Russell J M, Brodwick M S
Department of Physiology and Biophysics, University of Texas Medical Branch, Galveston 77550.
J Gen Physiol. 1988 Apr;91(4):495-513. doi: 10.1085/jgp.91.4.495.
The intracellular dialysis technique was used to measure unidirectional Cl- fluxes and net acid extrusion by single muscle fibers from the giant barnacle. Decreasing pHi below normal levels of 7.35 stimulated both Cl- efflux and influx. These increases of Cl- fluxes were blocked by disulfonic acid stilbene derivatives such as SITS and DIDS. The SITS-sensitive Cl- efflux was sharply dependent upon pHi, increasing approximately 20-fold as pHi was decreased from 7.35 to 6.7. Under conditions of normal intracellular Mg2+ concentration, the apparent pKa for the activation of Cl- efflux was 7.0. We found that raising [Mg2+]i, but not [Mg2+]o, had a pronounced inhibitory effect on both SITS-sensitive unidirectional Cl- fluxes as well as on SITS-sensitive net acid extrusion. Increasing [Mg2+]i shifted the apparent pKa of Cl- efflux to a more acid value without affecting the maximal flux that could be attained. This relation between pHi and [Mg2+]i on SITS-sensitive Cl- efflux is consistent with a competition between H ions and Mg ions. We conclude that the SITS-inhibitable Cl- fluxes are mediated by the pHi-regulatory transport mechanism and that changes of intracellular Mg2+ levels can modify the activity of the pHi regulator/anion transporter.
采用细胞内透析技术测定来自巨型藤壶的单个肌纤维的单向氯离子通量和净酸排出量。将细胞内pH值降低至正常水平7.35以下会刺激氯离子外流和内流。这些氯离子通量的增加被二磺酸芪衍生物(如SITS和DIDS)阻断。对SITS敏感的氯离子外流强烈依赖于细胞内pH值,当细胞内pH值从7.35降至6.7时,其增加约20倍。在细胞内镁离子浓度正常的条件下,激活氯离子外流的表观pKa为7.0。我们发现,提高细胞内镁离子浓度(而非细胞外镁离子浓度)对SITS敏感的单向氯离子通量以及SITS敏感的净酸排出均有显著抑制作用。增加细胞内镁离子浓度会使氯离子外流的表观pKa向更酸的值移动,而不影响所能达到的最大通量。细胞内pH值和细胞内镁离子浓度对SITS敏感的氯离子外流的这种关系与氢离子和镁离子之间的竞争一致。我们得出结论,SITS可抑制的氯离子通量由细胞内pH值调节转运机制介导,细胞内镁离子水平的变化可改变细胞内pH值调节因子/阴离子转运体的活性。