Proteome Dynamics Lab, Max Delbruck Center for Molecular Medicine in the Helmholtz Association, Robert-Roessle-Str. 10, 13125, Berlin, Germany.
Medicinal Chemistry, Leibniz-Forschungsinstitut für Molekulare Pharmakologie (FMP), 13125, Berlin, Germany.
ChemMedChem. 2022 Oct 6;17(19):e202200346. doi: 10.1002/cmdc.202200346. Epub 2022 Sep 20.
Identifying the protein targets of drugs is an important but tedious process. Existing proteomic approaches enable unbiased target identification but lack the throughput needed to screen larger compound libraries. Here, we present a compound interaction screen on a photoactivatable cellulose membrane (CISCM) that enables target identification of several drugs in parallel. To this end, we use diazirine-based undirected photoaffinity labeling (PAL) to immobilize compounds on cellulose membranes. Functionalized membranes are then incubated with protein extract and specific targets are identified via quantitative affinity purification and mass spectrometry. CISCM reliably identifies known targets of natural products in less than three hours of analysis time per compound. In summary, we show that combining undirected photoimmobilization of compounds on cellulose with quantitative interaction proteomics provides an efficient means to identify the targets of natural products.
鉴定药物的蛋白质靶标是一个重要但繁琐的过程。现有的蛋白质组学方法能够实现无偏靶标鉴定，但缺乏筛选更大化合物库所需的通量。在这里，我们提出了一种在光活化纤维素膜（CISCM）上进行的化合物相互作用筛选方法，能够并行鉴定几种药物的靶标。为此，我们使用基于重氮化物的无导向光亲和标记（PAL）将化合物固定在纤维素膜上。然后将功能化的膜与蛋白质提取物一起孵育，并通过定量亲和纯化和质谱鉴定特定的靶标。CISCM 能够可靠地在不到三小时的分析时间内鉴定每种化合物的天然产物的已知靶标。总之，我们表明，将化合物在纤维素上的无导向光固定与定量相互作用蛋白质组学相结合，为鉴定天然产物的靶标提供了一种有效的手段。
