Multiple-State Monitoring of SOD1 Amyloid Formation at Single-Residue Resolution by Rheo-NMR Spectroscopy
Multiple-State Monitoring of SOD1 Amyloid Formation at Single-Residue Resolution by Rheo-NMR Spectroscopy
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DOI：
10.1021/jacs.1c02974
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发表时间：
2021-07-07
影响因子：
15
通讯作者：
Sugase, Kenji
中科院分区：
文献类型：
--
作者：
Iwakawa, Naoto;Morimoto, Daichi;Sugase, Kenji
蛋白质聚集体或原纤维的形成需要可溶性天然蛋白质单体通过多种分子状态的转化。没有光谱技术已经成功地捕捉到了瞬间分子尺度的原纤化事件。在这里，我们报告残留物和状态特定的实时监测肌萎缩侧索硬化症相关的SOD 1的纤颤流变NMR（Rheo-NMR）光谱。在中度变性条件下，折叠和未折叠的单体SOD1的NMR信号是同时可观察到的，折叠的单体SOD1的交叉峰强度下降速度比未折叠的物种，并在折叠的SOD1的310-螺旋变形之前，全球展开。此外，实时蛋白质动力学分析确定了参与SOD 1寡聚体核心结构形成的残基。我们的研究结果提供了深入了解SOD1和原纤维形成中的局部和全局展开事件。这种流变核磁共振分析将不仅适用于其他淀粉样蛋白质的原子水平的监测，但也定量剪切诱导的结构变化的非淀粉样蛋白质和阐明剪切增强的化学现象，如粘度增加和结晶的各种溶液状态的化合物。
Formation of protein aggregates or fibrils entails the conversion of soluble native protein monomers via multiple molecular states. No spectroscopic techniques have succeeded in capturing the transient molecular-scale events of fibrillation in situ. Here we report residue- and state-specific real-time monitoring of the fibrillation of amyotrophic lateral sclerosis-related SOD1 by rheology NMR (Rheo-NMR) spectroscopy. Under moderately denaturing conditions, where NMR signals of folded and unfolded monomeric SOD1 are simultaneously observable, the cross-peak intensities of folded monomeric SOD1 decreased faster than those of the unfolded species, and a 310-helix in folded SOD1 was deformed prior to global unfolding. Furthermore, real-time protein dynamics analysis identified residues involved in the core structure formation of SOD1 oligomers. Our findings provide insight into local and global unfolding events in SOD1 and fibril formation. This Rheo-NMR analysis will be applicable not only to atomic-level monitoring of other amyloidogenic proteins but also to quantification of shear-induced structural changes of non-amyloidogenic proteins and elucidation of shear-enhanced chemical phenomena such as viscosity increase and crystallization of various solution-state compounds.