Duszyk M, French A S, Man S F
Department of Physiology, University of Alberta, Edmonton, Canada.
Biophys J. 1990 Feb;57(2):223-30. doi: 10.1016/S0006-3495(90)82525-X.
The single-channel inside-out patch clamp technique was used to characterize chloride channels in the apical membranes of human airway epithelial cells maintained in primary culture. Patches were obtained from single isolated cells or from cells at the edges of confluent groups. The channel seen most often, in 24% of all patches, had a conductance of approximately 20 pS and had a linear current-voltage relationship in symmetric chloride solutions. The anion selectivity sequence for the channel was NO3- greater than Cl- greater than HCO3-, and it was impermeable to gluconate ions, indicating that the channel diameter lies between 4.7 and 5.5 A. Current through the channel saturated at high chloride concentrations, and the relationship between channel current and chloride concentration could be approximated by the Michaelis-Menten equation. Analysis of the channel's anion permeability and its current vs. concentration relationship indicates that it can be described by the one-ion channel theory, with a relatively weak binding site inside the channel. Histograms of channel open and closed durations were constructed using the log binning technique and could be well fitted by triple exponential distributions, suggesting that the channel has at least three open and three closed states.
采用单通道内面向外式膜片钳技术对原代培养的人气道上皮细胞顶端膜中的氯离子通道进行特性分析。膜片取自单个分离的细胞或汇合细胞群边缘的细胞。在所有膜片中,最常观察到的通道(占24%)电导约为20 pS,在对称氯离子溶液中具有线性电流-电压关系。该通道的阴离子选择性顺序为NO3->Cl->HCO3-,对葡萄糖酸盐离子不通透,表明通道直径在4.7至5.5埃之间。在高氯离子浓度下,通过该通道的电流达到饱和,通道电流与氯离子浓度之间的关系可用米氏方程近似表示。对该通道的阴离子通透性及其电流与浓度关系的分析表明,它可用单离子通道理论来描述,通道内部有一个相对较弱的结合位点。使用对数分箱技术构建了通道开放和关闭持续时间的直方图,这些直方图可以很好地用三重指数分布拟合,表明该通道至少有三个开放状态和三个关闭状态。