Division of Pulmonology, Allergy/Immunology, Cystic Fibrosis, and Sleep, Department of Pediatrics, Emory + Children's Center for Cystic Fibrosis and Airways Disease Research, Emory University School of Medicine and Children's Healthcare of Atlanta, Atlanta, GA, USA.
Program in Molecular and Systems Pharmacology, Laney Graduate School, Emory University, Atlanta, GA, USA.
Methods Mol Biol. 2021;2302:49-67. doi: 10.1007/978-1-0716-1394-8_4.
Ion channels play crucial roles in cell physiology, and are a major class of targets for clinically relevant pharmaceuticals. Because they carry ionic current, the function and pharmacology of ion channels can be studied using electrophysiological approaches that range in resolution from the single molecule to many millions of molecules. This chapter describes electrophysiological approaches for the study of one representative ion channel that is defective in a genetic disease, and that is the target of so-called highly effective modulator therapies now used in the clinic: the cystic fibrosis transmembrane conductance regulator (CFTR). Protocols are provided for studying CFTR expressed heterologously, for CFTR expressed in situ in airway epithelial cells, and for purified or partially purified CFTR protein reconstituted into planar lipid bilayers.
离子通道在细胞生理学中起着至关重要的作用,是临床相关药物的主要靶点之一。由于它们携带离子电流,因此可以使用从单个分子到数百万个分子的分辨率的电生理学方法来研究离子通道的功能和药理学。本章描述了用于研究一种遗传性疾病中缺陷的代表性离子通道的电生理学方法,该离子通道也是目前临床应用的所谓高效调节剂治疗的靶标:囊性纤维化跨膜电导调节剂(CFTR)。提供了用于研究异源表达的 CFTR、气道上皮细胞中原位表达的 CFTR 以及重新构成平面脂质双层的纯化或部分纯化 CFTR 蛋白的方案。