Department of Chemistry and State Key Laboratory of Synthetic Chemistry, The University of Hong Kong, Hong Kong SAR, China.
Chongqing Key Laboratory of Natural Product Synthesis and Drug Research, School of Pharmaceutical Sciences, Chongqing University, Chongqing, China.
Nat Chem. 2021 Jan;13(1):77-88. doi: 10.1038/s41557-020-00605-x. Epub 2020 Dec 21.
Membrane proteins on the cell surface perform a myriad of biological functions; however, ligand discovery for membrane proteins is highly challenging, because a natural cellular environment is often necessary to maintain protein structure and function. DNA-encoded chemical libraries (DELs) have emerged as a powerful technology for ligand discovery, but they are mainly limited to purified proteins. Here we report a method that can specifically label membrane proteins with a DNA tag, and thereby enable target-specific DEL selections against endogenous membrane proteins on live cells without overexpression or any other genetic manipulation. We demonstrate the generality and performance of this method by screening a 30.42-million-compound DEL against the folate receptor, carbonic anhydrase 12 and the epidermal growth factor receptor on live cells, and identify and validate a series of novel ligands for these targets. Given the high therapeutic significance of membrane proteins and their intractability to traditional high-throughput screening approaches, this method has the potential to facilitate membrane-protein-based drug discovery by harnessing the power of DEL.
细胞膜表面的膜蛋白具有多种生物学功能；然而，由于天然细胞环境对于维持蛋白质结构和功能至关重要，因此膜蛋白的配体发现极具挑战性。DNA 编码的化学文库 (DEL) 的出现为配体发现提供了一种强大的技术，但它们主要局限于纯化蛋白。在这里，我们报告了一种可以用 DNA 标签特异性标记膜蛋白的方法，从而能够在不进行过表达或任何其他遗传操作的情况下，针对活细胞内的内源性膜蛋白进行靶向特异性 DEL 选择。我们通过在活细胞上针对叶酸受体、碳酸酐酶 12 和表皮生长因子受体筛选了一个包含 3042 万个化合物的 DEL，证明了该方法的通用性和性能，并鉴定和验证了针对这些靶标的一系列新型配体。鉴于膜蛋白的高治疗意义及其对传统高通量筛选方法的不可操作性，该方法有可能通过利用 DEL 的力量来促进基于膜蛋白的药物发现。
