Reichenwallner Jörg, Liu Boyuan, Balo Aidin R, Ou Wei-Lin, Ernst Oliver P
Department of Biochemistry, University of Toronto, Toronto, ON, M5S 1A8, Canada.
Department of Biochemistry, University of Toronto, Toronto, ON, M5S 1A8, Canada.
Curr Opin Struct Biol. 2021 Aug;69:177-186. doi: 10.1016/j.sbi.2021.06.003. Epub 2021 Jul 23.
Membrane proteins, including ion channels, transporters and G-protein-coupled receptors (GPCRs), play a significant role in various physiological processes. Many of these proteins are difficult to express in large quantities, imposing crucial experimental restrictions. Nevertheless, there is now a wide variety of studies available utilizing electron paramagnetic resonance (EPR) spectroscopic techniques that expand experimental accessibility by using relatively small quantities of protein. Here, we give an overview starting from basic strategies in EPR on membrane proteins with a focus on GPCRs, while emphasizing several applications from recent years. We highlight how the arsenal of EPR-based techniques may provide significant further contributions to understanding the complex molecular machinery and energetic phenomena responsible for seamless workflow in essential biological processes.
膜蛋白,包括离子通道、转运蛋白和G蛋白偶联受体(GPCRs),在各种生理过程中发挥着重要作用。其中许多蛋白质难以大量表达,这对实验造成了关键限制。然而,现在有各种各样利用电子顺磁共振(EPR)光谱技术的研究,这些技术通过使用相对少量的蛋白质来扩大实验的可及性。在这里,我们从EPR研究膜蛋白的基本策略开始进行概述,重点是GPCRs,同时强调近年来的一些应用。我们强调基于EPR的技术如何为理解负责基本生物过程中无缝工作流程的复杂分子机制和能量现象做出重大进一步贡献。