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Ser16-, but not Thr17-phosphorylation of phospholamban influences frequency-dependent force generation in human myocardium

  • Cardiovascular System
  • Published:
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Abstract

β-Adrenoceptor/cAMP-dependent Ser16-phosphoryation as well as Ca2+-dependent Thr17-phosphorylation of phospholamban (PLB) influences SERCA 2a activity and thus myocardial contractility. To determine the cross-signaling between Ca2+ and cAMP pathways, the phosphorylation of Ser16-PLB and Thr17-PLB was studied at increasing stimulation frequencies as well as in the presence of β-adrenergic stimulation in isolated ventricular trabeculae from failing (dilative cardiomyopathy, DCM, heart transplants, n=9) and non-failing human myocardium (donor hearts, NF, n=9). In addition, we measured the intracellular Ca2+-transient (fura-2) at increasing stimulation frequencies (0.5–3.0 Hz). Protein expression of SERCA 2a and phospholamban was similar in DCM and NF. In DCM, diastolic [Ca2+]i was increased and systolic [Ca2+]i as well as Ser16 PLB-phosphorylation were decreased as compared to NF at 0.5 Hz. The positive force–frequency relationship in human non-failing myocardium was accompanied by a frequency-dependent increase in Ser16-PLB, but not Thr17-PLB phosphorylation. In DCM, Ser16-PLB as well as Thr17-PLB phosphorylation were not altered at higher stimulation frequencies. After application of isoprenaline (1 µM), a profound increase in Ser16-PLB phosphorylation was accompanied by a small increase in Thr17-PLB phosphorylation, only in NF. The frequency-dependent phosphorylation of Ser16-PLB may favor an increase in Ca2+ transient and force generation in humans. Cross talk signaling of Ser16/Thr17-PLB phosphorylation after β-adrenergic stimulation exists in non-failing, but not in failing human myocardium. The Ca2+-dependent CaM-kinase activity may be altered in human heart failure.

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Acknowledgements

We are indebted to all colleges of the Department of the Cardiothoracic Surgery of the University of Cologne (Director: Professor Dr. R. E. de Vivie) for providing us with human myocardial samples. The authors thank Mrs. Katja Rössler, Esra Köroglu and Kerstin Schenk for their excellent technical help. This study was supported by the Deutsche Forschungsgemeinschaft and the Köln Fortune Program/faculty of Medicine, University of Cologne. This work contains parts of the doctoral thesis of A.W.

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Authors and Affiliations

  1. Laboratory of Muscle Research and Molecular Cardiology, Department of Internal Medicine III, University of Cologne, Joseph-Stelzmann-Str. 9, 50924 , Cologne, Germany

    Klara Brixius, Annette Wollmer, Birgit Bölck & Robert H. G. Schwinger

  2. Clinic of Cardiothoracic Surgery, University of Cologne, Joseph-Stelzmann-Str. 9, 50924 , Cologne, Germany

    Uwe Mehlhorn

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  1. Klara Brixius
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  2. Annette Wollmer
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  5. Robert H. G. Schwinger
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Correspondence to Robert H. G. Schwinger.

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Brixius, K., Wollmer, A., Bölck, B. et al. Ser16-, but not Thr17-phosphorylation of phospholamban influences frequency-dependent force generation in human myocardium. Pflugers Arch - Eur J Physiol 447, 150–157 (2003). https://doi.org/10.1007/s00424-003-1163-3

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