Department of Chemistry, East Carolina University, Greenville, North Carolina 27858, United States.
Department of Health and Human Services, Genome Integrity and Structural Biology Laboratory, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, North Carolina 27709, United States.
Biochemistry. 2022 Sep 6;61(17):1766-1773. doi: 10.1021/acs.biochem.2c00283. Epub 2022 Aug 24.
Accumulation of filamentous aggregates of α-synuclein is a pathological hallmark of several neurodegenerative diseases, including Parkinson's disease (PD). The interaction between α-synuclein and phospholipids has been shown to play a critical role in the aggregation of α-synuclein. Most structural studies have, however, been focused on α-synuclein filaments formed in the absence of lipids. Here, we report the structural investigation of α-synuclein filaments assembled under the quiescent condition in the presence of anionic lipid vesicles using electron microscopy (EM), including cryogenic electron microscopy (cryo-EM). Our transmission electron microscopy (TEM) analyses reveal that α-synuclein forms curly protofilaments at an early stage of aggregation. The flexible protofilaments were then converted to long filaments after a longer incubation of 30 days. More detailed structural analyses using cryo-EM reveal that the long filaments adopt untwisted structures with different diameters, which have not been observed in previous α-synuclein fibrils formed in vitro. The untwisted filaments are rather similar to straight filaments with no observable twist that are extracted from patients with dementia with Lewy bodies. Our structural studies highlight the conformational diversity of α-synuclein filaments, requiring additional structural investigation of not only more ex vivo α-synuclein filaments but also in vitro α-synuclein filaments formed in the presence of diverse cofactors to better understand the molecular basis of diverse molecular conformations of α-synuclein filaments.
α- 突触核蛋白丝状聚集体的积累是几种神经退行性疾病的病理标志,包括帕金森病(PD)。已经表明,α- 突触核蛋白与磷脂之间的相互作用在 α- 突触核蛋白的聚集过程中起着关键作用。然而,大多数结构研究都集中在没有脂质的情况下形成的 α- 突触核蛋白纤维上。在这里,我们报告了在存在阴离子脂质体的情况下,在安静条件下组装的 α- 突触核蛋白纤维的结构研究,包括低温电子显微镜(cryo-EM)。我们的透射电子显微镜(TEM)分析表明,α- 突触核蛋白在聚集的早期阶段形成卷曲原纤维。在更长时间(30 天)的孵育后,柔性原纤维被转化成长纤维。使用 cryo-EM 进行更详细的结构分析表明,长纤维采用不同直径的未扭曲结构,这在以前体外形成的 α- 突触核蛋白纤维中没有观察到。未扭曲的纤维与从具有Lewy 体痴呆的患者中提取的没有可观察到的扭曲的直纤维非常相似。我们的结构研究强调了 α- 突触核蛋白纤维的构象多样性,需要对不仅是更多的离体 α- 突触核蛋白纤维,而且还需要对存在不同共因子的体外 α- 突触核蛋白纤维进行额外的结构研究,以更好地理解 α- 突触核蛋白纤维的不同分子构象的分子基础。