CALIX[4]ARENES AS MODULATORS OF ENERGY-DEPENDENT CА2+- ACCUMULATION AND FUNCTIONING OF THE ELECTRON TRANSPORT CHAIN IN SMOOTH MUSCLE MITOCHONDRIA
Н.V. Danylovych1, Yu.V. Danylovych1, R.V. Rodik2, V.I. Kalchenko2, A.Ju. Chunikhin1
- Palladin Institute of Biochemistry, National Academy of
Sciences of Ukraine, Kyiv;
- Institute of Organic Chemistry,
National Academy of Sciences of Ukraine, Kyiv;
DOI: https://doi.org/10.15407/fz62.05.027
![](/images/cite.png)
Abstract
The influence of supramolecular macrocyclic compounds
сalix[4]arenes (C-97, C-99, C-107) at a concentration of
100 nM in the process of energy-dependent Ca2+-transport
in isolated mitochondria of smooth muscle, as well as
autofluorescence mitochondrial coenzyme NADH, FAD and
hydrodynamic diameter of these organelles was investigated.
Using Ca2+-sensitive fluorescent dye Fluo-4 AM it was shown
that the selected calix[4]arenes can suppress energy-dependent
accumulation of Ca2+ by mitochondria. Accumulation of
Ca2+ (80 µM in the medium) accompanied by the growth of
the fluorescent probe response from a conventional unit to a
value of 1,57±0,04 (n=5). Calix[4]arenes C-97, C-99, C-107
falls fluorescent signal below the 0,88±0,08, 0,92±0,08 and
0,78±0,04 respectively. Thus, the selected calix[4]arenas lead
to release of previously accumulated Ca2+ from mitochondria.
Under the influence of C-97 and C-99 fluorescent signal from
NADH reduced to -0,11±0,02 and -0,12±0,02, respectively,
in relation to the reference value - -0,05±0,01 (n=5). Analysis
of fluorescence response NADH and FAD in a suspension
of isolated mitochondria suggests that the effects of test
compounds on the functional activity of the electron transport
chain is associated with the initial stimulation of its I-th
complex and subsequent inhibition of Ca2+-dependent NADcontaining
Krebs cycle dehydrogenases. Along with this, the
use of photon correlation spectroscopy to assess changes in
the volume of mitochondria (their hydrodynamic diameter)
under the action of selected calix[4]arenes has shown that
interference with the electron transport chain leads to changes
in the osmotic balance between the matrix of the mitochondria
and the external environment. The result is the growth of
isolated organelles volume. In particular, the hydrodynamic
diameter of mitochondria increased by 22±6 % and 34±8 %
(n=5) in presence of C-97 or C-99. The conclusion was done
about the advisability of further studies of the calyx[4]arenes
effect on smooth muscle Ca2+-homeostase and mitochondrial
bioenergetics in order to find effective modifiers of their functional
activity.
Keywords:
mitochondria; electron transport chain; Ca2+- transport; smooth muscle.
References
- Kostyuk PG, Kostyuk OP, Lukyanets EA. Intracellular calcium signaling: structures and functions. Kiev: Naukova dumka, 2010. [Ukrainian].
- Santo-Domingo J, Demaurex N Calcium uptake mechanisms of mitochondria. Biochim Biophys Acta. 2010 Jun-Jul;1797(6-7):907-12
- Pan S1, Ryu SY, Sheu SS. Distinctive characteristics and functions of multiple mitochondrial Ca2+ influx mechanisms. Sci China Life Sci. 2011 Aug;54(8):763-9
- Wang HW, Wei YH, Guo HW. Reduced nicotinamide adenine dinucleotide (NADH) fluorescence for detection of cell death. Anticancer Agent Med Chem. 2009; 9(9):1012-7
CrossRef
- Heikal AA. Intracellular coenzymes as natural biomarkers for metabolic activities and mitochondrial anomalies. Biomark Med. 2010;4(2):241-63.
CrossRef
PubMed PubMedCentral
- Gellerich FN, Gizatullina Z, Trumbeckaite S, Nguyen HP, Pallas T, Arandarcikaite O, Vielhaber S, Seppet E, Striggow F. The regulation of OXPHOS by extramitochondrial calcium. Biochim Biophys Acta. 2010; 1797(6-7):1018-27.
- Giuliani M, Morbioli I, Sansone F, Casnati A moulding calixarenes for biomacromolecule targeting. Chem. Commun., 2015, DOI: 10.1039/C5CC05204A.
CrossRef
- Danylovych HV, Danylovych IuV, Kolomiiets' OV, Kosterin SO, Rodik RV, Cherenok SO, Kal'chenko VI, Chunikhin OIu, Horchev VF, Karakhim SO. Changes in polarization of myometrial cells plasma and internal mitochondrial membranes under calixarenes action as inhibitors of plasma membrane Na+, K+-ATPase. Ukr Biokhim Zh (1999). 2012 Nov-Dec;84(6):37-48. [Ukrainian].
- Kosterin SA, Bratkova NF, Rursky MD. The role of sarcolemma and mitochondria in Ca2+-dependent control of myometrium relaxation. Biochemistry (Moscow). 1985; 50(8):1350-1361. [Russian].
- 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; 72:248-54.
CrossRef
- Kolomiets' OV, Danylovych IuV, Danylovych HV, Kosterin SO. Ca2+ accumulation study in isolated smooth muscle mitochondria using Fluo-4 AM. Ukr Biochem J. 2013 Jul-Aug;85(4):30-9 [Ukrainian].
- Bailay NTJ. Statistical methods in biology. Great Britain: Cambridge University Press; 1995.
- Malli R, Graier WF. Mitochondrial Ca2+ channels: Great unknowns with important functions.FEBS Lett. 2010; 584:1942-47.
CrossRef
PubMed PubMedCentral
- Wei AC, Liu T, Cortassa S, Winslow RL, O'Rourke B. Mitochondrial Ca2+ influx and efflux rates in guinea pig cardiac mitochondria: low and high affinity effects of cyclosporine A. Biochim Biophys Acta. 2011;1813:1373-81.
CrossRef
PubMed PubMedCentral
- Kolomiets' OV, Danylovych IuV, Danylovych GV. H+-Ca2+-exchanger in the myometrium mitochondria: modulation of exogenous and endogenous compounds. Fisiol. Zh. 2014;60(5):33-42. [Ukrainian].
- Veklich TO, Koshechkova NS, Rodik RV, Boĭko VI, Vorobets' ZD, Kosterin SO. Comparative study of calixarene effect on Na+, K+ -ATPase activity in plasma membrane of contractile and mobile cell. Ukr Biokhim Zh (1999). 2007;79(3):19-28. [Ukrainian].
- Agarwal B, Dash RK, Stowe DF, Bosnjak ZJ, Camara AK. Isoflurane modulates cardiac mitochondrial bioenergetics by selectively attenuating respiratory complexes. Biochim Biophys Acta. 2014; 1837(3): 354–365.
CrossRef
PubMed PubMedCentral
|