Bucuvalas J C, Goodrich A L, Suchy F J
Am J Physiol. 1987 Sep;253(3 Pt 1):G351-8. doi: 10.1152/ajpgi.1987.253.3.G351.
Highly purified rat basolateral liver plasma membrane vesicles were used to examine the mechanism and the driving forces for hepatic uptake of the beta-amino acid, taurine. An inwardly directed 100 mM NaCl gradient stimulated the initial rate of taurine uptake and energized a transient twofold accumulation of taurine above equilibrium ("overshoot"). In contrast, uptake was slower and no overshoot was detected in the presence of a KCl gradient. A negative intravesicular electrical potential generated by the presence of permeant anions or an outwardly directed K+ gradient with valinomycin increased Na+-stimulated taurine uptake. External Cl- stimulated Na+-dependent taurine uptake independent of effects on the transmembrane electrical potential difference. Na+-dependent taurine uptake showed a sigmoidal dependence on extravesicular Na+ concentration, suggesting multiple Na+ ions are involved in the translocation of each taurine molecule. Na+-dependent taurine uptake demonstrated Michaelis-Menten kinetics with a maximum velocity of 0.537 nmol.mg protein-1.min-1 and an apparent Km of 174 microM. [3H]taurine uptake was inhibited by the presence of excess unlabeled taurine, beta-alanine, or hypotaurine but not by L-glutamine or L-alanine. In summary, using basolateral liver plasma membrane vesicles, we have shown that hepatic uptake of taurine occurs by a carrier-mediated, secondary active transport process specific for beta-amino acids. Uptake is electrogenic, stimulated by external Cl-, and requires multiple Na+ ions for the translocation of each taurine molecule.
使用高度纯化的大鼠肝基底外侧质膜囊泡来研究β-氨基酸牛磺酸的肝摄取机制和驱动力。内向的100 mM NaCl梯度刺激了牛磺酸摄取的初始速率,并使牛磺酸在平衡状态以上出现了两倍的瞬时积累(“过冲”)。相比之下,在存在KCl梯度的情况下,摄取较慢且未检测到过冲现象。由渗透性阴离子的存在或与缬氨霉素形成的外向K +梯度产生的囊泡内负电势增加了Na +刺激的牛磺酸摄取。外部Cl-刺激了Na +依赖性牛磺酸摄取,而与对跨膜电势差的影响无关。Na +依赖性牛磺酸摄取对细胞外Na +浓度呈S形依赖性,表明每个牛磺酸分子的转运涉及多个Na +离子。Na +依赖性牛磺酸摄取表现出米氏动力学,最大速度为0.537 nmol·mg蛋白-1·min-1,表观Km为174 μM。过量未标记的牛磺酸、β-丙氨酸或亚牛磺酸的存在会抑制[3H]牛磺酸摄取,但L-谷氨酰胺或L-丙氨酸则不会。总之,使用肝基底外侧质膜囊泡,我们表明牛磺酸的肝摄取是通过载体介导的、对β-氨基酸特异的继发性主动转运过程发生的。摄取是电生的,受外部Cl-刺激,并且每个牛磺酸分子的转运需要多个Na +离子。