Orsenigo M N, Tosco M, Zoppi S, Faelli A
Dipartimento di Fisiologia e Biochimica Generali, Università di Milano, Italy.
Biochim Biophys Acta. 1990 Jul 9;1026(1):64-8. doi: 10.1016/0005-2736(90)90333-j.
Na uptake studies were performed in order to examine the activity of a Na/H exchanger in basolateral membrane vesicles isolated from rat jejunum. Experiments were carried out under voltage-clamped conditions in order to avoid electrodiffusional ionic movements. 1 mM Na uptake was found to be enhanced by an outward proton gradient and its initial rate was further increased by the presence of monensin or nigericin. The pH gradient-driven Na uptake was inhibited by 2 mM amiloride and unaffected by 0.1 mM 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid. The initial rate of the proton gradient-induced Na uptake was saturable with respect to external Na, with a Km of 13.6 +/- 1.4 mM and a Vmax of 35.4 +/- 2.2 nmol/mg protein per min. Li competed with Na for the exchange process, whereas K, Rb, Cs, tetramethylammonium had no effect. We conclude that rat jejunal basolateral membrane contains a Na/H exchanger whose properties are similar to those of the antiporter identified in the brush-border membrane.
为了检测从大鼠空肠分离的基底外侧膜囊泡中钠/氢交换体的活性,进行了钠摄取研究。实验在电压钳制条件下进行,以避免离子的电扩散运动。发现1 mM的钠摄取因外向质子梯度而增强,其初始速率因莫能菌素或尼日利亚菌素的存在而进一步增加。pH梯度驱动的钠摄取受到2 mM氨氯吡咪的抑制,不受0.1 mM 4,4'-二异硫氰酸芪-2,2'-二磺酸的影响。质子梯度诱导的钠摄取的初始速率相对于外部钠是可饱和的,Km为13.6±1.4 mM,Vmax为35.4±2.2 nmol/mg蛋白质每分钟。锂在交换过程中与钠竞争,而钾、铷、铯、四甲基铵则没有影响。我们得出结论,大鼠空肠基底外侧膜含有一种钠/氢交换体,其特性与在刷状缘膜中鉴定的反向转运体相似。