Schulich Faculty of Chemistry, Technion-Israel Institute of Technology, Haifa, 3200008, Israel.
Department of Chemistry, Graduate School of Science, The University of Tokyo, Bunkyo-ku, 7-3-1 Hongo, Bunkyo, Tokyo, 113-0033, Japan.
Nat Commun. 2022 Oct 18;13(1):6174. doi: 10.1038/s41467-022-33808-6.
Developing an effective binder for a specific ubiquitin (Ub) chain is a promising approach for modulating various biological processes with potential applications in drug discovery. Here, we combine the Random Non-standard Peptides Integrated Discovery (RaPID) method and chemical protein synthesis to screen an extended library of macrocyclic peptides against synthetic Lys63-linked Di-Ub to discover a specific binder for this Ub chain. Furthermore, next-generation binders are generated by chemical modifications. We show that our potent cyclic peptide is cell-permeable, and inhibits DNA damage repair, leading to apoptotic cell death. Concordantly, a pulldown experiment with the biotinylated analog of our lead cyclic peptide supports our findings. Collectively, we establish a powerful strategy for selective inhibition of protein-protein interactions associated with Lys63-linked Di-Ub using cyclic peptides. This study offers an advancement in modulating central Ub pathways and provides opportunities in drug discovery areas associated with Ub signaling.
开发针对特定泛素（Ub）链的有效结合物是一种很有前途的方法，可以用于调节各种生物过程，在药物发现方面具有潜在的应用价值。在这里，我们将随机非标准肽综合发现（RaPID）方法和化学蛋白质合成相结合，针对合成的 Lys63 连接的二泛素筛选了一个扩展的大环肽文库，以发现针对这种 Ub 链的特定结合物。此外，通过化学修饰生成了下一代结合物。我们表明，我们的有效环状肽具有细胞通透性，并抑制 DNA 损伤修复，导致细胞凋亡。相应地，用我们的先导环状肽的生物素化类似物进行的下拉实验支持了我们的发现。总的来说，我们建立了一种使用环状肽选择性抑制与 Lys63 连接的二泛素相关的蛋白质-蛋白质相互作用的强大策略。这项研究为调节中央 Ub 途径提供了新的进展，并为 Ub 信号相关的药物发现领域提供了机会。
