A Photoaffinity Labeling-Based Chemoproteomics Strategy for Unbiased Target Deconvolution of Small Molecule Drug Candidates
A Photoaffinity Labeling-Based Chemoproteomics Strategy for Unbiased Target Deconvolution of Small Molecule Drug Candidates
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DOI：
10.1007/978-1-4939-7201-2_1
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发表时间：
2017-01-01
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影响因子：
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通讯作者：
Schirle, Markus
中科院分区：
文献类型：
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作者：
Thomas, Jason R.;Brittain, Scott M.;Schirle, Markus
光亲和标记（PAL）和定量化学蛋白质组学的结合，使全面，公正的蛋白质相互作用谱的测定，以支持生物活性小分子的目标识别。这种方法适用于培养中的细胞，并且与已知难以通过常规化学蛋白质组学方法获得的与细胞相关的跨膜靶类别（如G蛋白偶联受体和离子通道）相容。在这里，我们描述了一种策略，结合PAL探针滴定和竞争与过量的亲本化合物的目标，使识别特定的相互作用，以及评估的功能相关性的结合事件的表型正在调查。
The combination of photoaffinity labeling (PAL) and quantitative chemoproteomics enables the comprehensive, unbiased determination of protein interaction profiles to support target identification of bioactive small molecules. This approach is amenable to cells in culture and compatible with pharmacologically relevant transmembrane target classes like G-protein coupled receptors and ions channels which have been notoriously hard to access by conventional chemoproteomics approaches. Here, we describe a strategy that combines PAL probe titration and competition with excess parental compounds with the goal of enabling the identification of specific interactors as well as assessing the functional relevance of a binding event for the phenotype under investigation.