Béliveau R, Strévey J
Départment de Chimie, Université du Québec à Montréal, Canada.
Am J Physiol. 1988 Mar;254(3 Pt 2):F329-36. doi: 10.1152/ajprenal.1988.254.3.F329.
Phosphate transport was studied in brush-border membrane vesicles purified from rat kidney cortex. Influx and efflux were strongly dependent on the presence of cis sodium; the rate of efflux, calculated by linear regression performed on the first time points, was much lower than the rate of influx (0.044 vs. 0.198 pmol.microgram protein-1.s-1). Trans phosphate had a stimulatory effect on phosphate influx (145% stimulation at 10 mM phosphate trans, with 0.2 mM phosphate cis). Trans phosphate was, however, inhibitory for phosphate efflux (89% inhibition at 10 mM phosphate trans). Trans effects of sodium were also studied. With 200 mM trans sodium, we observed 73% inhibition of phosphate influx and 60% inhibition of phosphate efflux. Studies involving sodium and phosphate present at the same time as trans substrates showed that the trans inhibition of phosphate influx by sodium could be completely reversed by trans phosphate. Trans inhibition of phosphate efflux by phosphate was not additive to the inhibition caused by sodium. Addition of trans phosphate had a stimulatory effect on sodium-independent influx, indicating that the binary complex (C-P) could translocate in efflux. These results indicate that the renal phosphate carrier presents a random binding scheme for the intra- and extravesicular sides of the membrane.
对从大鼠肾皮质纯化的刷状缘膜囊泡中的磷酸盐转运进行了研究。内流和外流强烈依赖于顺式钠的存在;通过对最初时间点进行线性回归计算得出的外流速率远低于内流速率(分别为0.044和0.198 pmol·μg蛋白⁻¹·s⁻¹)。反式磷酸盐对磷酸盐内流有刺激作用(在10 mM反式磷酸盐、0.2 mM顺式磷酸盐存在时刺激145%)。然而,反式磷酸盐对磷酸盐外流有抑制作用(在10 mM反式磷酸盐时抑制89%)。还研究了钠的反式效应。在200 mM反式钠存在时,我们观察到磷酸盐内流抑制73%,磷酸盐外流抑制60%。涉及钠和磷酸盐同时作为反式底物存在的研究表明,钠对磷酸盐内流的反式抑制可被反式磷酸盐完全逆转。磷酸盐对磷酸盐外流的反式抑制与钠引起的抑制无相加作用。添加反式磷酸盐对不依赖钠的内流有刺激作用,表明二元复合物(C-P)可在外流中转运。这些结果表明,肾磷酸盐载体对膜的囊泡内和囊泡外两侧呈现随机结合模式。