Faculty of Science, NCCR Chemical Biology, University of Geneva, 30 Quai Ernest Ansermet, CH-1205 Geneva, Switzerland.
J Am Chem Soc. 2021 Nov 17;143(45):18932-18940. doi: 10.1021/jacs.1c07013. Epub 2021 Nov 5.
Stapled peptides with an enforced α-helical conformation have been shown to overcome major limitations in the development of short peptides targeting protein-protein interactions (PPIs). While the growing arsenal of methodologies to staple peptides facilitates their preparation, stapling methodologies are not broadly embraced in synthetic library screening. Herein, we report a strategy leveraged on hybridization of short PNA-peptide conjugates wherein nucleobase driven assembly facilitates ligation of peptide fragments and constrains the peptide's conformation into an α-helix. Using native chemical ligation, we show that a mixture of peptide fragments can be combinatorially ligated and used directly in affinity selection against a target of interest. This approach was exemplified with a focused library targeting the p-53/MDM2 interaction. One hundred peptides were obtained in a one-pot ligation reaction, selected by affinity against MDM2 immobilized on beads, and the best binders were identified by mass spectrometry.
具有强制α-螺旋构象的订书肽已被证明能够克服针对蛋白质-蛋白质相互作用（PPIs）的短肽开发中的主要限制。虽然将肽订书的方法越来越多，这使得它们的制备变得更加容易，但订书方法在合成文库筛选中并未得到广泛应用。在此，我们报告了一种利用短 PNA-肽缀合物杂交的策略，其中碱基驱动的组装促进肽片段的连接，并将肽的构象约束为α-螺旋。使用天然化学连接，我们表明可以组合连接肽片段混合物，并直接用于针对感兴趣的靶标的亲和选择。该方法通过针对 p-53/MDM2 相互作用的聚焦文库进行了示例。在一锅连接反应中获得了 100 个肽，通过与固定在珠上的 MDM2 的亲和力进行选择，并通过质谱鉴定出最佳结合物。
