高分辨率单通道记录在囊性纤维化突变体功能研究中的应用。
Application of high-resolution single-channel recording to functional studies of cystic fibrosis mutants.
作者信息
Cai Zhiwei, Sohma Yoshiro, Bompadre Silvia G, Sheppard David N, Hwang Tzyh-Chang
机构信息
Department of Physiology and Pharmacology, School of Medical Sciences, University of Bristol, Bristol, UK.
出版信息
Methods Mol Biol. 2011;741:419-41. doi: 10.1007/978-1-61779-117-8_27.
Abstract
The patch-clamp technique is a powerful and versatile method to investigate the cystic fibrosis transmembrane conductance regulator (CFTR) Cl- channel, its malfunction in disease and modulation by small molecules. Here, we discuss how the molecular behaviour of CFTR is investigated using high-resolution single-channel recording and kinetic analyses of channel gating. We review methods used to quantify how cystic fibrosis (CF) mutants perturb the biophysical properties and regulation of CFTR. By explaining the relationship between macroscopic and single-channel currents, we demonstrate how single-channel data provide molecular explanations for changes in CFTR-mediated transepithelial ion transport elicited by CF mutants.
摘要
膜片钳技术是一种强大且通用的方法，用于研究囊性纤维化跨膜传导调节因子（CFTR）氯离子通道、其在疾病中的功能异常以及小分子对其的调节作用。在此，我们讨论如何使用高分辨率单通道记录和通道门控的动力学分析来研究CFTR的分子行为。我们综述了用于量化囊性纤维化（CF）突变体如何扰乱CFTR生物物理特性和调节的方法。通过解释宏观电流和单通道电流之间的关系，我们展示了单通道数据如何为CF突变体引发的CFTR介导的跨上皮离子转运变化提供分子解释。
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