Conformational fingerprinting of tau variants and strains by Raman spectroscopy.
Conformational fingerprinting of tau variants and strains by Raman spectroscopy.
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DOI：
10.1039/d1ra00870f
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发表时间：
2021
期刊：
影响因子：
3.9
通讯作者：
Mudher A
中科院分区：
文献类型：
--
作者：
Devitt G;Crisford A;Rice W;Weismiller HA;Fan Z;Commins C;Hyman BT;Margittai M;Mahajan S;Mudher A
tau蛋白病是一组异常折叠的tau蛋白沉积伴随神经变性的疾病。发展蛋白质构象的病理变化的检测和分类方法对于理解影响Tau蛋白病聚集体结构多态性的因素是期望的。我们以前已经证明了拉曼光谱的实用性，用于表征和区分不同的蛋白质聚集体，包括tau，基于其独特的构象签名。在此基础上，在本研究中，我们评估了拉曼光谱的效用，用于表征和区分不同的构象的相同的蛋白质，在这种情况下的tau是独特的tau菌株在体外产生的。我们现在研究聚集环境、辅因子、翻译后修饰和一级序列对tau纤维的拉曼指纹的影响。使用定量构象指纹和多元统计分析，我们发现，在不同的缓冲液条件下的tau蛋白的聚集导致形成不同的原纤维菌株。鉴定了使用肝素或RNA辅因子产生的tau原纤维以及磷酸化tau的独特光谱标志物。我们还确定了tau单体的一级序列影响所得tau原纤维的构象特征，包括2N4R、0N3R、K18和P301S tau变体。这些结果突出了tau原纤维的构象多态性，这反映在广泛的相关神经系统疾病中。此外，在这项研究中提出的分析提供了一个基准的拉曼光谱表征的tau菌株，这可能会揭示聚集环境，辅因子和翻译后修饰如何影响tau构象在体内的未来研究。我们报告说，聚集环境的理化性质决定了tau菌株的构象，这可以使用拉曼光谱进行表征和区分。
Tauopathies are a group of disorders in which the deposition of abnormally folded tau protein accompanies neurodegeneration. The development of methods for detection and classification of pathological changes in protein conformation are desirable for understanding the factors that influence the structural polymorphism of aggregates in tauopathies. We have previously demonstrated the utility of Raman spectroscopy for the characterization and discrimination of different protein aggregates, including tau, based on their unique conformational signatures. Building on this, in the present study, we assess the utility of Raman spectroscopy for characterizing and distinguishing different conformers of the same protein which in the case of tau are unique tau strains generated in vitro. We now investigate the impact of aggregation environment, cofactors, post-translational modification and primary sequence on the Raman fingerprint of tau fibrils. Using quantitative conformational fingerprinting and multivariate statistical analysis, we found that the aggregation of tau in different buffer conditions resulted in the formation of distinct fibril strains. Unique spectral markers were identified for tau fibrils generated using heparin or RNA cofactors, as well as for phosphorylated tau. We also determined that the primary sequence of the tau monomer influenced the conformational signature of the resulting tau fibril, including 2N4R, 0N3R, K18 and P301S tau variants. These results highlight the conformational polymorphism of tau fibrils, which is reflected in the wide range of associated neurological disorders. Furthermore, the analyses presented in this study provide a benchmark for the Raman spectroscopic characterization of tau strains, which may shed light on how the aggregation environment, cofactors and post-translational modifications influence tau conformation in vivo in future studies. We report that the physiochemical properties of the aggregation environment dictate the conformation of tau strains, which can be characterized and distinguished using Raman spectroscopy.