Calcium uptake by sarcoplasmic reticulum and its modulation by cAMP-dependent phosphorylation in normal and failing human myocardium.

  • M. A. Movsesian
Conference paper


ATP-dependent, oxalate-supported Ca2+ uptake by cardiac sarcoplasmic reticulum was examined in microsomes prepared from left-ventricular free wall myocardium obtained from the explanted failing hearts of transplant recipients with idiopathic dilated cardiomyopathy and the non-failing hearts of kidney donors for whose hearts no suitable recipients were available. There were no significant differences between the two groups with respect to values for Vmax, K0.5 (for Ca2+) or nHill of basal Ca2+ uptake. The stimulation of Ca2+ uptake associated with cAMP-dependent phosphorylation of phospholamban could be reproduced by incubation of microsomes with a monoclonal antibody to phospholamban. Stimulation resulted from a decrease in K0.5, with no changes in Vmax or nHill. The magnitude of stimulation of Ca2+ uptake following incubation with anti-phospholamban monoclonal antibody was identical in preparations from normal and failing hearts. Finally, sarcoplasmic reticulum-associated cGMP-inhibited cAMP phosphodiesterase activity in these preparations was characterised. Measurement of steady-state kinetics and pharmacologic sensitivity indicated that this activity was functionally homogeneous. Preparations from failing and non- failing hearts did not differ with respect to either values for Vmax and Km or susceptibility to inhibition by the cilostamide derivative OPC 3911. These observations indicate that abnormalities in the regulation of intracellular [Ca2+] in failing human myocardium cannot be ascribed to changes in the level or function of the Ca2 + -transporting ATPase, phospholamban or cGMP-inhibited cAMP phosphodiesterase in the sarcoplasmic reticulum.

Key words

Heart failure sarcoplasmic reticulum Ca2+ uptake Ca2+ ATPase phospholamban phosphodiesterase phosphodiesterase inhibitors cAMP cGMP cilostamide rolipram 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Böhm M, Diet F, Feiler G, Kemkes B, Kreuzer E, Weinhold C, Erdmann E (1988) Subsensitivity of the failing human heart to isoprenaline and milrinone is related to β- adrenoceptor downregulation. J Cardiovasc Pharm 12: 726–732CrossRefGoogle Scholar
  2. 2.
    Bristow MR, Hershberger RE, Port JD, Minobe W, Rasmussen R (1989) β r and β 2- adrenergic receptor-mediated adenylate cyclase stimulation in nonfailing and failing human ventricular myocardium. Mol Pharm 35: 295–303Google Scholar
  3. 3.
    Feldman AM, Cates AE, Veazey WB, Hershberger RE, Bristow MR, Baughman KL, Baumgartner WA, Van Dop C (1988) Increase of the 40,000-mol wt pertussis toxin substrate in the failing human heart. J Clin Invest 82: 189–197PubMedCrossRefGoogle Scholar
  4. 4.
    Feldman MD, Copelas L, Gwathmey JK, Phillips P, Warren SE, Schoen FJ, Grossman W, Morgan JP (1987) Deficient production of cyclic AMP: pharmacologic evidence of an important cause of contractile dysfunction in patients with end-stage heart failure. Circulation 75: 331–339PubMedCrossRefGoogle Scholar
  5. 5.
    Gilbert EM, Hershberger RE, Mealey PC, Yolkman K, Weichman RJ, Menlove RL, Movsesian MA, Bristow MR. Pharmacologic and hemodynamic effects of combined β- agonist stimulation and phospodiesterase inhibition in the failing human heart (submitted for publication)Google Scholar
  6. 6.
    Gwathmey JK, Copelas L, MacKinnon R, Schoen FJ, Feldman MD, Grossman W, Morgan JP (1987) Abnormal intracellular calcium handling in myocardium from patients with end-stage heart failure. Circ Res 61: 70–76PubMedGoogle Scholar
  7. 7.
    Limas CJ, Olivari MT, Goldenberg IF, Levine TB, Benditt DG, Simon A (1987) Calcium uptake by cardiac sarcoplasmic reticulum in human dilated cardiomyopathy. Cardiovsac Res 21: 601–605CrossRefGoogle Scholar
  8. 8.
    Movsesian MA, Bristow MR, Krall J (1989) Calcium uptake by cardiac sarcoplasmic reticulum from patients with idiopathic dilated cardiomyopathy. Circ Res 65: 1141–1144PubMedGoogle Scholar
  9. 9.
    Movsesian MA, Colyer J, Wang JH, Krall J (1990) Phospholamban-mediated stimulation of Ca2 + uptake in cardiac sarcoplasmic reticulum from normal and failing hearts. J Clin Invest 85: 1698–1792PubMedCrossRefGoogle Scholar
  10. 10.
    Movsesian MA, Leveille C, Krall J, Colyer J, Wang JH, Campbell KP (1990) Identification and characterization of proteins in human cardiac sarcoplasmic reticulum. J Mol Cell Cardiol 22: 1477–1485PubMedCrossRefGoogle Scholar
  11. 11.
    Movsesian MA, Smith CJ, Krall J, Bristow M, Manganiello VC (1991) Sarcoplasmic reticulum-associated cAMP phosphodiesterase activity in normal and failing human hearts. J Clin Invest 88: 15–19PubMedCrossRefGoogle Scholar
  12. 12.
    Peeters GA, Hlady V, Bridge JHB, Barry WH (1987) Simultaneous measurement of calcium transients and motion in cultured ventricular cells. Am J Physiol 253: H1400–H1408PubMedGoogle Scholar
  13. 13.
    Suzuki T, Wang JH (1986) Stimulation of bovine cardiac sarcoplasmic reticulum Ca2 + pump and blocking of phospholamban phosphorylation and dephosphorylation by a phospholamban monoclonal antibody. J Biol Chem 261: 7018–7023PubMedGoogle Scholar
  14. 14.
    Tada M, Kadoma M, Inui M, Fujii J-I (1988) Regulation of Ca2+-pump from cardiac sarcoplasmic reticulum. Methods Enzymol 157: 107–154PubMedCrossRefGoogle Scholar
  15. 15.
    Weishaar RE, Kobylarz-Singer DC, Quade MM, Steffen RP, Kaplan HR (1987) Multiple molecular forms of phosphodiesterase and the regulation of cardiac muscle contractility. J Cyclic Nucleotide Protein Phosphorylation Res 11: 513–527Google Scholar
  16. 16.
    Weishaar RE, Kobylarz-Singer DC, Steffen RP, Kaplan HR (1987) Subclasses of cyclic AMP-specific phosphodiesterase in left ventricular muscle and their involvement in regulating myocardial contractility. Circ Res 61: 539–547PubMedGoogle Scholar

Copyright information

© Dr. Dietrich Steinkopff Verlag GmbH & Co.KG, Darmstadt 1992

Authors and Affiliations

  • M. A. Movsesian
    • 1
    • 2
  1. 1.Cardiology DivisionUniversity of Utah Medical CenterSalt Lake CityUSA
  2. 2.Cardiology DivisionUniversity of Utah Medical CenterSalt Lake CityUSA

Personalised recommendations