DNP-Enhanced MAS NMR: A Tool to Snapshot Conformational Ensembles of α-Synuclein in Different States.
DNP-Enhanced MAS NMR: A Tool to Snapshot Conformational Ensembles of α-Synuclein in Different States.
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DOI：
10.1016/j.bpj.2018.02.011
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发表时间：
2018-04-10
影响因子：
3.4
通讯作者：
Heise H
中科院分区：
文献类型：
--
作者：
Uluca B;Viennet T;Petrović D;Shaykhalishahi H;Weirich F;Gönülalan A;Strodel B;Etzkorn M;Hoyer W;Heise H
本质上无序的蛋白质动态采样广泛的构象空间，因此不采用稳定且明确的三维构象。结构异质性与其在生理过程中的正常功能有关。了解构象集合对于完全理解此类蛋白质至关重要。我们在这里提出一种方法，利用冷冻溶液中稀疏同位素标记蛋白质的动态核极化增强固态核磁共振波谱，通过利用不均匀加宽的线形状来拍摄完整结构整体的快照。我们研究了本质上无序的蛋白质 α-突触核蛋白 (α-syn)，它在帕金森病的病因学中发挥着关键作用，处于三种不同的生理相关状态。对于冷冻溶液中的游离单体，我们可以看到所谓的“无规卷曲构象”由α螺旋和β片状构象组成，并且相邻氨基酸的二级化学位移往往是相关的，表明二级结构元件的频繁形成。基于这些结果，我们可以估计纤维状 α-syn 以及以不同蛋白质与脂质比率结合到膜上的 α-syn 中无序区域的数量。因此，我们的方法提供了有关在不同功能状态下采样瞬态二级结构的倾向的定量信息。分子动力学模拟使结果合理化。
Intrinsically disordered proteins dynamically sample a wide conformational space and therefore do not adopt a stable and defined three-dimensional conformation. The structural heterogeneity is related to their proper functioning in physiological processes. Knowledge of the conformational ensemble is crucial for a complete comprehension of this kind of proteins. We here present an approach that utilizes dynamic nuclear polarization-enhanced solid-state NMR spectroscopy of sparsely isotope-labeled proteins in frozen solution to take snapshots of the complete structural ensembles by exploiting the inhomogeneously broadened line-shapes. We investigated the intrinsically disordered protein α-synuclein (α-syn), which plays a key role in the etiology of Parkinson’s disease, in three different physiologically relevant states. For the free monomer in frozen solution we could see that the so-called “random coil conformation” consists of α-helical and β-sheet-like conformations, and that secondary chemical shifts of neighboring amino acids tend to be correlated, indicative of frequent formation of secondary structure elements. Based on these results, we could estimate the number of disordered regions in fibrillar α-syn as well as in α-syn bound to membranes in different protein-to-lipid ratios. Our approach thus provides quantitative information on the propensity to sample transient secondary structures in different functional states. Molecular dynamics simulations rationalize the results.
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DOI：
10.1073/pnas.0407146102
发表时间：
2005-02-01
影响因子：
11.1
作者：
Bertoncini, CW;Jung, YS;Zweckstetter, M
通讯作者：
Zweckstetter, M
影响因子：
5.5
作者：
Beauchamp, Kyle A.;Lin, Yu-Shan;Das, Rhiju;Pande, Vijay S.
通讯作者：
Pande, Vijay S.
影响因子：
1.7
作者：
Bussi, Giovanni
通讯作者：
Bussi, Giovanni
影响因子：
5.5
作者：
Best, Robert B.;Zhu, Xiao;Shim, Jihyun;Lopes, Pedro E. M.;Mittal, Jeetain;Feig, Michael;MacKerell, Alexander D., Jr.
通讯作者：
MacKerell, Alexander D., Jr.
影响因子：
4.4
作者：
Cavalli, Andrea;Camilloni, Carlo;Vendruscolo, Michele
通讯作者：
Vendruscolo, Michele