Fluorous photoaffinity labeling to probe protein-small molecule interactions.
Fluorous photoaffinity labeling to probe protein-small molecule interactions.
复制标题
氟光亲和标记用于探测蛋白质-小分子相互作用。
DOI：
10.1007/978-1-4939-2269-7_20
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发表时间：
2015
期刊：
影响因子：
--
通讯作者：
Zhang,Qisheng
中科院分区：
文献类型：
--
作者：
Huang,Weigang;Zhang,Qisheng
识别生物活性小分子的细胞靶标对于它们作为化学探针或候选药物的应用是至关重要的。同样重要的是确定它们的“非靶标”相互作用，这通常是包括毒性在内的有害特性的原因。在分析小分子相互作用蛋白质的方法中，光亲和标记法因其独特的优点如灵敏度等而得到广泛应用。当与质谱学相结合时，这种方法可以提供额外的结构和机械信息，例如药物靶标的化学计量比和准确的相互作用氨基酸残基。我们描述了一种新的荧光光亲和标记方法，在光亲和标记试剂中加入一个氟标记，使标记的物种能够从复杂的混合物中浓缩用于分析。这一新特性可能使荧光光亲和标记方法适合于识别细胞环境中的瞬时相互作用以及低丰度、低亲和力的相互作用蛋白。
Identifying cellular targets of bioactive small molecules is essential for their applications as chemical probes or drug candidates. Of equal importance is to determine their “off-target” interactions, which usually account for unwanted properties including toxicity. Among strategies to profile small molecule-interacting proteins, photoaffinity labeling has been widely used because of its distinct advantages such as sensitivity. When combined with mass spectrometry, this approach can provide additional structural and mechanistic information, such as drug-target stoichiometry and exact interacting amino acid residues. We have described a novel fluorous photoaffinity labeling approach, in which a fluorous tag is incorporated into the photoaffinity labeling reagent to enable the enrichment of the labeled species from complex mixtures for analysis. This new feature likely makes the fluorous photoaffinity labeling approach suitable to identify transient interactions, and low-abundant, low-affinity interacting proteins in a cellular environment.