Xanthine-based photoaffinity probes allow assessment of ligand engagement by TRPC5 channels
Xanthine-based photoaffinity probes allow assessment of ligand engagement by TRPC5 channels
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DOI：
10.26434/chemrxiv.11890128.v1
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发表时间：
2020-02
影响因子：
4.1
通讯作者：
C. C. Bauer-C.;Aisling Minard;Isabelle B. Pickles;K. Simmons;Eulashini Chuntharpursat-Bon;M. Burnham;
中科院分区：
文献类型：
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作者：
C. C. Bauer-C.;Aisling Minard;Isabelle B. Pickles;K. Simmons;Eulashini Chuntharpursat-Bon;M. Burnham;
TRPC 1/4/5阳离子通道是用于治疗中枢神经系统（CNS）病症、肾脏疾病以及心血管和代谢疾病等的新兴药物靶标。已经报道了各种小分子TRPC 1/4/5调节剂，包括区分特定TRPC 1/4/5四聚体的高效黄嘌呤衍生物。然而，缺乏在活细胞中通过TRPC 1/4/5通道分析配体接合的工具。在这里，我们报告了一组有效的基于黄嘌呤的光亲和探针，功能上模仿黄嘌呤Pico 145和AM 237。使用这些探针，我们已经开发了一个光亲和标记协议的TRPC 5通道，提供了第一种方法的定量评估TRPC 5与细胞中的小分子的结合相互作用。这种方法对于靶向TRPC 1/4/5通道的黄嘌呤/脂质结合位点的药物发现工作可能是重要的。
TRPC1/4/5 cation channels are emerging drug targets for the treatment of, amongst others, central nervous system (CNS) disorders, kidney disease, and cardiovascular and metabolic disease. Various small-molecule TRPC1/4/5 modulators have been reported, including highly potent xanthine derivatives that distinguish between specific TRPC1/4/5 tetramers. However, tools to profile ligand engagement by TRPC1/4/5 channels in live cells are lacking. Here, we report a set of potent xanthine-based photoaffinity probes that functionally mimic the xanthines Pico145 and AM237. Using these probes, we have developed a photoaffinity labelling protocol for TRPC5 channels, providing the first method for the quantitative assessment of binding interactions of TRPC5 with small molecules in cells. This method could be important for drug discovery efforts targeting the xanthine/lipid binding site of TRPC1/4/5 channels.