Page Annika C S, Orr Lauren M, Meyers Margot L, Belcher Bridget P, Coffey Theodore G, Scholz Spencer O, Cismoski Sabine, Nomura Daniel K, Toste F Dean
Department of Chemistry, University of California, Berkeley, Berkeley, CA 94720 USA.
bioRxiv. 2025 May 28:2025.05.24.655972. doi: 10.1101/2025.05.24.655972.
Deconvolution of the protein targets of hit compounds from phenotypic screens, often conducted in live cells, is critical for understanding mechanism of action and identifying potentially hazardous off-target interactions. While photoaffinity labeling and chemoproteomics are long-established approaches for discovering small-molecule-protein interactions in live cells, there are a relatively small number of photoaffinity labeling strategies that can be applied in intracellular settings. Recently, we reported a novel chemical framework for photoaffinity labeling based on the photo-Brook rearrangement of acyl silanes and demonstrated its ability, when appended to protein-targeting ligands, to label recombinant proteins. Here, we report the application of these probes to live cell photoaffinity workflows, demonstrate their complementarity to current state-of-the-art minimalist diazirine-based photoaffinity probes, and introduce a modular synthetic route to access acyl silane scaffolds with improved labeling properties.
从表型筛选中命中化合物的蛋白质靶点进行反卷积分析（通常在活细胞中进行），对于理解作用机制和识别潜在有害的脱靶相互作用至关重要。虽然光亲和标记和化学蛋白质组学是在活细胞中发现小分子-蛋白质相互作用的长期确立的方法，但可应用于细胞内环境的光亲和标记策略相对较少。最近，我们报道了一种基于酰基硅烷的光布鲁克重排的新型光亲和标记化学框架，并证明了其在连接到蛋白质靶向配体时标记重组蛋白的能力。在这里，我们报告了这些探针在活细胞光亲和工作流程中的应用，证明了它们与当前最先进的基于重氮环丙烷的极简光亲和探针的互补性，并介绍了一种模块化合成路线，以获得具有改进标记特性的酰基硅烷支架。
