University of Texas Health Science Center at San Antonio, San Antonio, TX, USA.
Methods Mol Biol. 2024;2796:271-289. doi: 10.1007/978-1-0716-3818-7_16.
Ion channels are membrane proteins that may also have intracellular and extracellular domains that interact with other ligands. In many cases, these interaction sites are highly mobile and may undergo changes in the configuration upon binding with regulatory signaling molecules. Isothermal titration calorimetry (ITC) is a powerful technique to quantify protein-ligand interactions of purified samples in solution. This chapter describes a fragment-based analysis method using ITC to quantify the interactions between a domain of the voltage-gated Kv7 channel and the calcium-regulated protein calmodulin. This example can be used to quantify the interactions between specific domains of other ion channels and their regulatory signaling proteins.
离子通道是一种膜蛋白，它可能还有细胞内和细胞外的结构域与其他配体相互作用。在许多情况下，这些相互作用位点具有高度的移动性，并且在与调节信号分子结合时可能会发生构象变化。等温滴定量热法（ITC）是一种强大的技术，可用于定量分析溶液中纯化样品中蛋白质-配体的相互作用。本章描述了一种使用 ITC 进行基于片段分析的方法，用于量化电压门控 Kv7 通道的一个结构域与钙调节蛋白钙调蛋白之间的相互作用。这个例子可以用于量化其他离子通道的特定结构域与其调节信号蛋白之间的相互作用。
