Cardiac Sarcolemmal Vesicles: ATP-Dependent Ca Ion Transport and Inhibition of Protein Kinase Reactions by Amrinone

  • Roland Vetter
  • Hannelore Haase
  • Liane Will-Shahab
Part of the Developments in Cardiovascular Medicine book series (DICM, volume 102)


In the cardiac sarcolemma (SL), several Ca2+-transporting systems control cytosolic Ca2+ concentration which is linked to the contraction/relaxation cycle of the heart. 1 Ca2+ ions enter the myocardial cell during each excitation event via dihydropyridine-sensitive, voltage-operated Ca2+ channels.2,3 Both an electrogenic Na+/Ca2+ exchange process 4,5 and an ATP-driven Ca2+ pump participate in the extrusion of Ca2+ from the cell. These Ca2+ transport processes may be modulated by several membrane-associated protein kinases through protein phosphorylation reactions which are controlled by intracellular messengers like cyclic AMP, Ca2+, cyclic GMP and diacylglycerol.7–9 This modulation is a basic mechanism of mediation of the cellular response to a number of inotropic and chronotropic neurotransmitters, hormones and other agents. For example, it is well documented that the positive inotropic response to β-adrenergic agents includes SR Ca2+ pump activation via cyclic AMP and Ca2+/calmodulin-dependent phosphorylation of the membrane proteolipid phospholamban. 10,11 SL membrane contains several protein kinases which are controlled either by cyclic AMP, Ca2+/calmodulin or Ca2+/phospholipid. They catalyse the phosphorylation of different membrane substrates. 7,9 In SL, β-adrenergic stimulation or injection of cyclic AMP-dependent protein kinase into single cardiomyocytes is known to result in enhanced Ca2+ flux through slow Ca2+ channels. 12,13 Na+exchange in cardiac SL vesicles can be regulated by Ca2+/calmodulin-dependent phosphorylation/dephosphorylation reaction. 14


Dependent Protein Kinase Cardiac Sarcoplasmic Reticulum Cardiac Sarcolemma Heart Sarcolemma Cardiac Sarcolemmal Vesicle 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Dhalla, NS, Pierce GN, Pangia V, Singal PK, Beamish RE. Calcium movements in relation to heart function. Basic Res Cardiol 1982;77:117–139.PubMedCrossRefGoogle Scholar
  2. 2.
    Reuter H. Calcium channel modulation by neurotransmitters, enzymes and drugs. Nature 1984;301:567–574.Google Scholar
  3. 3.
    Tsien RW. Calcium channels in excitable cell membranes. Ann Rev Physiol 1983;45:341–358.CrossRefGoogle Scholar
  4. 4.
    Reuter H. Na-Ca countertransport in cardiac muscle. In: Martinosi A. (ed) Membranes and Transport. Vol 1. New York: Plenum Press, 1982. pp 623–631.Google Scholar
  5. 5.
    Sheu SS, Blaustein MP. Sodium/calcium exchange and regulation of cell calcium and contractility in cardiac muscle, with a note about vascular smooth muscle. In: Fozzard HA, Haber E, Jennings RB, Katz AM, Morgan HE. (eds) The Heart and Cardiovascular System. New York: Raven Press, 1986: pp 509–535.Google Scholar
  6. 6.
    Caroni P, Carafoli E. The Ca2+-pumping ATPase of the heart sarcolemma. Characterization, calmodulin dependence, partial purification. J Biol Chem 1981;256: 3263–3270.PubMedGoogle Scholar
  7. 7.
    Lamers JMJ. Cardiac sarcolemma calcium transport systems and their modulation by the second messengers cyclic AMP, calcium, and phosphoinositide products. In: Kidwai AM (ed) Sarcolemmal Biochemistry. Vol 2. Boca Raton, FL: CRC, 1987. pp 67–98.Google Scholar
  8. 8.
    Raeymaekers L, Hofmann F, Casteels R. Cyclic GMP-dependent protein kinase phosphoryiates phospholamban in isolated sarcoplasmic reticulum from cardiac and smooth muscle. Biochem J 1988;252:269–273.PubMedGoogle Scholar
  9. 9.
    Robinson-Steiner AM, Corbin JD. Protein phosphorylation in the heart. In: Fozzard HA, Haber E, Jennings RB, Katz AM, Morgan HE. (eds) The Heart and Cardiovascular System. New York: Raven Press, 1986: pp 887–910.Google Scholar
  10. 10.
    Tada M, Inui M. Regulation of the calcium transport by the ATPase-phospholamban system. J Mol Cell Cardiol 1983;15:565–575.PubMedCrossRefGoogle Scholar
  11. 11.
    Karczewski P, Vetter R, Holtzhauer M, Krause E-G. Indirect technique for the estimation of cAMP-dependent and Ca2+/calmodulin-dependent phospholamban phosphorylation state in canine heart in vivo. Biomed Biochem Acta 1986;45:227–231.Google Scholar
  12. 12.
    Sperelakis N. Cyclic AMP and phosphorylation in regulation of Ca2+ influx into myocardial cells and blockade by calcium antagonistic drugs. Am Heart J 1984;107:347–357.PubMedCrossRefGoogle Scholar
  13. 13.
    Ostrieder W, Brum G, Hescheler J, Trautwein W, Flockerzi V, Hofmann F. Injection of subunit of cAMP-dependent protein kinase into cardiac myocytes modulates Ca2+ current. Nature 1982;298:576–578.CrossRefGoogle Scholar
  14. 14.
    Caroni P, Carafdi E. The regulation of the Na+/Ca2+ exchanger of heart sarcolemma. Eur J Biochem 1983;132:451–460.PubMedCrossRefGoogle Scholar
  15. 15.
    Caroni P, Carafoli E. Regulation of Ca2+-pumping ATPase of heart sarcolemma by a phosphorylation — dephosphorylation process. J Biol Chem 1981;256:9371–9373.PubMedGoogle Scholar
  16. 16.
    Lamers JMJ, Stinis JT, Dejonge HR. On the role of cyclic AMP and Ca2+-calmodulin-dependent phosphorylation in the control of (Ca2+ +Mg2+)-ATPase of cardiac sarcolemma. FEBS Lett 1981; 127:139–143.PubMedCrossRefGoogle Scholar
  17. 17.
    Vetter R, Haase H, Will H. Potentiating effect of calmodulin and catalytic subunit of cyclic AMP-dependent protein kinase on ATP-dependent Ca2+-transport by cardiac sarcolemma. FEBS Lett 1982;148:326–330.PubMedCrossRefGoogle Scholar
  18. 18.
    Burgess WH, Jemiolo DK, Kretsinger RH. Interaction of calcium and calmodulin in the presence of sodium dodecyl sulfate. Biochim Biophys Acta 1980;623:257–270.PubMedGoogle Scholar
  19. 19.
    Peters KA, Demaille JG, Fischer EH. denosine-3,5,-monophosphate dependent protein kinase from bovine heart. Biochemistry 1977;16:5691–5697.PubMedCrossRefGoogle Scholar
  20. 20.
    Demaille JG, Peters KA, Fischer EH. Isolation and properties of the rabbit skeletal muscle protein inhibitor of adenosine 3,5,-monophosphate dependent protein kinases. Biochemistry 1977;16:3080–3086.PubMedCrossRefGoogle Scholar
  21. 21.
    Uchida T, Filburn CR. Affinity chromotography of protein kinase C — phorbol ester receptor on Polyacrylamide immobilized phosphatidylserine. J Biol Chem 1984;259:12311–12314.PubMedGoogle Scholar
  22. 22.
    Haase H, Wallukat G, Vetter R, Will H. Characterization of calcium antagonist receptors in highly purified porcine cardiac sarcolemma. Biomed Biochim Acta 1987;46:363–369.Google Scholar
  23. 23.
    Lowry OH, Rosebrough NJ, Farr AL, Randall J. Protein measurement with the folin reagent. J Biol Chem 1951; 193:265–275.PubMedGoogle Scholar
  24. 24.
    Vetter R, Will H. Sarcolemmal Na-Ca exchange and sarcoplasmic reticulum calcium uptake in developing chick heart. J Mol Cell Cardiol 1986;18:1267–275.PubMedCrossRefGoogle Scholar
  25. 25.
    Vetter R, Kemsies C, Schulze W. Sarcolemmal Na-Ca exchange and sarcoplasmic reticulum Ca-uptake in several cardiac preparations. Biomed Biochim Acta 1987;46:375–381.Google Scholar
  26. 26.
    Will-Shahab L, Krause E-G, Bartel S, Schulze W, Kuttner I. Reversible inhibition of adenylate cyclase activity in the ischemic myocardium. J Cardiovasc Pharmacol 1985;Suppl.5:S23–S27.Google Scholar
  27. 27.
    Wetzker R, Klinger R, Haase H, Vetter R, Böhmer FD. Fast activation of Ca2+-ATPase in plasma membranes from cardiac muscle and from ascites carcinoma cells: A possible function of endogenous calmodulin. Biomed Biochim Acta 1987;46:403–406.Google Scholar
  28. 28.
    Presti CF, Scott BT, Jones LR. Identification of an endogenous protein kinase C activity and its intrinsic 15-kilodalton substrate in purified canine cardiac sarcolemmal vesicles. J Biol Chem 1985;260:13879–13889.PubMedGoogle Scholar
  29. 29.
    Swank RT, Munkres KD. Molecular weight analysis of oligopeptides by electrophoresis in Polyacrylamide gel with sodium dodecyl sulfate. Anal Biochem 1971;39:462–477.PubMedCrossRefGoogle Scholar
  30. 30.
    Bartel S, Krause E-G, Wollenberger A. Assay of cyclic AMP-dependent protein kinase activity in canine myocardium: Effect of coronary artery ligation on the cytosolic enzyme. Biomed Biochlm Acta 1985;44:1303–1313.Google Scholar
  31. 31.
    Will H, Levchenko T, Kemsies C. Subunit analysis and cross-linking of phospholamban in cardiac sarcoplasmic reticulum and sarcolemma. In: Will-Shahab L, Krause E-G, Schulze W. Cellular and Molecular Aspects of the Regulation of the Heart. Berlin: Akademie-Verlag, 1982. pp 121–130.Google Scholar
  32. 32.
    Tuana BS, Dzurba A, Panagia V, Dhalla NS. Stimulation of heart sarcolemma calcium pump by calmodulin. Biochem Biophys Res Commun 1984;100:1245–1250.CrossRefGoogle Scholar
  33. 33.
    Iwasa Y, Hosey MM. Phosphorylation of cardiac sarcolemmal proteins by the calcium-activated phospholipid-dependent protein kinase. J Biol Chem 1984;259:534–540.PubMedGoogle Scholar
  34. 34.
    Yuan S, Sen AK. Characterization of the membrane-bound protein kinase C and its substrate proteins in canine cardiac sarcolemma. Biochim Biophys Acta 1986;886:152–161.PubMedCrossRefGoogle Scholar
  35. 35.
    Will H, Kuttner I, Vetter R, Will-Shahab L, Kemsies C. Early presence of phospholamban in developing chick heart. FEBS Lett 1983;155:326–330.PubMedCrossRefGoogle Scholar
  36. 36.
    Holtzhauer M, Sydow H, Will H. Characterization of endogenous phosphorylation in isolated cardiac sarcolemma. Gen Physiol Biophys 1983;2:437–446.PubMedGoogle Scholar
  37. 37.
    Jones L, Besch HR Jr, Fleming JW, McCounnaughey MM, Watanabe AM. Separation of vesicles of cardiac sarcolemmal from vesicles of sarcoplasmic reticulum. Comparative biochemical analysis of components. J Biol Chem 1979;254:530–539.PubMedGoogle Scholar
  38. 38.
    Limas CC. Phosphorylation of cardiac sarcoplasmic reticulum by a calcium-activated, phospholipid-dependent protein kinase. Biochem Biophys Res Commun 1980;96:1378–1383.PubMedCrossRefGoogle Scholar
  39. 39.
    Movsesian MA, Nishikawa M, Adelstein RS. Phosphorylation of phospholamban by calcium-activated, phospholipid-dependent protein kinase. Stimulation of cardiac sarcoplasmic reticulum calcium uptake. J Biol Chem 1984;259:8029–8031.PubMedGoogle Scholar
  40. 40.
    Alousi AA, Canter JM, Montenaro MJ, Fort PJ, Ferrari RA. Cardiotonic activity of milrinone, a new potent cardiac bipyridine, on isolated tissue from several animal species. J Cardiovasc Pharmacol 1983;5:804–872.PubMedCrossRefGoogle Scholar
  41. 41.
    Earl CQ, Linden J, Weglicki WB. Biochemical mechanisms for the inotropic efffects of the cardiotonic drug milrinone. J Cardiovasc Pharmacol 1988;8:864–872.Google Scholar
  42. 42.
    Earl CQ, Linden J, Weglicki WB. Inhibition of cyclic AMP-dependent protein kinase activity by the cardiotonic drugs amrinone and milrinone. Life Sci 1986;39:1901–1908.PubMedCrossRefGoogle Scholar
  43. 43.
    Hidaka H, Inagahi M, Kawamoto S, Sasaki Y. Isoquinolinesulfonamides, novel and potent inhibitors of cyclic nucleotide dependent protein kinase and protein kinase C. Biochemistry 1984;23:5036–5041.PubMedCrossRefGoogle Scholar
  44. 44.
    Edelson J, Stroshane R, Benzinger DP, Cody R, Benotti J, Hood JB Jr, Chatterjee K, Luczkowec C, Krebs C, Schwartz R. Pharmacokinetics of the bipyridines amrinone and milrinone. Circulation 1986;73:III/145–III/152.Google Scholar

Copyright information

© Kluwer Academic Publishers 1989

Authors and Affiliations

  • Roland Vetter
    • 1
  • Hannelore Haase
    • 1
  • Liane Will-Shahab
    • 1
  1. 1.Central Institute for Cardiovascular Research, Division of Cellular and Molecular CardiologyAcademy of Sciences of the GDRBerlin-BuchGermany

Personalised recommendations