NMR Spectroscopy, Bijvoet Centre for Biomolecular Research, Department of Chemistry, Faculty of Science, Utrecht University.
NMR Spectroscopy, Bijvoet Centre for Biomolecular Research, Department of Chemistry, Faculty of Science, Utrecht University;
J Vis Exp. 2021 Mar 3(169). doi: 10.3791/62197.
Membrane proteins are vital for cell function and thus represent important drug targets. Solid-state Nuclear Magnetic Resonance (ssNMR) spectroscopy offers a unique access to probe the structure and dynamics of such proteins in biological membranes of increasing complexity. Here, we present modern solid-state NMR spectroscopy as a tool to study structure and dynamics of proteins in natural lipid membranes and at atomic scale. Such spectroscopic studies profit from the use of high-sensitivity ssNMR methods, i.e., proton-(H)-detected ssNMR and DNP (Dynamic Nuclear Polarization) supported ssNMR. Using bacterial outer membrane beta-barrel protein BamA and the ion channel KcsA, we present methods to prepare isotope-labeled membrane proteins and to derive structural and motional information by ssNMR.
膜蛋白对于细胞功能至关重要，因此是重要的药物靶点。固态核磁共振（ssNMR）光谱学为研究越来越复杂的生物膜中此类蛋白质的结构和动力学提供了一种独特的方法。在这里，我们将现代固态 NMR 光谱学作为一种工具来研究天然脂质膜中和原子尺度上的蛋白质结构和动力学。这些光谱学研究受益于使用高灵敏度的 ssNMR 方法，即质子（H）检测的 ssNMR 和 DNP（动态核极化）支持的 ssNMR。使用细菌外膜β桶状蛋白 BamA 和离子通道 KcsA，我们介绍了制备同位素标记的膜蛋白的方法，并通过 ssNMR 获得结构和动态信息。
