CFTR--ION CONDUCTION AND GATING MECHANISMS

描述：推动这一提议的工作假设是，有 孔内的一个或两个阴离子结合部位，而最好的探针 毛孔本身就是阴离子。实验方法结合 CFTR在单通道宏观非洲爪哇卵母细胞中的表达 正在录音。在某些情况下，可以使用稳定表达cftr的L细胞 (这些细胞将由R.Bridges博士提供)。单声道录制 对于确认电导和摩尔分数效应的变化至关重要 而渗透率可以很容易地从宏观电流确定。 对照将以非CFTR的形式表达卵母细胞和卵母细胞 表达CFTR蛋白，但未受刺激表达电流。 在卵母细胞中发现的内源性IClCa将通过替代 胞外钙离子与钙离子结合。这项建议的重点是阴离子渗透。 独立于CFTR门控问题。这些技术将被用于开发 CFTR的阴离子渗透模型

DESCRIPTION: The working hypothesis driving this proposal is that there are one or two anion binding sites within the pore and that the best probes of the pore are permeant anions themselves. Experimental methods combine expression of CFTR in Xenopus oocytes with single channel and macroscopic recording. In some cases, L-cells which stably express CFTR may be used (these cells will be provided by Dr. R. Bridges). Single channel recording is critical to confirm changes in conductance and mole fraction effects while permeation ratios can be easily determined from macroscopic currents. Controls will be in the form of non-CFTR expressing oocytes and oocytes expressing CFTR protein, but have not been stimulated to express current. The endogenous IClCa found in oocytes will be reduced by replacing extracellular Ca++ with Ba++. This proposal is focused on anion permeation independent of CFTR gating issues. These techniques will be used to develop a model of anion permeation for CFTR.