Allerton Siân C, Kuimova Marina K, Aprile Francesco A
Department of Chemistry, Molecular Sciences Research Hub, Imperial College London, London W12 0BZ, U.K.
Institute of Chemical Biology, Molecular Sciences Research Hub, Imperial College London, London W12 0BZ, U.K.
ACS Appl Mater Interfaces. 2025 Feb 19;17(7):10499-10508. doi: 10.1021/acsami.4c21710. Epub 2025 Feb 5.
α-Synuclein is an intrinsically disordered protein that forms amyloids in Parkinson's disease. Currently, detection methods predominantly report on the formation of mature amyloids but are weakly sensitive to the early stage, toxic oligomers. Molecular rotors are fluorophores that sense changes in the viscosity of their local environment. Here, we monitor α-synuclein oligomer formation using the fluorescence lifetime of molecular rotors. We detect oligomer formation and conversion into amyloids for wild-type and two α-synuclein variants, the pathological mutant A30P and ΔP1 α-synuclein, which lacks a master regulator region of aggregation (residues 36-42). We report that A30P α-synuclein shows a rate of oligomer formation similar to that of wild-type α-synuclein, whereas ΔP1 α-synuclein shows delayed oligomer formation. Additionally, both variants demonstrate a slower conversion of oligomers into amyloids. Our method provides a quantitative approach to unveiling the complex mechanism of α-synuclein aggregation, which is key to understanding the pathology of Parkinson's disease.
α-突触核蛋白是一种内在无序的蛋白质，在帕金森病中会形成淀粉样蛋白。目前，检测方法主要报告成熟淀粉样蛋白的形成情况，但对早期有毒寡聚体的敏感性较弱。分子转子是一种荧光团，可感知其局部环境粘度的变化。在此，我们利用分子转子的荧光寿命监测α-突触核蛋白寡聚体的形成。我们检测了野生型以及两种α-突触核蛋白变体（病理性突变体A30P和缺少聚集主调控区域（第36 - 42位氨基酸残基）的ΔP1 α-突触核蛋白）的寡聚体形成及向淀粉样蛋白的转化。我们报告称，A30P α-突触核蛋白的寡聚体形成速率与野生型α-突触核蛋白相似，而ΔP1 α-突触核蛋白的寡聚体形成延迟。此外，两种变体的寡聚体向淀粉样蛋白的转化都较慢。我们的方法提供了一种定量方法来揭示α-突触核蛋白聚集的复杂机制，这对于理解帕金森病的病理学至关重要。
