Aggregation and the formation of oligomeric intermediates of amyloid-β (Aβ) at the membrane interface of neuronal cells are implicated in the cellular toxicity and pathology of Alzheimer’s disease. Small molecule compounds have been shown to suppress amyloid aggregation and cellular toxicity, but often the presence of a lipid membrane negates their activity. A high-throughput screen of 1800 small molecules was performed to search for membrane active inhibitors, and 21 primary hits were discovered. Through the use of fluorescence-based assays, transmission electron microscopy, and dot blot assays, the initial 21 primary hits were narrowed down to five lead compounds. Nuclear magnetic resonance and circular dichroism experiments were used for further confirmation of amyloid inhibition at the membrane interface and to obtain insights into the secondary structure of amyloid-β, while size exclusion chromatography was used to characterize the size of Aβ species. Lastly, dye-leakage assays allowed us to understand how the addition of the five lead compounds affected amyloid-β’s ability to permeate the lipid bilayer. These results provide insights into small molecules that stabilize small amyloid species in the presence of membranes for the development of tool compounds for deeper investigations of these transient species.

中文翻译：

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膜界面的高通量筛选揭示了β淀粉样蛋白的抑制剂。

神经元细胞膜界面上淀粉样蛋白-β (Aβ) 寡聚中间体的聚集和形成与阿尔茨海默病的细胞毒性和病理学有关。小分子化合物已被证明可以抑制淀粉样蛋白聚集和细胞毒性，但脂质膜的存在通常会抵消它们的活性。对 1800 个小分子进行了高通量筛选，以寻找膜活性抑制剂，并发现了 21 个主要目标。通过使用基于荧光的测定、透射电子显微镜和斑点印迹测定，最初的 21 种主要化合物被缩小到 5 种先导化合物。核磁共振和圆二色性实验用于进一步确认膜界面上淀粉样蛋白的抑制，并深入了解淀粉样蛋白-β的二级结构，同时使用尺寸排阻色谱法来表征Aβ物种的尺寸。最后，染料渗漏分析使我们能够了解五种先导化合物的添加如何影响淀粉样蛋白-β 渗透脂质双层的能力。这些结果提供了对在膜存在下稳定小淀粉样蛋白种类的小分子的见解，以开发工具化合物来更深入地研究这些瞬态种类。染料渗漏分析使我们能够了解五种先导化合物的添加如何影响淀粉样蛋白-β 渗透脂质双层的能力。这些结果提供了对在膜存在下稳定小淀粉样蛋白种类的小分子的见解，以开发工具化合物来更深入地研究这些瞬态种类。染料渗漏分析使我们能够了解五种先导化合物的添加如何影响淀粉样蛋白-β 渗透脂质双层的能力。这些结果提供了对在膜存在下稳定小淀粉样蛋白种类的小分子的见解，以开发工具化合物来更深入地研究这些瞬态种类。