Meznikov S A, Meznikova O V
Izv Akad Nauk Ser Biol. 2005 Jan-Feb(1):18-22.
We studied the effect of furosemide on GABAA-induced 36Cl transport and GABAA-induced Cl- -ATPase activity in synaptic membranes of fish brain. At physiological pH of 7.4, GABA (0.1-100 microM) stimulated 36Cl transport in synaptoneurosomes and Cl- -ATPase activity in synaptic membranes. Furosemide (0.1-0.5 mM) removed the activating effect of the mediator on chlorine transport and enzyme activity (I50 equaled 0.16 and 0.12 mM, respectively). In the absence of the mediator, picrotoxin (50 microM) activated the basal 36Cl influx in synaptoneurosomes and the basal Mg2+ -ATPase activity of synaptic membranes. Furosemide (1 mM) removed the activating effect of picrotoxin on both biochemical processes. The obtained data demonstrated similar sensitivities of GABAA-induced transport of 36Cl in synaptoneurosomes and of GABAA-induced Cl- -ATPase activity in synaptic membranes to furosemide and indicated the involvement of the ATPase in GABAA-induced processes. The soluble ATPase, recovered by sodium deoxycholate solubilization of the membranes, remained sensitive to GABAA-ergic ligands, which suggested proximity of their binding sites with ATP hydrolysis sites of protein molecule and their structural contingency.
我们研究了呋塞米对鱼脑突触膜中GABAA诱导的36Cl转运和GABAA诱导的Cl--ATP酶活性的影响。在生理pH值7.4时,GABA(0.1 - 100微摩尔)刺激突触神经小体中的36Cl转运以及突触膜中的Cl--ATP酶活性。呋塞米(0.1 - 0.5毫摩尔)消除了介质对氯转运和酶活性的激活作用(I50分别等于0.16和0.12毫摩尔)。在没有介质的情况下,印防己毒素(50微摩尔)激活突触神经小体中的基础36Cl内流以及突触膜的基础Mg2+-ATP酶活性。呋塞米(1毫摩尔)消除了印防己毒素对这两个生化过程的激活作用。所获得的数据表明,突触神经小体中GABAA诱导的36Cl转运以及突触膜中GABAA诱导的Cl--ATP酶活性对呋塞米具有相似的敏感性,并表明ATP酶参与了GABAA诱导的过程。通过用脱氧胆酸钠溶解膜回收的可溶性ATP酶对GABAA能配体仍保持敏感,这表明它们的结合位点与蛋白质分子的ATP水解位点接近且在结构上相关。