NMR Spectroscopy, Bijvoet Center for Biomolecular Research, Utrecht University, 3584 CH Utrecht, the Netherlands; Oxford Particle Imaging Centre, Division of Structural Biology, University of Oxford, The Wellcome Trust Centre for Human Genetics, Roosevelt Drive, Oxford OX3 7BN, UK.
NMR Spectroscopy, Bijvoet Center for Biomolecular Research, Utrecht University, 3584 CH Utrecht, the Netherlands.
Structure. 2018 Jan 2;26(1):161-170.e3. doi: 10.1016/j.str.2017.11.011. Epub 2017 Dec 14.
Membrane proteins remain challenging targets for structural biology, despite much effort, as their native environment is heterogeneous and complex. Most methods rely on detergents to extract membrane proteins from their native environment, but this removal can significantly alter the structure and function of these proteins. Here, we overcome these challenges with a hybrid method to study membrane proteins in their native membranes, combining high-resolution solid-state nuclear magnetic resonance spectroscopy and electron cryotomography using the same sample. Our method allows the structure and function of membrane proteins to be studied in their native environments, across different spatial and temporal resolutions, and the combination is more powerful than each technique individually. We use the method to demonstrate that the bacterial membrane protein YidC adopts a different conformation in native membranes and that substrate binding to YidC in these native membranes differs from purified and reconstituted systems.
尽管付出了很多努力，膜蛋白仍然是结构生物学的一个具有挑战性的目标，因为它们的天然环境是异质和复杂的。大多数方法依赖于去污剂从天然环境中提取膜蛋白，但这种去除会显著改变这些蛋白质的结构和功能。在这里，我们通过一种混合方法克服了这些挑战，该方法用于在天然膜中研究膜蛋白，将高分辨率固态核磁共振波谱学和电子晶体断层扫描技术结合使用相同的样品。我们的方法允许在不同的空间和时间分辨率下研究膜蛋白的结构和功能，并且这种组合比每个单独的技术都更强大。我们使用该方法证明细菌膜蛋白 YidC 在天然膜中采用不同的构象，并且在这些天然膜中底物与 YidC 的结合与纯化和重组系统不同。
