Jonas A J, Jobe H
Department of Pediatrics, Harbor-UCLA Medical Center, Torrance 90502.
J Biol Chem. 1990 Oct 15;265(29):17545-9.
Sulfate transport was examined using membrane vesicles (pH 7.0 inside) prepared from rat liver lysosomes. Sulfate uptake was dependent upon external pH with increased uptake at lower buffer pH. The Km for uptake was 160 microM at pH 5.0 while at pH 7.0, a lower affinity system with a Km of 1.4 mM was present. The protonophore carbonyl cyanide m-chlorophenylhydrazone increased uptake at pH 5.0 while valinomycin/KCl had no effect. In contrast, at pH 7.0, valinomycin-induced changes in membrane potential stimulated uptake. Countertransport of sulfate at pH 7.0 was inhibited by 4,4'-diisothiocyano-2,2'-disulfonic acid stilbene, N-(4-azido-2-nitrophenyl)-2-aminoethanesulfonic acid, and a variety of anions: SO4(2-) greater than MoO4(2-) greater than Cl- greater than HPO4- greater than HCO3-. Trans-stimulation of sulfate uptake at pH 7.0 was observed with MoO4(2-) and, to a lesser extent, with S2O3(2-) while Cl-, HPO4-, and HCO3- had little effect. However, chloride loading of vesicles resulted in marked stimulation of sulfate uptake at pH 5.0. It appears that sulfate and protons exit lysosomes in exchange for chloride by a specific, pH-regulated anion transport system.
利用从大鼠肝脏溶酶体制备的膜泡(内部pH值为7.0)研究了硫酸盐转运。硫酸盐摄取依赖于外部pH值,在较低缓冲液pH值下摄取增加。在pH 5.0时摄取的Km为160微摩尔,而在pH 7.0时,存在一个亲和力较低的系统,Km为1.4毫摩尔。质子载体羰基氰化物间氯苯腙在pH 5.0时增加摄取,而缬氨霉素/KCl没有影响。相反,在pH 7.0时,缬氨霉素诱导的膜电位变化刺激了摄取。在pH 7.0时,硫酸盐的反向转运受到4,4'-二异硫氰酸-2,2'-二磺酸芪、N-(4-叠氮基-2-硝基苯基)-2-氨基乙烷磺酸以及多种阴离子的抑制:SO4(2-)>MoO4(2-)>Cl->HPO4->HCO3-。在pH 7.0时,观察到用MoO4(2-)以及在较小程度上用S2O3(2-)对硫酸盐摄取有反刺激作用,而Cl-、HPO4-和HCO3-几乎没有影响。然而,膜泡加载氯化物导致在pH 5.0时显著刺激硫酸盐摄取。似乎硫酸盐和质子通过一个特定的、pH调节的阴离子转运系统与氯化物交换而从溶酶体中排出。