Szollosi Andras
Semmelweis University, Department of Biochemistry, Budapest, Hungary.
HUN-REN-SE Ion Channel Research Group, Budapest, Hungary.
Methods Mol Biol. 2025;2908:141-161. doi: 10.1007/978-1-0716-4434-8_10.
Ion channels form transmembrane protein pores allowing ion fluxes across biological membranes. Their function is indispensable for normal homeostasis; therefore, channelopathies lead to a large variety of diseases affecting almost all tissues and organs. In the last decades, the structure of hundreds of ion channels has been solved. In the majority of the solved structures the pore is closed, and for most of the protein preparations used in these studies, currently little data is available to support that the purified protein forms a functional ion channel capable of gating. Planar lipid bilayer technique is a powerful tool to address this issue. Purified ion channel protein is incorporated into a membrane bilayer separating two compartments in an experimental chamber. Channel currents are recorded between the two sides. Such setup allows characterization of gating and permeation properties of ion channels unbiased by auxiliary components present in native membranes. The planar lipid bilayer technique is thus discussed herein together with the robust protein expressing BacMam system.
离子通道形成跨膜蛋白孔道，允许离子通量穿过生物膜。它们的功能对于正常的体内平衡不可或缺；因此，通道病会导致影响几乎所有组织和器官的多种疾病。在过去几十年中，数百种离子通道的结构已被解析。在大多数已解析的结构中，孔道是关闭的，并且对于这些研究中使用的大多数蛋白质制剂，目前几乎没有数据支持纯化的蛋白质形成能够门控的功能性离子通道。平面脂质双层技术是解决这个问题的有力工具。纯化的离子通道蛋白被整合到分隔实验腔室中两个隔室的膜双层中。在两侧记录通道电流。这样的设置允许表征离子通道的门控和通透特性，而不受天然膜中存在的辅助成分的影响。因此，本文将讨论平面脂质双层技术以及强大的蛋白质表达BacMam系统。
