Characterization of chemical libraries for luciferase inhibitory activity
Characterization of chemical libraries for luciferase inhibitory activity
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DOI：
10.1021/jm701302v
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发表时间：
2008-04-24
影响因子：
7.3
通讯作者：
Inglese, James
中科院分区：
文献类型：
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作者：
Auld, Douglas S.;Southall, Noel T.;Inglese, James
为了帮助解释基于荧光素酶测定的高通量筛选（HTS）结果，我们使用了定量HTS，该方法定义了每个库样品的浓度 - 反应行为，以介绍来自Photinus pyralis的ATP依赖性荧光素酶针对超过7万个样本。我们发现，大约3％的文库是活性的，仅包含具有抑制浓度的化合物，其中681（0.9％）（0.9％）表现出IC50 <10 MU M.代表性化合物均显示出抑制纯化的P. pyralis以及几个商业性的。基于荧光素酶的检测试剂，但被发现对肾肾形荧光素酶的不活跃。还检查了样品的光衰减，发现在肾叶肠杆菌荧光素酶产生的蓝移生物发光中比在Pyralis luciferase产生的生物发光中更为突出。我们描述了荧光素酶抑制剂的结构 - 活性关系，并讨论了该数据在HTS结果解释和基于荧光素酶测定的配置中的使用。
To aid in the interpretation of high-throughput screening (HTS) results derived from luciferase-based assays, we used quantitative HTS, an approach that defines the concentration -response behavior of each library sample, to profile the ATP-dependent luciferase from Photinus pyralis against more than 70 000 samples. We found that approximately 3% of the library was active, containing only compounds with inhibitory concentration -responses, of which 681 (0.9%) exhibited IC50 < 10 mu M. Representative compounds were shown to inhibit purified P. pyralis as well as several commercial luciferase-based detection reagents but were found to be largely inactive against Renilla reniformis luciferase. Light attenuation by the samples was also examined and found to be more prominent in the blue-shifted bioluminescence produced by R. reniformis luciferase than in the bioluminescence produced by P. pyralis luciferase. We describe the structure- activity relationship of the luciferase inhibitors and discuss the use of this data in the interpretation of HTS results and configuration of luciferase-based assays.