Preparation of α-Synuclein Amyloid Assemblies for Toxicity Experiments
Preparation of α-Synuclein Amyloid Assemblies for Toxicity Experiments
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DOI：
10.1007/978-1-4939-7816-8_4
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发表时间：
2018-01-01
期刊：
影响因子：
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通讯作者：
Cremades, Nunilo
中科院分区：
文献类型：
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作者：
Chen, Serene W.;Cremades, Nunilo
某些蛋白质（包括帕金森病相关蛋白α-突触核蛋白）的淀粉样蛋白组装体通常与神经退行性疾病的发展和扩散相关，尽管最具毒性的形式的性质及其引发神经退行性疾病的机制在很大程度上仍然未知。这至少部分是由于制备可用于毒性实验的淀粉样蛋白组装体的稳定且结构均匀的样品所涉及的固有挑战。在这里，我们描述了两种不同类型的稳定的α-突触核蛋白淀粉样蛋白组件的制备，即动力学捕获的低聚物物种和繁殖能力的纤维状多晶型物。样品中的异质性程度已经被定义并仔细地最小化，从而允许在不同的α-突触核蛋白淀粉样蛋白组装体中建立有意义的结构-毒性关系。
Amyloid assemblies of certain proteins, including the Parkinson disease-related protein alpha-synuclein, are commonly associated with the development and spreading of neurodegenerative diseases, although the nature of the most toxic forms and the mechanisms by which they trigger neurodegeneration remain largely unknown. This is at least in part due to the inherent challenges involved in the preparation of stable and structurally homogeneous samples of amyloid assemblies that could be used in toxicity experiments. Here, we describe the preparation of two different types of stable alpha-synuclein amyloid assemblies, namely a kinetically trapped oligomeric species and a propagating-competent fibrillar polymorph. The degree of heterogeneity in the samples has been defined and carefully minimized, thus allowing for meaningful structure-toxicity relationships in different alpha-synuclein amyloid assemblies to be established.