A fluorescence polarization assay for inhibitors of Hsp90
A fluorescence polarization assay for inhibitors of Hsp90
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DOI：
10.1016/j.ab.2005.12.023
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发表时间：
2006-03-15
影响因子：
2.9
通讯作者：
Drysdale, MJ
中科院分区：
文献类型：
--
作者：
Howes, R;Barril, X;Drysdale, MJ
Hsp90 编码一种在物种间保守的普遍存在的分子伴侣蛋白，作用于多种底物，其中许多底物是重要的细胞信号蛋白。 Hsp90 功能的抑制已被推广为一种降解参与肿瘤发生和疾病进展的客户蛋白的机制。目前存在几种监测 Hsp90 功能抑制的测定方法，但在药物发现活动中的使用受到限制。利用初始命中化合物的晶体结构数据，我们开发了一种荧光偏振测定法来监测化合物与 Hsp90 的 ATP 结合位点的结合。该测定非常稳健 (Z' > 0.9)，可以检测 IC(50) 达 40 nM 的化合物的亲和力。我们将此测定法与小分子的共晶结构结合使用，以驱动基于结构的设计程序，旨在发现和优化一类新型有效的 Hsp90 抑制剂。 (c) 2006 Elsevier Inc. 保留所有权利。
Hsp90 encodes a ubiquitous molecular chaperone protein conserved among species which acts on multiple substrates, many of which are important cell-signaling proteins. Inhibition of Hsp90 function has been promoted as a mechanism to degrade client proteins involved in tumorigenesis and disease progression. Several assays to monitor inhibition of Hsp90 function currently exist but are limited in their use for a drug discovery campaign. Using data from the crystal structure of an initial hit compound, we have developed a fluorescence polarization assay to monitor binding of compounds to the ATP-binding site of Hsp90. This assay is very robust (Z' > 0.9) and can detect affinity of compounds with IC(50)s to 40 nM. We have used this assay in conjunction with cocrystal structures of small molecules to drive a structure-based design program aimed at the discovery and optimization of a novel class of potent Hsp90 inhibitors. (c) 2006 Elsevier Inc. All rights reserved.