Spin Labeling and Characterization of Tau Fibrils Using Electron Paramagnetic Resonance (EPR).
Spin Labeling and Characterization of Tau Fibrils Using Electron Paramagnetic Resonance (EPR).
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DOI：
10.1007/978-1-4939-2978-8_12
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发表时间：
2016
影响因子：
--
通讯作者：
Virginia Meyer;M. Margittai
中科院分区：
文献类型：
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作者：
Virginia Meyer;M. Margittai
Tau原纤维的模板辅助增殖对于阿尔茨海默病中Tau病理学的传播是必不可少的。在这个过程中，原纤维的小种子将Tau单体募集到它们的末端。纤维的物理性质在其生长过程中起着重要作用。在这里，我们描述了两种不同的电子顺磁共振（EPR）技术，提供了至关重要的见解的结构的Tau纤维。这两种技术都依赖于将一个或两个自旋标记定点引入蛋白质单体中。连续波（CW）EPR提供了关于纤维核心中包含哪些氨基酸残基以及它们如何沿着长纤维轴堆叠的信息。双电子-电子共振（DEER）确定了单个蛋白质内两个自旋标记之间的距离，因此提供了对它们在原纤维横截面中的空间排列的见解。由于DEER（~2-5 nm）可接近的距离较长，因此可以区分不同原纤维构象的种群。
Template-assisted propagation of Tau fibrils is essential for the spreading of Tau pathology in Alzheimer’s disease. In this process, small seeds of fibrils recruit Tau monomers onto their ends. The physical properties of the fibrils play an important role in their propagation. Here, we describe two different electron paramagnetic resonance (EPR) techniques that have provided crucial insights into the structure of Tau fibrils. Both techniques rely on the site-directed introduction of one or two spin labels into the protein monomer. Continuous-wave (CW) EPR provides information on which amino acid residues are contained in the fibril core and how they are stacked along the long fibril axis. Double electron–electron resonance (DEER) determines distances between two spin labels within a single protein and hence provides insights into their spatial arrangement in the fibril cross section. Because of the long distance range accessible to DEER (~2–5 nm) populations of distinct fibril conformers can be differentiated.