Department of Medicinal Chemistry and Molecular Pharmacology , Purdue Center for Cancer Research, Purdue University , West Lafayette , Indiana 47907 , United States.
J Am Chem Soc. 2019 Oct 30;141(43):17057-17061. doi: 10.1021/jacs.9b08085. Epub 2019 Oct 22.
We report the selection of DNA-encoded small molecule libraries against protein targets within the cytosol and on the surface of live cells. The approach relies on generation of a covalent linkage of the DNA to protein targets by affinity labeling. This cross-linking event enables subsequent copurification by a tag on the recombinant protein. To access targets within cells, a cyclic cell-penetrating peptide is appended to DNA-encoded libraries for delivery across the cell membrane. As this approach assesses binding of DELs to targets in live cells, it provides a strategy for selection of DELs against challenging targets that cannot be expressed and purified as active.
我们报告了针对细胞质内和活细胞表面的蛋白质靶标进行 DNA 编码小分子文库筛选的方法。该方法依赖于通过亲和标记将 DNA 与蛋白质靶标共价连接。这种交联事件可通过重组蛋白上的标签进行后续共纯化。为了访问细胞内的靶标，将环状细胞穿透肽附加到 DNA 编码文库上，以穿过细胞膜进行递呈。由于这种方法评估了 DEL 在活细胞中与靶标的结合，因此为选择针对难以表达和纯化为活性形式的靶标的 DEL 提供了一种策略。
