Ukr.Biochem.J. 2018; Volume 90, Issue 1, Jan-Feb, pp. 25-33


Cаlіx[4]аrene С-956 is effective inhibitor of Н(+)-Сa(2+)-exchanger in smooth muscle mitochondria

G. V. Danylovych1, О. V. Kоlomiets1, Yu. V. Danylovych1, R. V. Rodik2, V. I. Kаlchenko2, S. О. Kоsterin1

1Palladin Institute of Biochemistry, National Academy of Science of Ukraine, Kyiv;
2Institute of Organic Chemistry, National Academy of Science of Ukraine, Kyiv

It was shown that calix[4]arene C-956 exhibited a pronounced concentration-dependent (10-100 μM) inhibitory effect on the H+-Ca2+-exchanger of the inner mitochondrial membrane of rat uterine myocytes (Ki 35.1 ± 7.9 μM). The inhibitory effect of calix[4]arene C-956 was accompanied by a decrease in the initial rate (V0) and an increase in the magnitude of the characteristic time (τ1/2) of the ΔрН-induced Са2+ release. At the same time, it did not affect the potential-dependent accumulation of Ca2+ in mitochondria. Thus, the action of calix[4]arene C-956 might be directed on increasing the concentration of Ca ions in the mitochondrial matrix. The calculation of basic kinetic parameters of the Ca2+ transport from isolated organelles (in the case of its non-zero stationary level), based on changes in fluorescence of Ca2+-sensitive dye Fluo-4 AM in mitochondria was performed. The proposed approach can be used for the kinetic analysis of the exponential decrease of the fluorescence response of any probes under the same experimental conditions.

Keywords: , , , ,


  1. Kostyuk PG, Kostyuk OP, Lukyanets EA. Intracellular calcium signaling: structures and functions. Kyiv: Naukova Dumka, 2010. 175 p. (In Ukrainian).
  2. Bernardi P, Rasola A. Calcium and cell death: the mitochondrial connection. Subcell Biochem. 2007;45:481-506. PubMed, CrossRef
  3. Graier WF, Frieden M, Malli R. Mitochondria and Ca2+ signaling: old quests, new functions. Eur J Physiol. 2007; 455(3): 375-96. PubMed, PubMedCentral, CrossRef
  4. Mitochondrial signaling in health and disease. Eds. S. Orrenius, L. Packer, E. Cadenas. N.Y.: CRC Press, 2012. 493 p.  CrossRef, CrossRef
  5. Chalmers S, Nicholls DG. The relationship between free and total calcium concentrations in the matrix of liver and brain mitochondria. J Biol Chem. 2003 May 23;278(21):19062-70.  PubMed, CrossRef
  6. Takeuchi A, Kim B, Matsuoka S. The destiny of Ca(2+) released by mitochondria. J Physiol Sci. 2015 Jan;65(1):11-24.  PubMed, PubMedCentral, CrossRef
  7. Giuliani M, Morbioli I, Sansone F, Casnati A. Moulding calixarenes for biomacromolecule targeting. Chem Commun (Camb). 2015 Sep 28;51(75):14140-59.
    PubMed, CrossRef
  8. Danylovych GV, Danylovych YuV, Kolomiets OV, Kosterin SO, Rodik RV, Cherenok SO, Kalchenko VI, Сhunikhin AJu, Gorchev VF, Karakhim SA. Changes in polarization of myometrial cells plasma and internal mitochondrial membranes under calixarenes action as inhibitors of plasma membrane Na+, K+-ATPase. Ukr Biokhim Zhurn. 2012 Nov-Dec;84(6):37-48. (In Ukrainian). PubMed
  9. Danylovych НV, Danylovych YuV, Rodik RV, Kalchenko VI, Chunikhin AJu. Calix[4]arenes as modulators of energy-dependent Ca2+-accumulation and functioning of the electron transport chain in smooth muscle mitochondria. Fisiol Zh. 2016;62(5):27-36. (In Ukrainian). CrossRef
  10. Kolomiets ОV, Danylovych YuV, Danylovych НV, Kosterin SО. Ways and mechanisms of  transmembrane exchange of Ca2+ in mitochondria. Fisiol Zh. 2017; 63(4): 87-104. (In Ukrainian).  CrossRef
  11. Csordás G, Várnai P, Golenár T, Sheu SS, Hajnóczky G. Calcium transport across the inner mitochondrial membrane: molecular mechanisms and pharmacology. Mol Cell Endocrinol. 2012 Apr 28;353(1-2):109-13.  PubMed, PubMedCentral, CrossRef
  12. Yoshino M1, Wang SY, Kao CY. Sodium and calcium inward currents in freshly dissociated smooth myocytes of rat uterus. J Gen Physiol. 1997 Nov;110(5):565-77. PubMed, PubMedCentral, CrossRef
  13. Seda M, Pinto FM, Wray S, Cintado CG, Noheda P, Buschmann H, Candenas L. Functional and molecular characterization of voltage-gated sodium channels in uteri from nonpregnant rats. Biol Reprod. 2007 Nov;77(5):855-63. PubMed, CrossRef
  14. Kosterin SA. The possible H+-dependent functional connection between cell membrane and mitochondria in smooth muscle cells. Ukr Biokhim Zhurn. 1998 Nov-Dec;70(6):152-60. (In Russian).PubMed
  15. Vovkanich LS, Dubitsky LO. Kinetic properties of Н+-stimulated exit of Са2+ from liver mitochondria. Exp Clin Physiol Biochem. 2001; 3(15): 34-37. (In Ukrainian).
  16. Jiang D, Zhao L, Clapham DE. Genome-wide RNAi screen identifies Letm1 as a mitochondrial Ca2+/H+ antiporter. Science. 2009 Oct 2;326(5949):144-7.
    PubMed, PubMedCentral, CrossRef
  17. Santo-Domingo J, Wiederkehr A, De Marchi U. Modulation of the matrix redox signaling by mitochondrial Ca2+. World J Biol Chem. 2015 Nov 26;6(4):310-23.  PubMed, PubMedCentral, CrossRef
  18. Kolomiets OV, Danylovych YuV, Danylovych HV, Kosterin SO. Ca2+/H+-exchange in myometrium mitochondria. Ukr Biochem J. 2014 May-Jun;86(3):41-8. (In Ukrainian). PubMed, CrossRef
  19. Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248-54. PubMed, CrossRef
  20. Applied Statistics in Biological Research. Basic concepts, related problems and common methods / Dr. Karin Schmid. CSF vienna BIOCENTER. 2014/
  21. Veklich TO. The inhibitory influence of calix[4]Arene of C-90 on the activity of Ca2+,Mg2+-ATPases in plasma membrane and sarcoplasmic reticulum in myometrium cells. Ukr Biochem J. 2016 Mar-Apr;88(2):5-15. PubMed, CrossRef
  22. Veklich T, Shkrabak O, Nikonishyna Yu. Calix[4]aren C-956 selectively inhibits Ca2+,Mg2+-ATPases of plasma membrane. The 4th international scientific conference current problems of biochemistry and cell biology. Program and abstracts. 5-6 October, 2017. Dnipro, Ukrain: 233-234. (In Ukrainian).
  23. Babich LG, Fomin VP, Kosterin SA. Effect of the membrane potential on the Mg2+,ATP-dependent transport of Ca2+ across smooth muscle sarcolemma. Biokhimiia. 1990 Oct;55(10):1890-901. (In Russian). PubMed

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