National High Magnetic Field Laboratory, Tallahassee, Florida 32310, United States.
J Am Chem Soc. 2011 Aug 17;133(32):12370-3. doi: 10.1021/ja204062v. Epub 2011 Jul 26.
The feasibility of using solid-state magic-angle-spinning NMR spectroscopy for in situ structural characterization of the LR11 (sorLA) transmembrane domain (TM) in native Escherichia coli membranes is presented. LR11 interacts with the human amyloid precursor protein (APP), a central player in the pathology of Alzheimer's disease. The background signals from E. coli lipids and membrane proteins had only minor effects on the LR11 TM resonances. Approximately 50% of the LR11 TM residues were assigned by using (13)C PARIS data. These assignments allowed comparisons of the secondary structure of the LR11 TM in native membrane environments and commonly used membrane mimics (e.g., micelles). In situ spectroscopy bypasses several obstacles in the preparation of membrane proteins for structural analysis and offers the opportunity to investigate how membrane heterogeneity, bilayer asymmetry, chemical gradients, and macromolecular crowding affect the protein structure.
本文介绍了使用固态魔角旋转 NMR 光谱法对天然大肠杆菌膜中 LR11（sorLA）跨膜域（TM）进行原位结构表征的可行性。LR11 与人类淀粉样前体蛋白（APP）相互作用，APP 是阿尔茨海默病病理学中的关键参与者。大肠杆菌脂质和膜蛋白的背景信号对 LR11 TM 共振的影响很小。使用（13）C PARIS 数据对大约 50%的 LR11 TM 残基进行了分配。这些分配允许比较在天然膜环境中和常用膜模拟物（例如胶束）中 LR11 TM 的二级结构。原位光谱学避免了在为结构分析制备膜蛋白时的几个障碍，并提供了研究膜异质性、双层不对称性、化学梯度和大分子拥挤如何影响蛋白质结构的机会。
