Regulation of Sodium-Calcium Exchanger Activity by Creatine Kinase

  • Ya-Chi Yang
  • Lung-Sen KaoEmail author
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 961)


It has been shown that in rat heart NCX1 exists in a macromolecular ­complex including PKA, PKA-anchoring protein, PKC, and phosphatases PP1 and PP2A. In addition, several lines of evidence suggest that the interactions of the exchanger with other molecules are closely associated with its function in regulation of [Ca2+]i. NCX contains a large intracellular loop (NCXIL) that is responsible for regulating NCX activity. We used the yeast two-hybrid method to screen a human heart cDNA library and found that the C-terminal region of sarcomeric mitochondrial creatine kinase (sMiCK) interacted with NCX1IL. Among the four creatine kinase (CK) isozymes, both sMiCK and the muscle-type cytosolic creatine kinase (CKM) co-immunoprecipitated with NCX1. Both sMiCK and CKM were able to produce a recovery in the decreased NCX1 activity that was lost under energy-compromised conditions. This regulation is mediated through a putative PKC phosphorylation site of sMiCK and CKM. The catalytic activity of sMiCK and CKM is not required for their regulation of NCX1 activity. Our results suggest a novel mechanism for the regulation of NCX1 activity and a novel role for CK.


Sodium-calcium exchanger1 (NCX1) Creatine kinase Energy-compromised conditions NCX1 macromolecular complex 



This research was originally published in the Journal of Biological Chemistry; Yang, Y.-C., Fann, M.-J., Chang, W.-H., Tai, L.-H., Jiang, J.-H., and Kao, L.-S. (2010) Regulation of sodium-calcium exchanger activity by creatine kinase under energy-compromised conditions. J. Biol. Chem. Vol. 285, 28275–28285 © the American Society for Biochemistry and Molecular Biology.


  1. G.M. Besserer, M. Ottolia, D.A. Nicoll, V. Chaptal, D. Cascio, K.D. Philipson, J. Abramson, The second Ca2+-binding domain of the Na+ Ca2+ exchanger is essential for regulation: crystal structures and mutational analysis. Proc. Natl. Acad. Sci. U. S. A. 104, 18467–18472 (2007)PubMedCrossRefGoogle Scholar
  2. M.P. Blaustein, W.J. Lederer, Sodium/calcium exchange: its physiological implications. Physiol. Rev. 79, 763–854 (1999)PubMedGoogle Scholar
  3. M.P. Blaustein, W.F. Goldman, G. Fontana, B.K. Krueger, E.M. Santiago, T.D. Steele, D.N. Weiss, P.J. Yarowsky, Physiological roles of the sodium-calcium exchanger in nerve and muscle. Ann. N. Y. Acad. Sci. 639, 254–274 (1991)PubMedCrossRefGoogle Scholar
  4. Y.J. Chern, S.H. Chueh, Y.J. Lin, C.M. Ho, L.S. Kao, Presence of Na+/Ca2+ exchange activity and its role in regulation of intracellular calcium concentration in bovine adrenal chromaffin cells. Cell Calcium 13, 99–106 (1992)PubMedCrossRefGoogle Scholar
  5. C.H. Cho, S.S. Kim, M.J. Jeong, C.O. Lee, H.S. Shin, The Na+-Ca2+ exchanger is essential for embryonic heart development in mice. Mol. Cells 10, 712–722 (2000)PubMedGoogle Scholar
  6. M. Condrescu, J.P. Gardner, G. Chernaya, J.F. Aceto, C. Kroupis, J.P. Reeves, ATP-dependent regulation of sodium-calcium exchange in Chinese hamster ovary cells transfected with the bovine cardiac sodium-calcium exchanger. J. Biol. Chem. 270, 9137–9146 (1995)PubMedCrossRefGoogle Scholar
  7. R. DiPolo, L. Beauge, Phosphoarginine stimulation of Na  +  -Ca2+ exchange in squid axons–a new pathway for metabolic regulation? J. Physiol. 487(Pt 1), 57–66 (1995)PubMedGoogle Scholar
  8. R. DiPolo, L. Beauge, Metabolic pathways in the regulation of invertebrate and vertebrate Na+/Ca2+ exchange. Biochim. Biophys. Acta 1422, 57–71 (1999)PubMedCrossRefGoogle Scholar
  9. R. DiPolo, L. Beauge, Sodium/calcium exchanger: influence of metabolic regulation on ion carrier interactions. Physiol. Rev. 86, 155–203 (2006)PubMedCrossRefGoogle Scholar
  10. R. DiPolo, G. Berberian, D. Delgado, H. Rojas, L. Beauge, A novel 13 kDa cytoplasmic soluble protein is required for the nucleotide (MgATP) modulation of the Na+/Ca2+ exchange in squid nerve fibers. FEBS Lett. 401, 6–10 (1997)PubMedCrossRefGoogle Scholar
  11. A.E. Doering, D.A. Nicoll, Y. Lu, L. Lu, J.N. Weiss, K.D. Philipson, Topology of a functionally important region of the cardiac Na+/Ca2+ exchanger. J. Biol. Chem. 273, 778–783 (1998)PubMedCrossRefGoogle Scholar
  12. M. Eder, K. Fritz-Wolf, W. Kabsch, T. Wallimann, U. Schlattner, Crystal structure of human ubiquitous mitochondrial creatine kinase. Proteins 39, 216–225 (2000a)PubMedCrossRefGoogle Scholar
  13. M. Eder, M. Stolz, T. Wallimann, U. Schlattner, A conserved negatively charged cluster in the active site of creatine kinase is critical for enzymatic activity. J. Biol. Chem. 275, 27094–27099 (2000b)PubMedGoogle Scholar
  14. B.N. Eigel, H. Gursahani, R.W. Hadley, Na+/Ca2+ exchanger plays a key role in inducing apoptosis after hypoxia in cultured guinea pig ventricular myocytes. Am. J. Physiol. Heart Circ. Physiol. 287, H1466–H1475 (2004)PubMedCrossRefGoogle Scholar
  15. R.A. Haworth, A.V. Biggs, Effect of ATP depletion on kinetics of Na+/Ca2+ exchange-mediated Ca2+ influx in Na+-loaded heart cells. J. Mol. Cell. Cardiol. 29, 503–514 (1997)PubMedCrossRefGoogle Scholar
  16. D.W. Hilgemann, Cytoplasmic ATP-dependent regulation of ion transporters and channels: mechanisms and messengers. Annu. Rev. Physiol. 59, 193–220 (1997)PubMedCrossRefGoogle Scholar
  17. A. Ikari, H. Sakai, N. Takeguchi, Protein kinase C-mediated up-regulation of Na+/Ca2+-exchanger in rat hepatocytes determined by a new Na+/Ca2+-exchanger inhibitor, KB-R7943. Eur. J. Pharmacol. 360, 91–98 (1998)PubMedCrossRefGoogle Scholar
  18. T. Iwamoto, S. Wakabayashi, M. Shigekawa, Growth factor-induced phosphorylation and activation of aortic smooth muscle Na+/Ca2+ exchanger. J. Biol. Chem. 270, 8996–9001 (1995)PubMedCrossRefGoogle Scholar
  19. T. Iwamoto, Y. Pan, S. Wakabayashi, T. Imagawa, H.I. Yamanaka, M. Shigekawa, Phosphorylation-dependent regulation of cardiac Na+/Ca2+ exchanger via protein kinase C. J. Biol. Chem. 271, 13609–13615 (1996)PubMedCrossRefGoogle Scholar
  20. T. Iwamoto, S. Kita, A. Uehara, Y. Inoue, Y. Taniguchi, I. Imanaga, M. Shigekawa, Structural domains influencing sensitivity to isothiourea derivative inhibitor KB-R7943 in cardiac Na+/Ca2+ exchanger. Mol. Pharmacol. 59, 524–531 (2001)PubMedGoogle Scholar
  21. L.S. Kao, Calcium homeostasis in digitonin-permeabilized bovine chromaffin cells. J. Neurochem. 51, 221–227 (1988)PubMedCrossRefGoogle Scholar
  22. L.S. Kao, N.S. Cheung, Mechanism of calcium transport across the plasma membrane of bovine chromaffin cells. J. Neurochem. 54, 1972–1979 (1990)PubMedCrossRefGoogle Scholar
  23. P. Kofuji, W.J. Lederer, D.H. Schulze, Mutually exclusive and cassette exons underlie alternatively spliced isoforms of the Na/Ca exchanger. J. Biol. Chem. 269, 5145–5149 (1994)PubMedGoogle Scholar
  24. I. Komuro, K.E. Wenninger, K.D. Philipson, S. Izumo, Molecular cloning and characterization of the human cardiac Na+/Ca2+ exchanger cDNA. Proc. Natl. Acad. Sci. U. S. A. 89, 4769–4773 (1992)PubMedCrossRefGoogle Scholar
  25. D.O. Levitsky, D.A. Nicoll, K.D. Philipson, Identification of the high affinity Ca2+-binding domain of the cardiac Na+-Ca2+ exchanger. J. Biol. Chem. 269, 22847–22852 (1994)PubMedGoogle Scholar
  26. Z. Li, D.A. Nicoll, A. Collins, D.W. Hilgemann, A.G. Filoteo, J.T. Penniston, J.N. Weiss, J.M. Tomich, K.D. Philipson, Identification of a peptide inhibitor of the cardiac sarcolemmal Na+-Ca2+ exchanger. J. Biol. Chem. 266, 1014–1020 (1991)PubMedGoogle Scholar
  27. Z. Li, S. Matsuoka, L.V. Hryshko, D.A. Nicoll, M.M. Bersohn, E.P. Burke, R.P. Lifton, K.D. Philipson, Cloning of the NCX2 isoform of the plasma membrane Na+-Ca2+ exchanger. J. Biol. Chem. 269, 17434–17439 (1994)PubMedGoogle Scholar
  28. G. Lin, Y. Liu, K.M. MacLeod, Regulation of muscle creatine kinase by phosphorylation in normal and diabetic hearts. Cell. Mol. Life Sci. 66, 135–144 (2009)PubMedCrossRefGoogle Scholar
  29. P.S. Liu, L.S. Kao, Na+-dependent Ca2+ influx in bovine adrenal chromaffin cells. Cell Calcium 11, 573–579 (1990)PubMedCrossRefGoogle Scholar
  30. S.O. Marx, J. Kurokawa, S. Reiken, H. Motoike, J. D’Armiento, A.R. Marks, R.S. Kass, Requirement of a macromolecular signaling complex for beta adrenergic receptor modulation of the KCNQ1-KCNE1 potassium channel. Science 295, 496–499 (2002)PubMedCrossRefGoogle Scholar
  31. S. Matsuoka, D.A. Nicoll, Z. He, K.D. Philipson, Regulation of cardiac Na+-Ca2+ exchanger by the endogenous XIP region. J. Gen. Physiol. 109, 273–286 (1997)PubMedCrossRefGoogle Scholar
  32. J.A. Mattiello, K.B. Margulies, V. Jeevanandam, S.R. Houser, Contribution of reverse-mode sodium-calcium exchange to contractions in failing human left ventricular myocytes. Cardiovasc. Res. 37, 424–431 (1998)PubMedCrossRefGoogle Scholar
  33. E.D. Moore, E.F. Etter, K.D. Philipson, W.A. Carrington, K.E. Fogarty, L.M. Lifshitz, F.S. Fay, Coupling of the Na+/Ca2+ exchanger, Na+/K+ pump and sarcoplasmic reticulum in smooth muscle. Nature 365, 657–660 (1993)PubMedCrossRefGoogle Scholar
  34. D.A. Nicoll, B.D. Quednau, Z. Qui, Y.R. Xia, A.J. Lusis, K.D. Philipson, Cloning of a third mammalian Na+-Ca2+ exchanger, NCX3. J. Biol. Chem. 271, 24914–24921 (1996)PubMedCrossRefGoogle Scholar
  35. M. Ottolia, D.A. Nicoll, K.D. Philipson, Roles of two Ca2+-binding domains in regulation of the cardiac Na+-Ca2+ exchanger. J. Biol. Chem. 284, 32735–32741 (2009)PubMedCrossRefGoogle Scholar
  36. C.Y. Pan, L.S. Kao, Catecholamine secretion from bovine adrenal chromaffin cells: the role of the Na+/Ca2+ exchanger and the intracellular Ca2+ pool. J. Neurochem. 69, 1085–1092 (1997)PubMedCrossRefGoogle Scholar
  37. C.Y. Pan, Y.S. Chu, L.S. Kao, Molecular study of the Na+/Ca2+ exchanger in bovine adrenal chromaffin cells. Biochem. J. 336(Pt 2), 305–310 (1998)PubMedGoogle Scholar
  38. C.Y. Pan, L.L. Tsai, J.H. Jiang, L.W. Chen, L.S. Kao, The co-presence of Na+/Ca2+-K+ exchanger and Na+/Ca2+ exchanger in bovine adrenal chromaffin cells. J. Neurochem. 107, 658–667 (2008)PubMedCrossRefGoogle Scholar
  39. M.V. Pulina, R. Rizzuto, M. Brini, E. Carafoli, Inhibitory interaction of the plasma membrane Na+/Ca2+ exchangers with the 14-3-3 proteins. J. Biol. Chem. 281, 19645–19654 (2006)PubMedCrossRefGoogle Scholar
  40. B.D. Quednau, D.A. Nicoll, K.D. Philipson, Tissue specificity and alternative splicing of the Na+/Ca2+ exchanger isoforms NCX1, NCX2, and NCX3 in rat. Am. J. Physiol. 272, C1250–C1261 (1997)PubMedGoogle Scholar
  41. B.D. Quednau, D.A. Nicoll, K.D. Philipson, The sodium/calcium exchanger family-SLC8. Pflugers Arch. 447, 543–548 (2004)PubMedCrossRefGoogle Scholar
  42. X. Ren, D.A. Nicoll, K.D. Philipson, Helix packing of the cardiac Na+-Ca2+ exchanger: proximity of transmembrane segments 1, 2, and 6. J. Biol. Chem. 281, 22808–22814 (2006)PubMedCrossRefGoogle Scholar
  43. M. Reppel, B.K. Fleischmann, H. Reuter, P. Sasse, H. Schunkert, J. Hescheler, Regulation of the Na+/Ca2+ exchanger (NCX) in the murine embryonic heart. Cardiovasc. Res. 75, 99–108 (2007)PubMedCrossRefGoogle Scholar
  44. U. Schlattner, M. Tokarska-Schlattner, T. Wallimann, Mitochondrial creatine kinase in human health and disease. Biochim. Biophys. Acta 1762, 164–180 (2006)PubMedCrossRefGoogle Scholar
  45. D.H. Schulze, M. Muqhal, W.J. Lederer, A.M. Ruknudin, Sodium/calcium exchanger (NCX1) macromolecular complex. J. Biol. Chem. 278, 28849–28855 (2003)PubMedCrossRefGoogle Scholar
  46. E.M. Schwarz, S. Benzer, Calx, a Na-Ca exchanger gene of Drosophila melanogaster. Proc. Natl. Acad. Sci. U. S. A. 94, 10249–10254 (1997)PubMedCrossRefGoogle Scholar
  47. M. Spindler, K. Meyer, H. Stromer, A. Leupold, E. Boehm, H. Wagner, S. Neubauer, Creatine kinase-deficient hearts exhibit increased susceptibility to ischemia-reperfusion injury and impaired calcium homeostasis. Am. J. Physiol. Heart Circ. Physiol. 287, H1039–H1045 (2004)PubMedCrossRefGoogle Scholar
  48. K. Steeghs, A. Benders, F. Oerlemans, A. de Haan, A. Heerschap, W. Ruitenbeek, C. Jost, J. van Deursen, B. Perryman, D. Pette, M. Bruckwilder, J. Koudijs, P. Jap, J. Veerkamp, B. Wieringa, Altered Ca2+ responses in muscles with combined mitochondrial and cytosolic creatine kinase deficiencies. Cell 89, 93–103 (1997)PubMedCrossRefGoogle Scholar
  49. A. Tokumura, M. Okuno, K. Fukuzawa, H. Houchi, K. Tsuchiya, M. Oka, Positive and negative controls by protein kinases of sodium-dependent Ca2+ efflux from cultured bovine adrenal chromaffin cells stimulated by lysophosphatidic acid. Biochim. Biophys. Acta 1389, 67–75 (1998)PubMedCrossRefGoogle Scholar
  50. K. Wakimoto, K. Kobayashi, O.M. Kuro, A. Yao, T. Iwamoto, N. Yanaka, S. Kita, A. Nishida, S. Azuma, Y. Toyoda, K. Omori, H. Imahie, T. Oka, S. Kudoh, O. Kohmoto, Y. Yazaki, M. Shigekawa, Y. Imai, Y. Nabeshima, I. Komuro, Targeted disruption of Na+/Ca2+ exchanger gene leads to cardiomyocyte apoptosis and defects in heartbeat. J. Biol. Chem. 275, 36991–36998 (2000)PubMedCrossRefGoogle Scholar
  51. T. Wallimann, M. Wyss, D. Brdiczka, K. Nicolay, H.M. Eppenberger, Intracellular compartmentation, structure and function of creatine kinase isoenzymes in tissues with high and fluctuating energy demands: the ‘phosphocreatine circuit’ for cellular energy homeostasis. Biochem. J. 281(Pt 1), 21–40 (1992)PubMedGoogle Scholar
  52. C.R. Weber, V. Piacentino 3rd, S.R. Houser, D.M. Bers, Dynamic regulation of sodium/calcium exchange function in human heart failure. Circulation 108, 2224–2229 (2003)PubMedCrossRefGoogle Scholar
  53. M.P. Wu, L.S. Kao, H.T. Liao, C.Y. Pan, Reverse mode Na+/Ca2+ exchangers trigger the release of Ca2+ from intracellular Ca2+ stores in cultured rat embryonic cortical neurons. Brain Res. 1201, 41–51 (2008)PubMedCrossRefGoogle Scholar
  54. W. Xu, H. Denison, C.C. Hale, C. Gatto, M.A. Milanick, Identification of critical positive charges in XIP, the Na/Ca exchange inhibitory peptide. Arch. Biochem. Biophys. 341, 273–279 (1997)PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  1. 1.Institute of Biochemistry and Molecular BiologyNational Yang-Ming UniversityTaipeiRepublic of China
  2. 2.Department of Life Sciences and Institute of Genome SciencesNational Yang-Ming UniversityTaipeiRepublic of China

Personalised recommendations