State Key Laboratory for Mesoscopic Physics, School of Physics, Peking University, Beijing 100871, China.
Nanoscale. 2019 Mar 28;11(13):6480-6488. doi: 10.1039/c9nr00023b.
A30P and E46K are two mutants of α-synuclein (α-Syn) associated with familial early-onset Parkinson's disease (PD), and amyloid fibrils of α-Syn are the hallmarks of this disease. Detecting the heterogeneous system in the oligomerization stage of α-Syn is crucial for understanding the fibril formation and in vivo toxicity of α-Syn oligomers. Over the last two decades, solid-state nanopore technology has been developed into a reliable and versatile method in single-molecule studies. In this work, we study the time-dependent kinetics of early oligomerization of wild-type α-Syn, A30P, and E46K mutants through silicon nitride solid-state nanopores. By testing A30P, E46K, and wild-type α-Syn samples with different incubation times-from 3 to 15 days-we identify three typical types of oligomers formed in the oligomerization stage and confirm that A30P and E46K mutants aggregate faster than wild-type α-Syn. The results imply that the distinct aggregation pathways and kinetics featured by wild-type α-Syn and mutations may account for their distinct cytotoxicity and pathology in PD-related studies.
A30P 和 E46K 是与家族性早发性帕金森病（PD）相关的两种α-突触核蛋白（α-Syn）突变体，而α-Syn 的淀粉样纤维是这种疾病的标志。检测α-Syn 寡聚化阶段的异构体系对于理解纤维形成和体内α-Syn 寡聚物的毒性至关重要。在过去的二十年中，固态纳米孔技术已发展成为单分子研究中一种可靠且多功能的方法。在这项工作中，我们通过硅氮化物固态纳米孔研究了野生型α-Syn、A30P 和 E46K 突变体的早期寡聚化的时变动力学。通过测试具有不同孵育时间（3 至 15 天）的 A30P、E46K 和野生型α-Syn 样品，我们鉴定了在寡聚化阶段形成的三种典型寡聚物，并证实 A30P 和 E46K 突变体比野生型α-Syn 聚集得更快。结果表明，野生型α-Syn 和突变体的不同聚集途径和动力学特征可能解释了它们在 PD 相关研究中的不同细胞毒性和病理学。
