Slinchenko N N, Taran T T, Kosterin S A
A.V.Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv.
Ukr Biokhim Zh (1978). 1997 Jul-Aug;69(4):9-17.
Some organic solvents (2-10%) have been comparatively studied for their effect on purified transporting Ca2+, Mg(2+)-ATPase, solubilized from the plasma membrane of smooth muscle cells and on actomyosine ATPase of the smooth muscle. The inhibiting effect of solvents on the initial maximum specific activity of Ca2+, Mg(2+)-ATPase corresponds to the sequence dioxane > acetone > ethanol > dimethyl sulfoxide (DMSO). Like the case with Ca2+, Mg(2+)-ATPase, dioxane inhibits actomyosine ATPase; acetone, ethanol and DMSO stimulate ATP-hydrolase reaction which is catalyzed by the complex of contractile proteins. It is proved that the effect of the decrease of ATPase activity with decrease of incubation medium polarity is exceptionally determined by the value of incubation medium the dielectric permeability. This effect is independent of chemical nature of organic solvents which were used with the aim to obtain the corresponding values of D. It is supposed that the cause of activity inhibition of solubilized transporting Ca2+, Mg(2+)-ATPase under the effect of dioxane, acetone, ethanol and inhibition of activity of actomyosine ATPase as affected by dioxane is mainly connected with the increase of electrostatical interaction between opposity charged active centre of ATPase and the product (products) of ATP-hydrolase reaction (Mg ADP-, HPO4(2-)), which is induced by the decrease of incubation medium polarity (the decrease of D value). Stimulating effect of acetone and ethanol on actomyosine ATPase is probably determined by superposition of two components: that connected with direct effect of these solvents on the protein catalyst (interaction with enzyme with the future break of hydrogen and hydrophobic bonds in the protein and its "fluffing") and "electrostatic component" determined by the change of D value of the incubation medium. Possible role of electrostatic interactions between ATPases and reagents as the factor of non-specific control of catalytic activity of these enzymes is discussed.
对一些有机溶剂(2%-10%)影响从平滑肌细胞质膜溶解得到的纯化的转运Ca2+、Mg(2+)-ATP酶以及平滑肌肌动球蛋白ATP酶的作用进行了比较研究。有机溶剂对Ca2+、Mg(2+)-ATP酶初始最大比活性的抑制作用顺序为二氧六环>丙酮>乙醇>二甲基亚砜(DMSO)。与Ca2+、Mg(2+)-ATP酶的情况一样,二氧六环抑制肌动球蛋白ATP酶;丙酮、乙醇和DMSO刺激由收缩蛋白复合物催化的ATP水解反应。已证明,随着孵育介质极性降低,ATP酶活性下降的效应完全由孵育介质的介电常数决定。这种效应与为获得相应介电常数(D值)而使用的有机溶剂的化学性质无关。据推测,在二氧六环、丙酮、乙醇作用下,溶解的转运Ca2+、Mg(2+)-ATP酶活性受到抑制,以及二氧六环影响下肌动球蛋白ATP酶活性受到抑制,其主要原因与孵育介质极性降低(D值降低)引起的ATP酶带相反电荷的活性中心与ATP水解反应产物(Mg ADP-、HPO4(2-))之间静电相互作用增强有关。丙酮和乙醇对肌动球蛋白ATP酶的刺激作用可能由两个因素叠加决定:一个与这些溶剂对蛋白质催化剂的直接作用有关(与酶相互作用,随后蛋白质中的氢键和疏水键断裂并使其“蓬松”),另一个是由孵育介质D值变化决定的“静电因素”。讨论了ATP酶与试剂之间静电相互作用作为这些酶催化活性非特异性控制因素的可能作用。