Zifarelli Giovanni, Soliani Paolo, Pusch Michael
Istituto di Biofisica, Consiglio Nazionale delle Ricerche, Genoa, Italy.
Biophys J. 2008 Jan 1;94(1):53-62. doi: 10.1529/biophysj.107.111310. Epub 2007 Sep 7.
H(+) ions are a substrate of many active and passive membrane transporters in all cells. Absolute proton fluxes are often quantified using intracellular pH sensitive microelectrodes or pH sensitive dyes. These measurements, however, rely on a priori estimates of the intracellular buffer capacity and on the assumption of diffusive equilibrium inside the cell. Here, assuming local equilibrium of protons with a single mobile buffer, we model the diffusion of H(+) in the extracellular medium around an H(+) pumping cell to estimate the expected pH changes as a function of time, distance from the cell, extracellular buffer capacity, and the absolute proton flux across the membrane. In particular, using accurate numerical simulation, we gauge the range of validity of an explicit, analytical solution of the linearized, nonstationary diffusion equation. Our results provide a framework to quantify the absolute membrane proton flux, if spatiotemporal information about the extracellular pH change is available, e.g., using imaging of pH dependent fluorescent dyes.
氢离子是所有细胞中许多主动和被动膜转运蛋白的底物。绝对质子通量通常使用细胞内pH敏感微电极或pH敏感染料进行量化。然而,这些测量依赖于细胞内缓冲能力的先验估计以及细胞内扩散平衡的假设。在这里,假设质子与单一移动缓冲剂处于局部平衡,我们对氢离子在氢离子泵浦细胞周围的细胞外介质中的扩散进行建模,以估计预期的pH变化作为时间、与细胞的距离、细胞外缓冲能力以及跨膜绝对质子通量的函数。特别是,通过精确的数值模拟,我们评估了线性化非平稳扩散方程显式解析解的有效性范围。如果有关于细胞外pH变化的时空信息,例如使用pH依赖性荧光染料成像,我们的结果提供了一个量化绝对膜质子通量的框架。