Development and implementation of a miniaturized high-throughput time-resolved fluorescence energy transfer assay to identify small molecule inhibitors of polo-like kinase 1.
Development and implementation of a miniaturized high-throughput time-resolved fluorescence energy transfer assay to identify small molecule inhibitors of polo-like kinase 1.
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开发和实施小型化高通量时间分辨荧光能量转移测定，以鉴定 polo 样激酶 1 的小分子抑制剂。
DOI：
10.1089/adt.2007.102
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发表时间：
2007
影响因子：
1.8
通讯作者：
Lazo,JohnS
中科院分区：
文献类型：
--
作者：
Sharlow,ElizabethR;Leimgruber,Stephanie;Shun,TongYing;Lazo,JohnS
Polo-like kinase (Plk) 1 is a key enzyme involved in regulating the mammalian cell cycle that is also a validated anticancer drug target. Nonetheless, there are relatively few readily available potent and selective small molecule inhibitors of Plk1. To increase the availability of pharmacologically valuable Plk1 inhibitors, we describe herein the development, variability assessment, validation, and implementation of a 384-well automated, miniaturized high-throughput time-resolved fluorescence energy transfer screening assay designed to identify Plk1 kinase inhibitors. Using a small molecule library of pharmaceutically active compounds to gauge high-throughput assay robustness and reproducibility, we found nine general kinase inhibitors, including H-89, which was selected as the minimum control. We then interrogated a 97,101 compound library from the National Institutes of Health repository for small molecule inhibitors of Plk1 kinase activity. The initial primary hit rate in a single 10μMconcentration format was 0.21%. Hit compounds were subjected to concentration–response confirmation and interference assays. Identified in the screen were seven compounds with 50% inhibitory concentration (IC50) values below 1μM, 20 compounds with IC50values between 1μMand 5μM, and eight compounds with IC50values between 5 and 10μM, which could be assigned to seven distinct chemotype classes. Hit compounds were also examined for their ability to inhibit other kinases such as protein kinase D, focal adhesion kinase, rho-associated coiled coil protein kinase 2, c-jun NH2-terminal kinase 3, and protein kinase A via experimentation or data-mining. These compounds should be useful as probes for the biological activity of Plk1 and as leads for the development of new selective inhibitors of Plk1.
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影响因子：
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通讯作者：
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影响因子：
1.8
作者：
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通讯作者：
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DOI：
--
发表时间：
2007
期刊：
Biochemical and Biophysical Research Communications - BBRC
影响因子：
--
作者：
M. Wada;Y. Kawahito;S. Kimura;M. Kohno;H. Ishino;Mizuho Kimura;A. Omoto;A. Yamamoto;M. Hamaguchi;Y. Tsubouchi;D. Tokunaga;T. Hojo;E. Ashihara;T. Maekawa;T. Yoshikawa
通讯作者：
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