Universal screening methods and applications of ThermoFluor®
Universal screening methods and applications of ThermoFluor®
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DOI：
10.1177/1087057106292746
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发表时间：
2006-10-01
影响因子：
--
通讯作者：
Nelen, Marina I.
中科院分区：
文献类型：
--
作者：
Cummings, Maxwell D.;Farnum, Michael A.;Nelen, Marina I.
基因组学革命揭示了大量特征不明确的蛋白质。科学家们经常能够制造出毫克级的蛋白质，而这些蛋白质的功能是未知的或假设的，仅基于非常遥远的序列同源性。广泛适用的功能表征工具对于阐明这些孤儿蛋白至关重要。另一个挑战是直接检测蛋白质-蛋白质相互作用的抑制剂（和变构效应）。这两个研究问题都与寻找和验证药物作用的新蛋白质靶点的挑战有关。长期以来，制药行业一直将筛选小分子集合作为发现药物先导的一种方法。在这种情况下，筛选通常涉及基于功能的分析。给定足够数量的感兴趣蛋白质，可能仍然需要大量的努力来进行功能表征，分析开发和分析配置以进行筛选。越来越多的技术报道有助于筛选功能未知的蛋白质的特定配体。这样的技术也允许功能独立筛选更好地表征蛋白质。ThermoFluor (R)是一种基于监测配体对温度依赖性蛋白质展开作用的筛选仪器，可在蛋白质功能未知的情况下应用。这项技术已被证明在解密一种重要的细菌酶和发现一系列癌症相关的蛋白质-蛋白质相互作用的抑制剂方面是有用的。作者回顾了药物发现中与这些研究问题相关的一些工具，并描述了我们对两种不同蛋白质的经验。
The genomics revolution has unveiled a wealth of poorly characterized proteins. Scientists are often able to produce milligram quantities of proteins for which function is unknown or hypothetical, based only on very distant sequence homology. Broadly applicable tools for functional characterization are essential to the illumination of these orphan proteins. An additional challenge is the direct detection of inhibitors of protein-protein interactions (and allosteric effectors). Both of these research problems are relevant to, among other things, the challenge of finding and validating new protein targets for drug action. Screening collections of small molecules has long been used in the pharmaceutical industry as 1 method of discovering drug leads. Screening in this context typically involves a function-based assay. Given a sufficient quantity of a protein of interest, significant effort may still be required for functional characterization, assay development, and assay configuration for screening. Increasingly, techniques are being reported that facilitate screening for specific ligands for a protein of unknown function. Such techniques also allow for function-independent screening with better characterized proteins. ThermoFluor (R), a screening instrument based on monitoring ligand effects on temperature-dependent protein unfolding, can be applied when protein function is unknown. This technology has proven useful in the decryption of an essential bacterial enzyme and in the discovery of a series of inhibitors of a cancer-related, protein-protein interaction. The authors review some of the tools relevant to these research problems in drug discovery, and describe our experiences with 2 different proteins.