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Expression of calcium channel subunits in the normal and diseased human myocardium

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Abstract

We investigated the expression of α1 and β subunits of the L-type Ca2+ channel on the protein level in cardiac preparations from normal human heart ventricles and from the hypertrophied septum of patients with hypertrophic obstructive cardiomyopathy (HOCM). 1,4-Dihydropyridine (DHP) binding and immunorecognition by polyclonal antibodies directed against the C-terminal amino acid sequences of the β2 and β3 subunits were used for detection and quantification of α1, β2, and β3 subunits. Bmax of high-affinity DHP binding was 35±2 fmol/mg protein in HOCM and 20±2 fmol/mg protein in normal human hearts (P<0.05). In rabbit hearts the anti-β2 subunit antibody immunoprecipitated 80% of the total amount of DHP-labeled Ca2+ channels present in the assay. Under identical experimental conditions 25% of labeled Ca2+ channels were recovered in the immunoprecipitates of both normal and HOCM ventricles. A similar partial immunoprecipitation was observed in pig hearts. Immunoblot analysis demonstrated that the β2 subunit was associated with the DHP receptor/Ca2+ channel in cardiac muscle of rabbit, pig, and human heart. In neither of these purified cardiac Ca2+ channels was the β3 subunit isoform detected. Our results suggest that both α1 and β2 subunit expression is upregulated in HOCM in a coordinate manner.

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Abbreviations

B max :

Maximal number of binding sites

DHP :

1,4-Dihydropyridine

HOCM :

Hypertrophic obstructive cardiomyopathy

NH :

Normal human heart

References

  1. Hullin R, Biel M, Flockerzi V, Hofmann F (1993) Tissue-specific expression of calcium channels. Trends Cardiovasc Med 3:48–54

    Google Scholar 

  2. Catterall WA, Striessnig J (1992) Receptor sites for Ca2+channel antagonists. Trends Pharamacol Sci 13:256–262

    Google Scholar 

  3. Wagner JA, Reynolds IJ, Weisman HF, Dudek P, Weisfeldt ML, Snyder SH (1986) Ca2+-antagonist receptors in cardiomyopathic hamster: selective increases in heart, muscle, brain. Science 232:515–517

    Google Scholar 

  4. Finkel MS, Patterson RE, Roberts WC, Smith TD, Keiser HR (1988) Calcium channel binding characteristics in the human heart. Am J Cardiol 62:1281–1284

    Google Scholar 

  5. Wagner JA, Sax FL, Weisman HF, Porterfield JK, McIntosh C, Weisfeldt ML, Snyder SH, Epstein SE (1989) Ca2+-antagonist receptors in the atrial tissue of patients with hypertrophic cardiomyopathy. N Engl J Med 320:755–761

    Google Scholar 

  6. Takahashi T, Allen PD, Lacro RV, Marks AR, Dennis AR, Schoen FJ, Grossman W, Marsh JD, Izumo S (1992) Expression of dihydropyridine receptor (Ca2+ channel) and calsequestrin genes in the myocardium of patients with end-stage heart failure. J Clin Invest 90:927–935

    Google Scholar 

  7. Rasmussen RP, Minobe W, Bristow MR (1990) Calcium antagonist binding sites in failing and nonfailing human ventricular myocardium. Biochem Pharmacol 39:691–696

    Google Scholar 

  8. Mewes T, Ravens U (1994) L-type calcium currents of human myocytes from ventricle of non-failing and failing human hearts and from atrium. J Mol Cell Cardiol 26:1307–1320

    Google Scholar 

  9. Sen L, Xu DC, Marks AR (1994) Developmental changes of Ca transport gene expression in genetically determined cardiomyopathic hamster hearts. Circulation 90, part IV, no 0186

  10. Neely A, Wei X, Olcese R, Birnbaumer L, Stefani E (1993) Potentiation by the β subunit of the ratio of the ionic current to the charge movement in the cardiac calcium channel. Science 262:575–578

    Google Scholar 

  11. Castellano A, Perez-Reyes E (1994) Molecular diversity of Ca2+ channel β subunits. Biochem Soc Trans 22:483–488

    Google Scholar 

  12. Perez-Reyes E, Castellano A, Kim HS, Bertrand P, Baggstrom E, Lacerda AE, Wei X, Birnbaumer L (1992) Cloning and expression of a cardiac/brain β subunit of the L-type calcium channel. J Biol Chem 267:1792–1797

    Google Scholar 

  13. Hullin R, Singer-Lahat D, Freichel M, Biel M, Dascal N, Hofmann F, Flockerzi V (1992) Calcium channel β subunit heterogeneity: functional expression of cloned cDNA from heart, aorta and brain. EMBO J 11:885–890

    Google Scholar 

  14. Collin T, Wang JJ, Nargeot J, Schwartz A (1993) Molecular cloning of three isoforms of the L-type voltage-dependent calcium channel β subunit from normal human heart. Circulation Res 72:1337–1344

    Google Scholar 

  15. Biel M, Hullin R, Freundner S, Singer D, Dascal N, Flockerzi V, Hofmann F (1991) Tissue-specific expression of high-voltage-activated dihydropyridine-sensitive L-type calcium channels. Eur J Biochem 200:81–88

    Google Scholar 

  16. Pragnell M, DeWaard M, Mori Y, Tanabe T, Snutch TP, Campbell KP (1994) Calcium channel β subunit binds to a conserved motif in the I–II cytoplasmic linker of the α1, subunit. Nature 368:67–70

    Google Scholar 

  17. DeWard M, Pragnell M, Campbell KP (1994) Ca2+ channel regulation by a conserved β subunit domain. Neuron 13:495–503

    Google Scholar 

  18. Witcher DR, De Waard M, Liu H, Pragnell M, Campbell KP (1995)Association of native Ca2+channel subunits with the α1 subunit interaction domain. J Biol Chem 270:18088–18093

    Google Scholar 

  19. Sakamoto J, Campbell KP (1991) A monoclonal antibody to the β subunit of the skeletal muscle dihydropyridine receptor immunoprecipitates the brain ω-conotoxin GVIA receptor. J Biol Chem 266:18914–18919

    Google Scholar 

  20. Haase H, Karczewski P, Beckert R, Krause EG (1993) Phosphorylation of the L-type calcium channel β subunit is involved in β-adrenergic signal transduction in canine myocardium. FEBS Lett 335:217–222

    Google Scholar 

  21. Haase H, Wallukat G, Flockerzi V, Nastainczyk W, Hofmann F (1994) Detection of skeletal muscle calcium channel subunits in cultured neonatal rat cardiac myocytes. Receptors Channels 2:41–52

    Google Scholar 

  22. Castellano A, Wei X, Birnbaumer L, Perez-Reyes E (1993) Cloning and expression of a third Ca2+ channel β subunit. J Biol Chem 268:3450–3455

    Google Scholar 

  23. Calovini T, Haase H, Morano I (1995) Steroid-hormone regulation of myosin subunit expression in smooth and cardiac muscle. J Cell Biochem 59:69–81

    Google Scholar 

  24. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193:265–275

    CAS  PubMed  Google Scholar 

  25. Haase H, Striessnig J, Holtzhauer M, Vetter R, Glossmann H (1991) A rapid procedure for the purification of cardiac 1,4-dihydropyridine receptors from porcine heart. Eur J Pharmacol 207:51–59

    Google Scholar 

  26. Karczewski P, Bartel S, Krause EG (1990) Differential sensitivity to isoprenaline of troponin I and phospholamban phosphorylation in isolated rat hearts. Biochem J 266:115–122

    Google Scholar 

  27. Striessnig J, Glossmann H (1991) Purification of L-type calcium channels drug receptors. Meth Neurosci 4:210–248

    Google Scholar 

  28. Schneider T, Regulla S, Nastainczyk W, Hofmann F (1992) Purification and structure of L-type calcium channels. In: Longstaff A, Revest P (eds) Methods in molecular biology protocols in molecular neurobiology, vol 13. Humana, Totowa, pp 273–286

    Google Scholar 

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Authors and Affiliations

  1. Max-Delbrück Center for Molecular Medicine, Robert-Rössle Strasse 10, D-13125, Berlin, Germany

    H. Hasse, A. Kresse, A. Hohaus & I. Morano

  2. Department of Thoracic Cardiovascular Surgery, Heinrich-Heine University, Düsseldorf, Germany

    H. D. Schulte

  3. German Heart Center, Berlin, Germany

    M. Maier & P. E. Lange

  4. Franz-Volhard Klinik, Humboldt University, Berlin, Germany

    K. J. Osterziel

Authors
  1. H. Hasse
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  2. A. Kresse
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  3. A. Hohaus
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  4. H. D. Schulte
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  5. M. Maier
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  6. K. J. Osterziel
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  7. P. E. Lange
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  8. I. Morano
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Hasse, H., Kresse, A., Hohaus, A. et al. Expression of calcium channel subunits in the normal and diseased human myocardium. J Mol Med 74, 99–104 (1996). https://doi.org/10.1007/BF00196785

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  • DOI: https://doi.org/10.1007/BF00196785

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