Chernysh I H
O.V. Palladin Institute of Biochemistry of NAS of Ukraine, Kyiv.
Ukr Biokhim Zh (1999). 1999 Sep-Oct;71(5):123-6.
The eosin Y inhibitory effect on the activity of smooth muscle plasma membrane Ca(2+)-transporting ATPase was studied: effect of this inhibitor on the maximal initial rate of ATP-hydrolase reaction, catalyzed by Ca2+, Mg(2+)-ATPase, on the affinity of enzyme for the reaction reagents (Ca2+, Mg2+, ATP). Dependence of eosin Y inhibitory effect on some physicochemical factors of incubation medium was studied too. It was determined that eosin Y inhibited reversibly and with high specificity purified Ca2+, Mg(2+)-ATPase solubilized from myometrial cell plasma membrane (Ki--0.8 microM), decreased the turnover rate of this enzyme determined both by Mg2+, ATP and Ca2+. This inhibitor had no effect on the enzyme affinity for Ca2+, increased affinity for Mg2+ and decreased affinity for ATP. It was determined that inhibition of Ca2+, Mg(2+)-ATPase by eosin Y depended on pH and dielectric permeability of the incubation medium: increasing of pH from 6.5 to 8.0 reduced the apparent Ki, decreasing of dielectric permeability from 74.07 to 71.19 increased the apparent Ki.
研究了伊红Y对平滑肌质膜Ca(2+)-转运ATP酶活性的抑制作用:该抑制剂对由Ca2+、Mg(2+)-ATP酶催化的ATP水解酶反应的最大初始速率、酶对反应试剂(Ca2+、Mg2+、ATP)的亲和力的影响。还研究了伊红Y抑制作用对孵育介质某些物理化学因素的依赖性。确定伊红Y可逆且高度特异性地抑制从子宫肌层细胞质膜溶解的纯化Ca2+、Mg(2+)-ATP酶(Ki--0.8 microM),降低了由Mg2+、ATP和Ca2+测定的该酶的周转速率。该抑制剂对酶对Ca2+的亲和力没有影响,增加了对Mg2+的亲和力并降低了对ATP的亲和力。确定伊红Y对Ca2+、Mg(2+)-ATP酶的抑制作用取决于孵育介质的pH值和介电常数:pH值从6.5增加到8.0降低了表观Ki,介电常数从74.07降低到71.19增加了表观Ki。