysis. Biochem J 269: 281–291, 1990PubMedGoogle Scholar
  17. 17.
    Billah MM: Phospholipase D and cell signalling. Current Opinion in Immunology 5: 114–123, 1993PubMedCrossRefGoogle Scholar
  18. 18.
    Thompson NT, Garland LG, Bonser RW: Phospholipase D: Regulation and functional significance. Adv Pharmacol 24: 199–238, 1993PubMedCrossRefGoogle Scholar
  19. 19.
    Christie WW: Lipid Analysis, 2nd edition. Pergamon Press, Oxford, 1982, pp 155–166Google Scholar
  20. 20.
    Exton JH: Phosphatidylcholine breakdown and signal transduction. Biochim Biophys Acta 1212: 26–42, 1994PubMedGoogle Scholar
  21. 21.
    Saito M, Bourque E, Kanfer JN: Phosphatidohydrolase and base-exchange activity of commercial phospholipase D. Arch Biochem Biophys 164: 420–428, 1974PubMedCrossRefGoogle Scholar
  22. 22.
    Lindmar R, Löffelholz K, Sandmann J: On the mechanism of muscarinic hydrolysis of choline phospholipids. Biochem Pharmacol 37: 4689–4695, 1988PubMedCrossRefGoogle Scholar
  23. 23.
    Panagia V, Ou C, Taira Y, Dai J, Dhalla NS: Phospholipase D activity in subcellular membranes of rat ventricular myocardium. Biochim Biophys Acta 1064 (2): 242–50, 1991PubMedCrossRefGoogle Scholar
  24. 24.
    Berridge MJ: Inositol trisphosphate and diacylglycerol: Two interacting second messengers. Ann Rev Biochem 56: 159–193, 1987PubMedCrossRefGoogle Scholar
  25. 25.
    Thompson NT, Bonser RW, Garland LG: Receptor-coupled phospholipase D and its inhibition. Trends Pharmacol Sci 12: 404–408, 1991PubMedCrossRefGoogle Scholar
  26. 26.
    Doglio A, Dani C, Grimaldi P, Ailhaud G: Growth hormone stimulates c-fos gene expression by means of protein kinase C without increasing inositol lipid turnover. Proc Natl Acad Sci USA 86: 1148–1152, 1989PubMedCrossRefGoogle Scholar
  27. 27.
    Pfeffer LM, Strulovici B, Saltiel AR: Interferon-a selectively activates the β isoform of protein kinase C through phosphatidylcholine hydrolysis. Proc Natl Acad Sci USA 87: 6537–6541, 1990PubMedCrossRefGoogle Scholar
  28. 28.
    De Jonge HW, Van Heugten HAA, Bezstarosti K, Lamers JMJ: Distinct α1-adrenergic agonist- and ET-1 evoked phosphoinositide cycle responses in cultured neonatal rat cardiomyocytes. Biochem Biophys Res Commun 203: 422–429, 1994PubMedCrossRefGoogle Scholar
  29. 29.
    Lamers JMJ, Eskildsen-Helmond YEG, Resink AM, de Jonge HW, Bezstarosti K, Sharma HS, van Heugten HAA: Endothelin-1 induced phospholipase C-β and D and protein kinase C-isoenzyme signalling leading to hypertrophy in rat cardiomyocytes. J Cardiovasc Pharm 26 (Suppl. 3): S100–S103, 1995Google Scholar
  30. 30.
    Hongping Y, Wolf RA, Kurz T, Corr PB: Phosphatidic acid increases in response to noradrenaline and endothelin-1 in adult rabbit ventricular myocytes. Cardiovasc Res 28: 18128–1834, 1994Google Scholar
  31. 31.
    Kurz T, Wolf RA, Corr PB: Phosphatidic acid stimulates inositol 1,4,5-triphosphate production in adult cardiac myocytes. Circ Res 72: 701–706, 1993PubMedGoogle Scholar
  32. 32.
    Jones AW, Shukla SD, Geisbuhler BB: Stimulation of phospholipase D activity and phosphatidic acid production by norepinephrine in rat aorta. Am J Physiol 264: C609–C616, 1993PubMedGoogle Scholar
  33. 33.
    Sadoshima J, Izumo S: Signal transduction pathways of angiotensin II-induced cfos gene expression in cardiac myocytes in vitro. Circ Res 73: 424–438, 1993PubMedGoogle Scholar
  34. 34.
    Baldini PM, Incerpi S, Zannetti A, De Vito P, Luly P: Selective activation by atrial natriuretic factor of phosphatidylcholine-specific phospholipase activities in purified heart muscle plasma membranes. J Mol Cell Cardiol 26: 1691–1700, 1994PubMedCrossRefGoogle Scholar
  35. 36.
    Booz GW, Taher MM, Baker KM, Singer HA: Angiotensin II induces phosphatidic acid formation in neonatal rat cardiac fibroblasts: Evaluation of the roles of phospholipase C and D. Mol Cell Biochem 141: 135–143, 1994PubMedCrossRefGoogle Scholar
  36. 35.
    Putney JW Jr, Weiss SJ, Van De Walle CM, Haddas RA: Is phosphatidic acid a calcium ionophore under neurohumoral control? Nature 284: 345–347, 1980PubMedCrossRefGoogle Scholar
  37. 37.
    Sharma HS, Van Heugten HA, Goedbloed MA, Verdouw PD, Lamers JMJ: Angiotensin II induced expression of transcription factors preceded increase in transforming growth factor-beta mRNA in neonatal cardiac fibroblasts. Biochim Biophys Res Commun 205: 105–112, 1994CrossRefGoogle Scholar
  38. 38.
    Tilly BC, Lambrechts AC, Tertoolen LG, de Laat SW, Moolenaar WH: Regulation of phosphoinositide hydrolysis induced by histamine and guanine nucleotides in human HeLa carcinoma cells. Calcium and pH dependence and inhibitory role of protein kinase C. FEBS-Lett 265: 80–84, 1990PubMedCrossRefGoogle Scholar
  39. 39.
    Van Heugten HAA, De Jonge HW, Bezstarosti K, Lamers JMJ: Calcium and the endothelin-1 and α1-adrenergic stimulated phosphatidylinositol cycle in cultured rat cardiomyocytes. J Mol Cell Cardiol 26: 1081–1093, 1994PubMedCrossRefGoogle Scholar
  40. 40.
    McDonough PM, Goldstein D, Brown JH: Elevation of cytoplasmic calcium concentration stimulates hydrolysis of phosphatidylinositol biphosphate in chick heart cells: Effect of sodium channel activators. Mol Pharmacol 33: 310–315, 1988PubMedGoogle Scholar
  41. 41.
    Jones LG, Brown JH: Guanine nucleotide-regulated inositol polyphosphate production in adult rat cardiomyocytes. In: W. A. Clark, R.S. Decker, T.K. Borg (eds). Biology of Isolated Adult Cardiac myocytes. Elsevier Publishing Co., New York, 1988, pp 257–260Google Scholar
  42. 42.
    Billah MM, Pai J-K, Mullmann TJ, Egan RW, Siegel MI: Regulation of phospholipase D in H-60 granulocytes. J Biol Chem 264: 9069–9076, 1989PubMedGoogle Scholar
  43. 43.
    Lui Y, Geisbuhler B, Jones AW: Activation of multiple mechanisms including phospholipase D by endothelin-1 in rat aorta. Am J Physiol 262: C941–C949, 1992Google Scholar
  44. 44.
    Pessin MS, Raben DM: Molecular species analysis of 1,2-diglycerides stimulated by α-thrombin in cultured fibroblasts. J Biol Chem 264: 8729–8738, 1989PubMedGoogle Scholar
  45. 45.
    Van Blitterswijk WJ, Hilkman H, De Widt J, Van der Bend RL: Phospholipid metabolism in bradykinin-stimulated human fibroblasts. J Biol Chem 266: 10344–10350, 1991PubMedGoogle Scholar
  46. 46.
    Divecha N, Irvine RF: Phospholipid signalling. Cell 80: 269–278, 1995PubMedCrossRefGoogle Scholar
  47. 47.
    Wakelam MJO, Pettitt TR, Kaur P, Briscoe CP, Stewart A, Paul A, Paterson A, Cross MJ, Gardner SD, Currie S, MacNulty EE, Plevin R, Cook SJ: Phosphatidylcholine hydrolysis: a multiple messenger generating system. In: B.L. Brown, R.M. Dobson (eds). Advances in Second Messenger and Phosphoprotein Research Vol 28. Raven Press, New York, 1993, pp 73–80Google Scholar
  48. 48.
    Wang P, Anthes JC, Siegel MI, Egan RW and Billah MM: Existence of cytosolic phospholipase D: Identification and comparison with membrane-bound enzyme. J Biol Chem 266: 14877–14880, 1991PubMedGoogle Scholar
  49. 49.
    Chalifour RJ, Kanfer JN: Microsomal phospholipase D of rat brain and lung tissues. Biochem Biophys Res Commun 96: 742–747, 1980PubMedCrossRefGoogle Scholar
  50. 50.
    Taki T, Kanfer JN: Phospholipase D from rat brain. Meth Enzymol 71: 746–750, 1981PubMedCrossRefGoogle Scholar
  51. 51.
    Cockcroft S: G-protein-regulated phospholipase C, D and A2-mediated signalling in neutrophils. Biochim Biophys Acta 1113: 135–160, 1992PubMedGoogle Scholar
  52. 52.
    Geny B, Cockcroft S: Synergistic activation of phospholipase D by protein kinase C- and G-protein-mediated pathways in streptolysin O-permeabilized HL60 cells. Biochem J 284: 531–538, 1992PubMedGoogle Scholar
  53. 53.
    De Jonge HW, Atsma DE, Van der Valk-Kokshoorn EJM, Van Heugten HAA, Van der Laarse A, Lamers JMJ: Alpha-adrenergic agonist and endothelin-1 induced intracellular Ca2+ response in the presence of a Ca2+ entry blocker in cultured rat ventricular myocytes. Cell Calcium 18: 515–525, 1995PubMedCrossRefGoogle Scholar
  54. 54.
    Lee CH, Park D, Wu D, Rhee SG, Simon MI: Members of the Gq α subunit gene family activate phospholipase C β isozymes. J Biol Chem 267: 16044–16047, 1992PubMedGoogle Scholar
  55. 55.
    Sadoshima J, Izumo S: Mechanical stretch rapidly activates multiple signal transduction pathways in cardiac myocytes: potential involvement of an autocrine/paracrine mechanism. EMBO J 12, no. 4: 1681–1692, 1993PubMedGoogle Scholar
  56. 56.
    Morgan HE, Baker KM: Cardiac hypertrophy. Mechanical, neural, and endocrine dependence. Circulation 83: 13–25, 1991PubMedGoogle Scholar
  57. 57.
    Simpson PC, Kariya K-I, Karns LR, Long CS, Karliner JS: Adrenergic hormones and control of cardiac myocyte growth. Mol Cell Biochem 104: 35–43, 1991PubMedCrossRefGoogle Scholar
  58. 58.
    Gu X, Bishop P: Increased protein kinase C and isozyme redistribution in pressure-overload cardiac hypertrophy in the rat. Circ Res 75: 926–931, 1994PubMedGoogle Scholar
  59. 59.
    Bogoyevitch MA, Parker PJ, Sugden PH: Characterization of protein kinase C isotype expression in adult rat heart. A protein kinase C-γ is a major isotype present, and it is activated by phorbol esters, epinephrine and endothelin. Circ Res 72: 757–767, 1993aGoogle Scholar
  60. 60.
    Mochly-Rosen D, Henrich CJ, Cheever L, Khaner H, Simpson PC: A protein kinase C isozyme is translocated to cytoskeletal elements on activation. Cell Regulation 1: 693–706, 1990PubMedGoogle Scholar
  61. 61.
    Steinberg SF, Goldberg M, Rybin VO: Protein kinase C isoform diversity in the heart. J Mol Cell Cardiol 27: 141–153, 1995PubMedCrossRefGoogle Scholar
  62. 62.
    Puceat M, Hilal-Dandan R, Strulovici B, Brunton LL, Brown JH: Differential regulation of protein kinase C isoforms in isolated neonatal and adult cardiomyocytes. J Biol Chem 269: 16938–16944, 1994PubMedGoogle Scholar
  63. 63.
    Nakamura S, Nishizuka Y: Lipid mediators and protein kinase C activation for the intracellular signalling network. J Biochem 115: 1029–1034, 1994PubMedGoogle Scholar
  64. 64.
    Cohen MV, Downey JM: Ischaemic preconditioning: can the protection be bottled? The Lancet 342: 6, 1993CrossRefGoogle Scholar
  65. 65.
    Moraru II, Popescu LM, Maulik N, Liu X, Das DK: Phospholipase D signalling in ischemic heart. Biochim Biophys Acta 1139: 148–154, 1992PubMedGoogle Scholar
  66. 66.
    Yasuda M, Kohno M, Tahara A, Stagone H, Tode I, Akioka K, Teragaki M, Oku H, Takenchi K, Takede T: Circulatory immunoreactive endothelin in ischemic heart disease. Am Heart J 119: 801–809, 1990PubMedCrossRefGoogle Scholar
  67. 67.
    Van Heugten HAA, Bezstarosti K, Lamers JMJ: Endothelin-1 and phenylephrine-induced activation of the phosphoinositide cycle increases cell injury of cultured cardiomyocytes exposed to hypoxia/reoxygenation. J Mol Cell Cardiol 26: 1513–1524, 1994PubMedCrossRefGoogle Scholar
  68. 68.
    Lawson CS, Downey JM: preconditioning: state of art myocardial protection. Cardiovasc Res: 542–550, 1993Google Scholar
  69. 69.
    Mitchell MB, Xionzhong M, Lihua A, Brown JM, Harken AH, Banerjee A: Preconditioning of isolated rat heart is mediated by protein kinase C. Circ Res 76: 73–81, 1995PubMedGoogle Scholar
  70. 70.
    Langer GA, Rich TL: Phospholipase D produces increased contractile force in rabbit ventricular muscle. Circ Res 56: 146–149, 1985PubMedGoogle Scholar
  71. 71.
    Burt JM, Rick TL, Langer GA: Phospholipase D increases cell surface Ca2+ binding and positive inotropy in rat heart. Am J Physiol 247: H880–H885, 1984PubMedGoogle Scholar
  72. 72.
    Philipson KD, Nishimoto AY: Stimulation of Na+-Ca2+ exchange in cardiac sarcolemmal vesicles by phospholipase D. J Biol Chem 259: 16–19, 1984PubMedGoogle Scholar

Copyright information

© Kluwer Academic Publishers 1996

Authors and Affiliations

  • Yvonne E. G. Eskildsen-Helmond
    • 1
  • Han A. A. Van Heugten
    • 1
  • Jos M. J. Lamers
    • 1
  1. 1.Department of Biochemistry, Cardiovascular Research Institute (COEUR), Faculty of Medicine and Health SciencesErasmus University RotterdamRotterdamThe Netherlands

Personalised recommendations