19F NMR as a versatile tool to study membrane protein structure and dynamics
19F NMR as a versatile tool to study membrane protein structure and dynamics
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DOI：
10.1515/hsz-2018-0473
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发表时间：
2019-09-01
影响因子：
3.7
通讯作者：
Hellmich, Ute A.
中科院分区：
文献类型：
--
作者：
Rose-Sperling, Dania;Mai Anh Tran;Hellmich, Ute A.
为了阐明膜蛋白的结构和动力学，已经发展了非常先进的生物物理方法，这些方法通常需要大量的资源，无论是样品制备还是实验分析。对于非常复杂的系统，如膜转运体、离子通道或G蛋白偶联受体(GPCRs)，在选定的位置加入单个报告可以显著简化观察和测量/分析要求。在这里，我们列举了使用F-19核磁共振光谱作为研究(膜)蛋白质结构、动力学和配基相互作用的强大而相对简单的工具的例子。我们总结了将F-19标记结合到蛋白质中的方法，并讨论了从相对简单的核磁共振谱中可以容易地从膜蛋白质中获得的信息类型，重点是GPCRs作为该技术研究最广泛的膜蛋白家族。在未来，这些方法可能还会对许多经历复杂功能动力学和/或包含非结构化区域的蛋白质产生特别的兴趣，因此不适用于X射线结晶学或冷冻电子显微镜(CryoEM)的研究。
To elucidate the structures and dynamics of membrane proteins, highly advanced biophysical methods have been developed that often require significant resources, both for sample preparation and experimental analyses. For very complex systems, such as membrane transporters, ion channels or G-protein coupled receptors (GPCRs), the incorporation of a single reporter at a select site can significantly simplify the observables and the measurement/analysis requirements. Here we present examples using F-19 nuclear magnetic resonance (NMR) spectroscopy as a powerful, yet relatively straightforward tool to study (membrane) protein structure, dynamics and ligand interactions. We summarize methods to incorporate F-19 labels into proteins and discuss the type of information that can be readily obtained for membrane proteins already from relatively simple NMR spectra with a focus on GPCRs as the membrane protein family most extensively studied by this technique. In the future, these approaches may be of particular interest also for many proteins that undergo complex functional dynamics and/or contain unstructured regions and thus are not amenable to X-ray crystallography or cryo electron microscopy (cryoEM) studies.