Publications on the project |
085 Calixarene Phosphonous Acids: Synthesis and Biological Activity |
Authors: | V. I. Kalchenko, S. O. Cherenok, S. O. Kosterin, E. V. Lugovskoy, S. V. Komisarenko, A. I. Vovk, V. Y. Tanchuk, L. A. Kononets & V. P. Kukhar | |
Summary: |
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Keywords: | Calixarenes, phosphonous acids, glutathione S-transferase, ATPases, fibrin polymerization, supramolecular complexes | |
Edition: | Phosphorus, Sulfur, and Silicon and the Related Elements, Special Issue: 19th International Conference on Phosphorus Chemistry (ICPC-2012) Rotterdam | | | 2013,
232-237,English |
085 Calixarene methylene bisphosphonic acids as promising effectors of biochemical processes. |
Authors: | S.V. Komisarenko, S.O. Kosterin, E.V. Lugovskoy, V.I. Kalchenko | |
Summary: | | |
Keywords: | | |
Edition: | In: BIOCHEMISTRY AND BIOTECHNOLOGY FOR MODERN MEDICINE. Edited by Prof. Serhiy Komisarenko. К.: Publishing House Moskalenko O.M. 2013, 704 p. | | | 2013,
293-327,English |
085 Modulators of transmembrane calcium exchange in myometrium mitochondria change their hydrodynamic diameter |
Authors: | N. V. Kandaurova, A. Ju. Chunikhin,
L. G. Babich, S. G. Shlykov, S. O. Kosterin
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Summary: | The influence of modulators of calcium exchange in mitochondria – oligomycin, Mg2+ and ruthenium red (RuR) – on the myometrium mitochondria size and granularity was studied in the work. The study of the mitochondria size was carried out using the photon correlation spectroscopy. It was shown that the average hydrodynamic diameter was 655 ± 14 nm (n = 5; control). The addition of oligomycin (1 μg/ml) – the inhibitor of АТР-synthase F0-component, increases the mitochondria average hydrodynamic diameter to 913 ± 75 nm (n = 5), that is by 39% more than the control. In the presence of RuR (10 μM) (Ca2+‑uniporter inhibitor) and Mg2+ (7 mM) the mitochondria average hydrodynamic diameter increases to 788 ± 28 and 788 ± 38 nm (n = 5) respectively, that is by 17% more than the control. Using flow cytometry it was shown, that oligomycin (1 μg/ml) causes the increase of side scattering of the mitochondria. Addition of RuR (10 μM) and Mg2+ (7 mM) does not lead to significant changes in side scattering of the mitochondria. So it was shown that oligomycin significantly increases mitochondria granularity, but Mg2+ and RuR have no influence on this parameter. | |
Keywords: | myometrium, isolated mitochondria, photon correlation spectroscopy, flow cytometry.
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Edition: | The Ukrainian Biochemical Journal, Kyiv, Ukraine | | | 2010,
52-57,Ukrainian |
085 Comparative investigation by spectrofluorimetry and flow cytometry of plasma and inner mitochondrial membranes polarisation in smooth muscle cell using potential-sensitive probe DiOC6 (3) |
Authors: | G.V. Danylovych, Yu.V. Danylovych, V.F. Gorchev | |
Summary: | Possibility of the use of flow cytometry and spectrofluorimetry analysis for investigation mitochondria and plasma membrane polarization in myometrium cell suspension using potential-sensitive probe 3,3′-dihexyloxacarbocyanine [DiOC6 (3)] has been demonstrated. The obtained results confirm the use of DiOC6 (3) for studying the influence of effectors on transmembrane potentials of intact cell compartments | |
Keywords: | transmembrane potential, potential-sensitive probe, flow cytometry, myometrium cell | |
Edition: | The Ukrainian Biochemical Journal | | | 2011,
99-105,Ukrainian |
085 Transmembrane Ca2+ exchange in depolarized rat myometrium mitochondria |
Authors: | L. G. Babich, S. G. Shlykov, N. V. Kandaurova, S. A. Kosterin
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Summary: | Polarization of the inner membrane is the key
factor in maintenance of the physiologically significant
cations accumulation, in particular Ca2+,
in the mitochondria. It has been well established
that mitochondria accumulate calcium through the
uniporter, driven by the mitochondrial membrane
potential. Nevertheless, it has been shown that
depolarized mitochondria also accumulate Ca2+.
The aim of this paper is to investigate free Ca level
in depolarized myometrium mitochondria. As we
have shown previously Ca2+ addition to the incubation
medium, that did not contain K-phosphate,
ATP and Mg2+, led to inner mitochondrial membrane
depolarization. Nevertheless Ca2+ addition to
such medium led to the concentration-dependent
accumulation of this cation in the matrix. RuR or
Mg addition to the incubation medium led to the
higher elevation of mitochondrial Ca2+ level in depolarized
mitochondria. Mitochondrial Ca2+ level
was not affected by 5 μM cyclosporine A. It was
suggested that Н+/Са2+ exchanger could provide
calcium accumulation in depolarized mitochondria.
The elevation of mitochondrial Ca2+ level
after addition of Mg2+ and RuR may be due to
inhibition of Ca2+ efflux through Ca2+ uniporter.
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Keywords: | isolated myometrium mitochondria; Ca2+; membrane potential; flow cytometry.
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Edition: | The Ukrainian Biochemical Journal, Kyiv, Ukraine | | | 2011,
56-62,Russian |
085 Comparative investigationof the effect of calix[4]arene C-99 and its analogs on Nа+,K+-ATPase activityof uterus myocite plasmamembrane |
Authors: | T.O. Veklich, A.A. Shkrabak, S.O. Cherenok, V.I. Kalchenko, S.O. Kosterin | |
Summary: | The aim of our investigation was to determine structural features of calix[4]arene C-99 which are important for its inhibition properties relative to Nа+,K+-ATPase of uterus myocite plasma membrane. Therefore we studied the effect of calix[4]arenes С-296, С-297, С-424, С-425, С-426, С-427, which are structurally similar to this inhibitor, on the mentioned enzyme activity. We have shown that calixarenes С-296 and С-297 which have two additional propoxy groups on the lower rim of macrocycle are less effective inhibitors of Nа+,K+-ATPase relative to calixarene C-99. Calixarenes С-425 and С-427 which have on the upper rim of macrocycle three and four phosponic residues, respectively, also inhibit Nа+,K+-ATPase activity less effectively as compared to calixarene C-99. Both calixarenes: С-424, which has only two carbonate residues on the upper rim, and С-426, which has on the upper rim ketomethilphosphonate residues instead of hydroxymethilphosphonate residues of calixarene C-99, do not affect Nа+,K+-ATPase activity. We have made respective conclusions concerning the role of certain chemical groups of calixarene C-99 during its interaction with Nа+,K+-ATPase. | |
Keywords: | Na+,K+-ATPase, Mg2+-ATPase, plasma membrane, smooth muscle cells, myometrium, kinetic properties of ATPase, calixarenes, aminophosphonic acids. | |
Edition: | Ukrainian biochemical journal | | | 2012,
49-57,Ukrainian |
085 Changes in polarization of myometrial cells plasma and internal mitochondrial membranes under calixarenes action as inhibitors of plasma membrane Na+,K+-ATPase |
Authors: | G.V. Danylovych, Yu.V. Danylovych, O.V. Kolomiets, S.O. Kosterin, R.V. Rodik, S.O. Cherenok, V.I. Kalchenko, A.Ju. Сhunikhin, V.F. Gorchev, S.A. Karakhim | |
Summary: | The influence of supramolecular macrocyclic compounds – calix[4]arenes C-97, C-99, C-107, which are ouabainomymetic high affinity inhibitors of Na+,K+-ATPase, on the polarization level of plasmic and mitochondrial membranes of rat uterine smooth muscle cells was investigated. The influence of these compounds on the myocytes characteristic size was studied . By using a confocal microscopy and specific for mitochondrial MitoTracker Orange CM-H2TMRos dye it was proved that the potential-sensitive fluorescent probe DiOC6(3) interacts with mitochondria. Artificial potential collapse of plasmic membrane in this case was modeled by myocytes preincubation with ouabain (1 mM). Further experiments performed using the method of flow cytometry with DiOC6(3) have shown that the compounds C-97, C-99 and C-107 at concentration 50-100 nM caused depolarization of the plasma membrane (at the level of 30% relative to control values) in conditions of artificial collapse of mitochondrial potential by myocytes preincubation in the presence of 5 mM of sodium azide. Under artificial sarcolemma depolarization by ouabain, calixarenes C-97, C-99 and C-107 at 100 nM concentrations caused a transient increase of mitochondrial membrane potential, that is 40% of the control level and lasted about 5 minutes. Calixarenes C-99 and C-107 caused a significant increase in fluorescence of myocytes in these conditions, which was confirmed by confocal microscopy too. It was proved by photon correlation spectroscopy method that the C-99 and C-107 caused an increase of characteristic size of myocytes | |
Keywords: | calix[4]arenes, plasma membrane, mitochondria, transmembrane potential, smooth muscle, uterus, photon correlation spectroscopy | |
Edition: | The Ukrainian Biochemical Journal | | | 2012,
37-48,Ukrainian |
085 Calix[4]arenes С_136 and С_137 Hyperpolarize Myometrium Mitochondria Membranes |
Authors: | L. G. Babich, , S. G. Shlykov, V. І. Boyko, M. A. Klyachina, S. A. Kosterin | |
Summary: | Calixarenes - supramolecular compounds interacting with bioactive molecules and ions that causes the changes in biochemical and biophysical processes. The aim of this work was to study the effects of calix[4]arenes С-136, С-137 and С-138 on the level of polarization of rat myometrium mitochondria membrane. Structure of synthesized calix[4]arene molecules was confirmed by the methods of 1Н NMR and infra-red spectroscopy. Calix[4]arenes С-136 and С-137 possess two chalcone amide moieties at the lower rim, while the calix[4]arene С-138 - only one. In case of calix[4]arenes С-136 and С-137 take place, accordingly, absence or presence of phenolic hydroxyl groups at the lower rim on the calix[4]arene skeleton. It was shown that calix[4]arenes С-136, С-137 and С-138 form micelles in a water medium and in the dimethylformamide (DМF). The irradiation of micelles with argon laser on flow cytometer results in appearance of autofluorescence. In the water medium calix[4]arene micelles interact with positively charged potential-sensitive fluorescent probe TMRM, that can testify to the presence of negative charge in these structures. However calix[4]arene micelles in DMF solution do not interact with TMRM. Mitochondrial membrane potential was measured using fluorescent dyes MTG and TMRM with confocal microscopy and fluorescent dye TMRM with flow cytometry. Experiments were conducted on myometrium cells in culture and on suspension of digitonin-permeabilized uterus myocytes. It was shown that a fluorescent signal was stable during time of experiment. Calix[4]arenes С-136 and С-137 (10 µМ) hyperpolarize mitochondria membranes. A maximal effect was 173%. At the same time calix[4]arene С-138 did not influence on mitochondria membrane potential. Connection comes into question between structural organization of investigated calix[4]arene molecules and their influence on polarization of mitochondria membrane. | |
Keywords: | calix[4]arenes, mitochondria membrane potential, myometrium | |
Edition: | Russian Journal of Bioorganic Chemistry | | | 2013,
728-735,Russian |
085 The сalix[4]arene C-107 is highly effective supramolecular inhibitor of the Na+,K+-ATPase of plasmatic membrane |
Authors: | O.V. Bevza, T.O. Veklich, O.A. Shkrabak, R.V. Rodik, V. I. Kalchenko, S.O. Kosterin | |
Summary: | The inhibition of the Na+,K+-АТРase activ-ity of the myometrium cell plasma membranes with calixarene С-107 (5,17-diamino(2-pyridyl)methylphosphono-11,23-di-tret-butyl-26,28-dihy-droxy-25,27-dipropoxycalix[4]arene) was investigated. It has been shown that calixarene С-107 reduced the Na+,K+-АТРase activity more efficiently than ouabain did, while it did not practically influence the «basal» Mg2+-АТРase activity of the same membrane. The magnitude of the cofficient of inhibition I0.5 was 33 ± 4 nМ, Hill coefficient was 0.38 ± 0.06. The model calixa rene C-150 – the calixarene «scaffold» (26,28-dihydroxy-25,27-dipropoxycalix[4]arene), and the model compound М-3 (4-hydroxyaniline(2-pyridine)methylphosphonic acid) – a fragment of the calixarene С-107, had practically no influence on the enzymatic activity of Na+,K+-АТРase and Mg2+-АТРаse. We carried out the computer simulation of interaction of calixarenes C-107 and the mentioned model compound with ligand binding sites of the Na+,K+-АТРase of plasma membrane and structure foundation of their intermolecular interaction was found out. The participation of hydrogen, hydrophobic, electrostatic and π-π(stacking) interaction between calixarene and enzyme aminoacid residues, some of which are located near the active center of Na+,K+-АТРase, was discussed | |
Keywords: | Na+,K+-ATPase, plasma membrane, smooth muscle cells, myometrium, calix[4]arene C-107, computer simulation, docking. | |
Edition: | Ukrainian Biochemical Journal | | | 2013,
5-19,Ukrainian |
085 Kinetic regularities of calixarene C-90 action on the myometrial plasma membrane Ca2+,Mg2+-ATPase activity and on the Ca2+ concentration in unexcited сells of the myometrium |
Authors: | T. O. Veklich, A. A. Shkrabak, Yu. Yu. Mazur, R. V. Rodik, V. I. Boyko, V. I. Kalchenko, S. O. Kosterin | |
Summary: | Plasma membrane Са2+,Mg2+-ATPase is an important element of general myometrium tonus control mechanism, which also makes a contribution to muscle tension relaxation after its contraction. Expiriments were done on the myometrial cell plasma membrane suspension, which was treated with 0.1% digitonin solution. The authors have investigated the inhibitory action of calix[4]arene C-90 (5,11,17,23-tetra(trifluor) methyl(phenylsulphonylimino)-methylamino25,26,27,28-tetra propoxi-calix[4]arene) on the Са2+,Mg2+-ATPase activity (the magnitude of і0.5 was 20.2 ± 0.5 mkM). The inhibitory action of calix[4]arene C-90 on the activity of Са2+,Mg2+-ATPase is explained as cooperative action of four trifluormethyl(phenylsulfonylimino)methylamino groups that are spatially oriented on the calix[4]arene base rather than with the action of tetraphenol macrocycle or separate pharmacophore sulphonilamidin groups. Considering established kinetic pattern of calix[4]arene C-90 inhibitory action on the plasma membrane Са2+,Mg2+-ATPase activity, stationary kinetical model of basal calcium concentration control in unexcited uterus myocytes was developed. It is assumed that obtained results may be promising for creation of new generation («supramolecular») pharmacological agent - uterus basal tonus stimulator – on the base of calix[4] arene C-90. | |
Keywords: | Ca2+,Mg2+-ATPase, plasma membrane, smooth muscle cells, myometrium, enzymatic hydrolysis of ATP, kinetic properties of ATPase, calix[4]arenes. | |
Edition: | Ukrainian biochemical journal | | | 2013,
20-29,Ukrainian |
085 Calix[4]arene С-90 selectively inhibits Ca2+,Mg2+-АТРаse of myometrium cell plasma membrane |
Authors: | Veklich T.O., Shkrabak A.A., Slinchenko N.N., Mazur I.I., Rodik R.V., Boyko V.I., Kalchenko V.I., Kosterin S.O | |
Summary: | The supramolecular compound calix[4]arene C-90 (5,11,17,23-tetra(trifluoro)methyl(phenylsulfonylimino)methylamino-25,26,27,28-tetrapropoxycalix[4]arene) is shown to efficiently inhibit the ATP hydrolase activity of
Ca2+,Mg2+ATPase in the myometrium cell plasma membrane fraction and also in a preparation of the purified enzyme solubilized from this subcellular fraction. The inhibition coefficient I0.5 values were 20.2 ± 0.5 and 58.5 ± 6.4 μM for the membrane fraction and the solubilized enzyme, respectively. The inhibitory effect of calix[4]arene C-90 was selective comparatively to other ATPases localized in the plasma membrane: calix[4]arene C-90 did not influence the activities of Na+,K+-ATPase and “basal” Mg2+-ATPase. The inhibitory effect of calix[4]arene C-90 on the Ca2+,Mg2+ATPase activity was associated with the cooperative action of four trifluoromethylphenyl sulfonylimine (sulfonylamidine) groups oriented similarly on the upper rim of the calix[4]arene macrocycle (the calix[4]arene “bowl”). The experimental findings seem to be of importance for studies, using calix[4]arene C-90, of membrane mechanisms of regulation of calcium homeostasis in smooth muscle cells and also for investigation of the participation of the plasma membrane Ca2+pump in control of electro and pharmacomechanical coupling in myocytes.
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Keywords: | Ca2+,Mg2+-ATPase, plasma membrane calcium pump, smooth muscle cells, myometrium, calix[4]arenes, sulfonylamidines | |
Edition: | Biochemistry (M) | | | 2014,
417-424,Russian |
085 The derivatives of imidazo[1,2-a]azepine as modifiers of myometrium contractile activity |
Authors: | N. A. Mochort, I. V. Gerashchenko, A. A. Shkrabak, Yu. Yu. Mazur, T. O. Veklich | |
Summary: | Preterm birth is a major contributor to perinatal mortality and morbidity worldwide. Tocolytic agents are drugs used to inhibit uterine contractions and to prevention preterm delivery. In this study we investigated the effects of the derivatives of imidazo[1,2-a]azepinium on oxytocin-induced contractions in the rat uterus in vitro and ATPases activity in myometrium cell’s plasma membrane. Contractile activity was studied isometrically and analysis of contractions was performed by the calculation of the basal tone, amplitude, interval between cycles of reduction, the phase of contraction and relaxation. Spontaneously active myometrium was activated with oxytocin which was added to the tissue bath solution. After oxytocin administration the derivatives of imidazo[1,2-a]azepinium (0,1; 1; 10 µM) were added to the bath solution too. We investigated the activity of Ca2+,Mg2+-ATPase, Na+,K+-ATPase, Mg2+-ATPase, Ca2+-ATPase on the myometrial cell plasma membrane suspension. Student`s t-test was used for statistical analysis. It has been found out that observed increasing in time parameters and tone depend on oxymethyl and oxyethyl residues in structures studied. It was suggested that remarkable increasing in time parameters depends on presence of oxygen and nitrogen atoms which can form hydrogen bonds with hydrogen in chemical structures of derivatives of imidazo[1,2-a]azepinium. The date obtained shown that derivatives of imidazo[1,2-a]azepinium IFT-176, IFT-208 activate Ca2+,Mg2+-ATPase and inhibit Na+,K+-ATPase. The inhibition uterine horn’s muscle stripes contractions caused by oxytocine may be due to Ca2+,Mg2+- ATPase activation. It is bloks one biochemical pathway oxytocine influence on calcium homeostasis of myometrium. | |
Keywords: | imidazo[1,2-a]azepinium derivatives, Ca2+,Mg2+-ATPase, Na+,K+-ATPase, Mg2+-ATPase, Ca2+-ATPase, plasma membrane, smooth muscle cells, myometrium, oxytocine | |
Edition: | Pharmacology and drug toxicology | | | 2014,
65-72,Ukrainian |
085 Kinetics of inhibitory effect of calix[4]arene C-90 on activity of transporting plasma membrane Cа2+, Mg2+-ATPase of smooth muscle cells |
Authors: | T. O. Veklich, A. A. Shkrabak, Yu. Yu. Mazur, R. V. Rodik, V. I. Kalchenko, S. O. Kosterin | |
Summary: | In experiments on the suspension of myometrium cell plasma membrane, processed by 0.1% digitonin, the inhibitory action of calix[4]arene C-90 (5,11,17,23-tetra(threeftor)methyl(phenilsulphonilimino)-methylamino-25,26, 27,28-tetrapropoxy-calix[4]arene) on the activi¬ty of Ca2+,Mg2+-ATPase was investigated. The authors also examined the influence of calix[4]arene in different concentration on affinity of enzyme (Ca2+,Mg2+-ATPase) for the ATP and ions of Mg and Ca, and its influence on cooperative effect and maximum velocity of ATP hydrolysis. It is shown that calix[4]arene does not influence the affinity of Ca2+,Mg2+-ATPase for the ATP, which means that these two compounds have different binding centers¬. Also calix[4]arene has no influence on affinity and cooperative effect of Ca ions, if it is used in concentration lower than 50 µM. Calix[4]arene slightly increases coefficient of Ca2+,Mg2+-ATPase activation by magnesium chloride. In all three cases, where ATP, Mg and Ca ions are used to test the impact of calix[4]arene, maximum velocity of ATP hydrolysis significantly decreases. All these results clarify that calix[4]arene implements its inhibitory action through mechanism of uncompetitive inhibition of Ca2+,Mg2+-ATPase activity. | |
Keywords: | Ca2+,Mg2+-ATPase, plasma membrane, smooth muscle cells, myometrium, enzymatic hydrolysis of ATP, kinetic properties of ATPase¬, calix[4]arenes. | |
Edition: | Ukrainian biochemical journal | | | 2014,
37-46,Ukrainian |
085 Plasma membrane Ca2+,Mg2+-ATPase activity is selectively suppressed by calIx[4]arene C-90 |
Authors: | Т. Veklich, О. Shkrabak, Iu. Mazur | |
Summary: | In experiments, which were performed on myometrium plasma membrane suspension processed by 0,1% digitonin, effect of calix[4]arene C-90 (5,11,17,23-tetra(threephtor)methyl(phenilsulphonilimino)-methylamino-25,26,27,28-tetra propoxi-calix[4]arene) on Ca2+,Mg2+-ATPase activity was investigated. Calix[4]arene C-90 effectively inhibited Ca2+,Mg2+-ATPase activity (І0,5 was 20,2±0,5 µM). Confocal microscopy measurement showed that C-90 (20 µM) temporarily increase intracellular Са2+ concentration. Obtained results allows to assume that calix[4]arene may be useful as a basis for creation of new pharmacological agent for basal uterus tonus stimulation. | |
Keywords: | Ca2+,Mg2+-ATPase, plasma membrane, myometrium, calix[4]arene | |
Edition: | Visnyk of the Lviv University | | | 2014,
337-347,Ukrainian |
085 Modulation of myometrium mitochondrial membrane |
Authors: | S. G. Shlykov, L. G. Babich, M. E. Yevtushenko, S. O. Karakhim,
S. O. Kosterin
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Summary: | Influence of calmodulin antagonists on mitochondrial membrane potential was investigated using a flow cytometry method, confocal microscopy and fluorescent potential-sensitive probes TMRM and MTG. Influence of different concentrations of calmodulin antagonists on mitochondrial membrane potential was studied using flow cytometry method
and a fraction of myometrium mitochondria of unpregnant
rats. It was shown that 1-10 μМ calmidazolium gradually reduced mitochondria membrane potential. At the same time 10-100 μМ trifluoperazine influenced as follows: 10 μМ – increased polarization, while 100 μМ – caused almost complete depolarization of mitochondrial membranes. In experiments
which were conducted with the use of confocal microscopy method and myometrium cells it was shown, that MTG addition to the incubation medium led to the appearance of fluorescence signal in a green range. Addition of the second probe (ТМRM)
resulted in the appearance of fluorescent signal in a red range. Mitochondrial membrane depolarization by 1μМ СССР or 10 mМ NaN3 was accompanied by the decline of “red” fluorescence
intensity, “green” fluorescence was kept. The 10-15 minute incubation of myometrium cells in the presence 10 μМ calmidazolium or 100 μМ trifluoperazine was accompanied
by almost complete decrease of the TMRM fluorescent
signal. Thus, with the use of potential-sensitive fluorescent probes TMRM and MTG it was shown, that calmodulin antagonists modulate mitochondrial membrane potential of myometrium cells.
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Keywords: | flow cytometry, confocal fluorescence microscopy, isolated mitochondria, smooth muscles cells, mitochondrial membrane potential, calmidazolium, trifluoperazine.
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Edition: | The Ukrainian Biochemical Journal | | | 2014,
29-41,Ukrainian |
085 Ca2+/H+-exchange in myometrium mitochondria |
Authors: | O.V. Kolomiets, Yu.V. Danylovych, H.V. Danylovych, S.O. Kosterin | |
Summary: | Using the fluorescent probe Fluo-4 AM the authors have identified Na+-independent Ca2+/H+-exchange in isolated mitochondria of rat myometrium and studied its individual properties. Formation of directional protons gradient in the matrix of mitochondria causes antyporte release of Ca2+, which has been previously accumulated in energetic processes (in the presence of Mg-ATP and succinate). The functioning of Ca2+/H+-exchange depends on the proton gradient and is characterized by reversibility, in case of extramitochondria environment alkalization the additional accumulation of Ca2+ by organelles is recorded. Monovalent cations gradients (Na+, K+, Li+) do not cause the release of Ca2+ from mitochondria. Rate of Ca2+/H+-exchange is growing in terms of increasing ΔpH on the mitochondria membrane and kinetics of ΔpH-induced Ca2+ release from the matrix corresponds to the laws of first order reaction. Research of Ca2+/H+-exchange some properties in the myometrium mitochondria showed that the above transport process is of electrogenic nature, perhaps it is done in a 1 : 1 stechiometry (Hill coefficient on H+ close to 1) and is able to adjust matrix Ca2+ concentration under physiological conditions (pH activation of about 6,9). Thus, in the inner membrane of the myometrium mitochondria the available system of the secondary active Ca2+-transport from the matrix of these organelles to myoplasm and the functioning of Ca2+/H+-exchanger may underlie this process | |
Keywords: | mitochondria, Ca2+/H+-exchange, smooth muscle, myometrium, Letm1 | |
Edition: | The Ukrainian Biochemical Journal | | | 2014,
41-48,Ukrainian |
085 Regulation of the mitochondrial ATP-sensitive potassium channel in rat uterus cells by ROS |
Authors: | O.B. Vadzyuk, Yu.Yu. Mazur, S.O. Kosterin | |
Summary: | In previous study we demonstrated the presence of ATP-sensitive potassium current in the inner mitochondrial membrane, which was sensitive to diazoxide and glybenclamide, in mitochondria isolated from rat uterus. This current was supposed to be operated by mitochondrial ATP-sensitive potassium channel (mitoKATP). Regulation of the mitoKATP in uterus cells is not studied well enough yet.
It is well known that the reactive oxygen species (ROS) can play a dual role. They can damage cells in high concentrations, but they can also act as messengers in cellular signaling, mediating survival of cells under stress conditions. ROS are known to activate mitoKATP during the oxidative stress in brain and heart, conferring the protection of cells. The present study examined whether ROS mediate the mitoKATP activation in myometrium cells.
Oxidative stress was induced by rotenone. ROS generation was measured by 2',7'- dichlorofluorescin diacetate. The massive induction of ROS production was demonstrated in the presence of rotenone. Hyperpolarization of the mitochondrial membrane was also detected with the use of the potential-sensitive dye DiOC6 (3,3'-dihexyloxacarbocyanine iodide). Diazoxide, a selective activator of mitoKATP, depolarized mitochondrial membrane either under oxidative stress or under normal conditions, while mitoKATP blocker glybenclamide effectively restored mitochondrial potential in rat myocytes. Estimated <K1/2> value for diazoxide to mitoKATP under normoxia was four times higher than under oxidative stress conditions: 5.01 ± 1.47 × 10-6 М and 1.24 ± 0.21 × 10-6 М respectively. The ROS scavenger N-acetylcysteine (NAC) successfully eliminates depolarization of mitochondrial membrane by diazoxide under oxidative stress. These results suggest that elimination of ROS by NAC prevents the activation of mitoKATP under oxidative stress. Taking into account the higher affinity of diazoxide to mitoKATP under stress conditions than under normoxia, we conclude that the oxidative stress conditions are more favorable than normoxia for the activation of mitoKATP. Thus we hypothesize that the ROS activate the mitoKATP in myocytes.
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Keywords: | mitochondrial ATP-sensitive K+-channel, diazoxide, reactive oxygen species
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Edition: | Ukrainian Biochemical Journal | | | 2011,
48-57,Ukrainian |
085 Structural and functional basis of the intermolecular interaction of calix[4]arene C-97 with myosin subfragment-1 of myometrium |
Authors: | R. D. Labyntseva, A. A. Bevza, О. V. Bevza, S. O. Cherenok,V. I. Kalchenko, S. O. Kosterin | |
Summary: | Calix[4]arenas C-97 (code is shown) - this macrocyclic substans which have an lipophilic intramolecular higly-structured cavity formed by four aromatic macrocycles, one with which on the upper rim modified by methylene bisphosphonic group. It was shown that calix[4]arenas C-97 (100 µM) efficiently inhibits ATPase activity of myosin subfragment-1 from pig myometrium, the value of the constant inhibition І0,5 being 83 ± 7 µM. At the same time, this compound at 100 µM concentration significantly increases an effective hydrodynamic diameter of myosin subfragment-1, that may be indicative of intermolecular complexation between the calix[4]arene and the myosin head. We used computer simulation (docking, molecular dynamics, involving the Grid) to clarify structural basis of the intermolecular interaction of calix[4]arene C-97 with myosin subfragment-1 of the myometrium. According to received data about the dynamics of entrenching of calix[4]arene C-97 in the ligand-binding center of the myosin head there are several hydrophobic and electrostatic interactions between calix[4]arene C-97 and amino acid residues of myosin subfragment-1, some of them located near the active site of the ATPase. | |
Keywords: | ATPase, subfragment-1 of myosin, calix[4]arene C-97, smooth muscle, uterus.
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Edition: | The Ukrainian Biochemical Journal | | | 2012,
34 – 44.,Ukrainian |
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085 5. Nanobiotechnology Purpose: Expected results:Issue of new types of products: methods, theories Stage 1: Stage 2: Stage 3: Stage 4: Stage 5:
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