三元络合物
三元运算
降级(电信)
化学
生化工程
计算机科学
工程类
生物化学
酶
电信
程序设计语言
作者
Martin P. Schwalm,Andreas Krämer,Anja Dölle,Janik Weckesser,Xufen Yu,Jian Jin,Krishna Saxena,Stefan Knapp
标识
DOI:10.1016/j.chembiol.2023.06.002
摘要
The multi-step degradation process of PROteolysis TArgeting Chimeras (PROTACs) poses a challenge for their rational development, as the rate-limiting steps that determine PROTACs efficiency remain largely unknown. Moreover, the slow throughput of currently used endpoint assays does not allow the comprehensive analysis of larger series of PROTACs. Here, we developed cell-based assays using the NanoLuciferase and HaloTag that allow measuring PROTAC-induced degradation and ternary complex formation kinetics and stability in cells. Using PROTACs developed for the degradation of WD40 repeat domain protein 5 (WDR5), the characterization of the mode of action of these PROTACs in the early degradation cascade revealed a key role of ternary complex formation and stability. Comparing a series of ternary complex crystal structures highlighted the importance of an efficient E3-target interface for ternary complex stability. The developed assays outline a strategy for the rational optimization of PROTACs using a series of live cell assays monitoring key steps of the early PROTAC-induced degradation pathway.
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