Jacobsen C, Kragh-Hansen U, Sheikh M I
Biochem J. 1986 Oct 15;239(2):411-6. doi: 10.1042/bj2390411.
Na+-H+-exchanger activity of pars convoluta and pars recta luminal-membrane vesicles prepared from the proximal tubule of acidotic and control rabbits were assayed by a rapid-filtration technique and an Acridine Orange method. Both experimental approaches revealed the existence of an antiporter, sensitive to metabolic acidosis, in pars convoluta membrane vesicles. Kinetic data, obtained with the pH-sensitive dye, showed that the Km for Na+ transport was unchanged by acidosis, whereas Vmax. for exchanger activity was increased, on an average, by 44%. The fluorescence method, in contrast with the rapid-filtration technique, was able to detect exchanger activity in pars recta membrane vesicles. The Km value for the antiporter located in pars recta is comparable with that calculated for pars convoluta membrane vesicles. By contrast, the Vmax. of this exchanger is only about 25% of that found for pars convoluta. Furthermore, metabolic acidosis apparently does not increase Na+-H+-exchanger activity of pars recta luminal-membrane vesicles.
采用快速过滤技术和吖啶橙法,对酸中毒兔和对照兔近端小管制备的曲部和直部管腔膜囊泡的Na⁺-H⁺交换活性进行了测定。两种实验方法均显示,曲部膜囊泡中存在一种对代谢性酸中毒敏感的反向转运体。用pH敏感染料获得的动力学数据表明,酸中毒时Na⁺转运的Km值未发生变化,而交换体活性的Vmax平均增加了44%。与快速过滤技术相比,荧光法能够检测直部膜囊泡中的交换体活性。位于直部的反向转运体的Km值与曲部膜囊泡计算得出的Km值相当。相比之下,该交换体的Vmax仅约为曲部的25%。此外,代谢性酸中毒显然不会增加直部管腔膜囊泡的Na⁺-H⁺交换活性。