Preparation of stable amyloid β-protein oligomers of defined assembly order.
Preparation of stable amyloid β-protein oligomers of defined assembly order.
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DOI：
10.1007/978-1-61779-551-0_3
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发表时间：
2012
期刊：
影响因子：
--
通讯作者：
Teplow DB
中科院分区：
文献类型：
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作者：
Rosensweig C;Ono K;Murakami K;Lowenstein DK;Bitan G;Teplow DB
淀粉样β蛋白（Aβ）的寡聚体被认为是阿尔茨海默病（AD）中最接近的神经毒性物质。低聚物的形成是一个复杂的过程，产生亚稳结构的多分散群体。由于这个原因，正式的结构-活性相关性，在体外和体内，一直难以实现。通过将光化学交联方法应用于Aβ组装体系，提供了该问题的解析解。这种方法，光诱导交联未修饰的蛋白质（PICUP），使定量测定的寡聚体的大小分布。我们在这里报告的集成PICUP与SDS-PAGE和碱提取程序，以创建一个方法，用于分离的纯群体的低聚物的定义的顺序。该方法已成功用于为Aβ低聚物的正式结构-活性研究提供材料。
Oligomeric assemblies of the amyloid β-protein, Aβ, are thought to be the proximate neurotoxic agents in Alzheimer’s disease (AD). Oligomer formation is a complex process that produces a polydisperse population of metastable structures. For this reason, formal structure–activity correlations, both in vitro and in vivo, have been difficult to accomplish. An analytical solution to this problem was provided by the application of a photochemical cross-linking method to the Aβ assembly system. This method, photo-induced cross-linking of unmodified proteins (PICUP), enabled the quantitative determination of the oligomer size distribution. We report here the integration of PICUP with SDS-PAGE and alkaline extraction procedures to create a method for the isolation of pure populations of oligomers of defined order. This method has been used successfully to provide material for formal structure–activity studies of Aβ oligomers.