Kinetic regularities of thiacalix[4]arene C-1087 inhibitory effect on the activity of Mg(2+)-dependent Ca(2+)-transporting ATP hydrolase in the plasma membrane of smooth muscle cells

Т. О. Veklich, О. V. Bevza, О. V. Maliuk, S. Kosterin, R. Rodik, S. Vyshnevskyi, V. І. Kalchenko
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Abstract

The experiments with the suspension of plasma membranes of myometrium cells, treated with 0.1% digitonin solution, were used to study kinetic regularities of the inhibitory effect of tetra-N-phenylsulfonyl trifluoroacetamidine-thiacalixarene (С-1087) on the activity of Са2+,Mg2+-ATPase. The studies demonstrated the impact of C-1087 on the cumulative effect and the maximal velocity of ATP hydrolysis. No effect of С-1087 on the affinity between Са2+,Mg2+-ATPase, and АТР, affinity and cumulative effect of Ca ions and activation coefficient for Mg ions was revealed. A considerable decrease in the maximal velocity of ATP hydrolysis evidenced a complete non-competitive mechanism of inhibiting Са2+,Mg2+-АТРase activity with thiacalix[4]arene С-1087. Computer simulation demonstrated that thiacalix[4]arene С-1087 inhibiting effect on Са2+,Mg2+-ATPase may be conditioned by the cumulative effect of four spatially oriented N-sulfonylamidine groups on the upper rim of its macrocyclic platform.
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硫杂六[4]炔 C-1087 对平滑肌细胞质膜中依赖 Mg(2+)Ca(2+)-transporting ATP 水解酶活性的抑制作用的动力学规律性
用 0.1% 地西妥宁溶液处理的子宫肌细胞浆膜悬浮液进行实验,研究了四-N-苯磺酰基三氟乙脒-硫杂堇菜烯(С-1087)对Са2+,Mg2+-ATP 酶活性的抑制作用的动力学规律。研究表明,C-1087 对 ATP 水解的累积效应和最大速度有影响。С-1087对Са2+,Mg2+-ATPase和АТР之间的亲和力、钙离子的亲和力和累积效应以及镁离子的活化系数没有影响。ATP 水解的最大速度显著降低,这证明硫杂[4]炔С-1087 对Са2+,Mg2+-АТР酶活性的抑制机制是完全非竞争性的。计算机模拟表明,硫杂[4]炔С-1087 对Са2+,Mg2+-АТР酶的抑制作用可能受其大环平台上缘四个空间取向的 N-磺酰基脒基团累积效应的制约。
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