A high-throughput screen for compounds that inhibit aggregation of the Alzheimer's peptide
A high-throughput screen for compounds that inhibit aggregation of the Alzheimer's peptide
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DOI：
10.1021/cb600135w
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发表时间：
2006-01-01
影响因子：
4
通讯作者：
Hecht, Michael H.
中科院分区：
文献类型：
--
作者：
Kim, Woojin;Kim, Yunkyoung;Hecht, Michael H.
阿尔茨海默氏症多肽Aβ的聚集产生了有毒的多聚体，这些多聚体在阿尔茨海默病的发展中起着关键作用。抑制这种聚集的化合物可能被证明是预防或治疗阿尔茨海默病的有效治疗剂。虽然聚集抑制剂可能已经存在于组合文库中，但以经济高效的高通量方式找到这些化合物是一个巨大的挑战。为了迎接这一挑战，我们开发了一种新的高通量筛选，能够从大量非活性候选者库中分离Aβ聚集的抑制剂。这款屏幕使用了A beta 42和GFP的融合。在没有抑制的情况下，Aβ42的快速错误折叠和聚集导致整个融合蛋白错误折叠，从而阻止荧光。抑制Aβ42聚集的化合物使GFP能够折叠成其自然结构，并通过产生的荧光信号进行识别。通过对三嗪类化合物的中试文库进行筛选，我们已经确定了几种假定的抑制剂。通过一系列生化和生物物理方法对其中一种化合物进行了详细的研究。这些研究证实，所选择的化合物抑制合成的Aβ42肽的聚集。这里描述的基于荧光的方法是快速和廉价的，可以用于筛选Aβ42聚集和/或淀粉样蛋白生成抑制物的大型文库。
Aggregation of the Alzheimer's peptide A beta produces toxic multimeric species that play a key role in the development of Alzheimer's disease. Compounds that inhibit this aggregation may prove useful as therapeutic agents for the prevention or treatment of Alzheimer's disease. Although aggregation inhibitors may already exist in combinatorial libraries, finding these compounds in a cost-effective high-throughput manner poses an enormous challenge. To meet this challenge, we have developed a novel high-throughput screen capable of isolating inhibitors of A beta aggregation from large libraries of inactive candidates. The screen uses a fusion of A beta 42 to GFP. In the absence of inhibition, the rapid misfolding and aggregation of A beta 42 causes the entire fusion protein to misfold, thereby preventing fluorescence. Compounds that inhibit A beta 42 aggregation enable GFP to fold into its native structure and be identified by the resulting fluorescent signal. By implementing the screen on a pilot library of triazine derivatives, we have identified several putative inhibitors. One of the selected compounds was studied in detail by a series of biochemical and biophysical methods. These studies confirmed that the selected compound inhibits aggregation of synthetic A beta 42 peptide. The fluorescence-based method described here is rapid and inexpensive and can be used to screen large libraries for inhibitors of A beta 42 aggregation and/or amyloidogenesis.