Department of Chemistry and Biochemistry , The Ohio State University , Columbus , Ohio 43210 , United States.
Center for Information Technology , National Institutes of Health , Bethesda , Maryland 20892 , United States.
J Am Chem Soc. 2018 Oct 17;140(41):13161-13166. doi: 10.1021/jacs.8b06758. Epub 2018 Oct 9.
Application of paramagnetic solid-state NMR to amyloids is demonstrated, using Y145Stop human prion protein modified with nitroxide spin-label or EDTA-Cu tags as a model. By using sample preparation protocols based on seeding with preformed fibrils, we show that paramagnetic protein analogs can be induced into adopting the wild-type amyloid structure. Measurements of residue-specific intramolecular and intermolecular paramagnetic relaxation enhancements enable determination of protein fold within the fibril core and protofilament assembly. These methods are expected to be widely applicable to other amyloids and protein assemblies.
本文展示了顺磁固态 NMR 在淀粉样蛋白中的应用，使用经过氮氧自由基自旋标记或 EDTA-Cu 标记的 Y145Stop 人朊蛋白作为模型。通过使用基于预形成纤维接种的样品制备方案，我们表明可以诱导顺磁蛋白类似物采用野生型淀粉样蛋白结构。残基特异性的分子内和分子间顺磁弛豫增强的测量可以确定纤维核心内的蛋白质折叠和原纤维组装。这些方法有望广泛应用于其他淀粉样蛋白和蛋白质组装体。
