Targeting UBE2C for degradation by bioPROTACs based on bacterial E3 ligase
Targeting UBE2C for degradation by bioPROTACs based on bacterial E3 ligase
复制标题
基于细菌 E3 连接酶的 bioPROTAC 靶向 UBE2C 进行降解
DOI：
10.1016/j.cclet.2022.08.012
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发表时间：
2023-01-17
影响因子：
9.1
通讯作者：
Ye，Yuxin
中科院分区：
文献类型：
--
作者：
Wang，Jinpeng;Zhang，Min;Ye，Yuxin
UBE2C(泛素结合酶E2 C)是细胞周期进程的关键调控因子，是发现抗肿瘤药物的重要靶点。然而，由于UBE2C缺乏“可下药”的口袋，开发UBE2C的抑制剂是具有挑战性的。生物蛋白水解靶向嵌合体是一种以蛋白质为基础的降解物，通过将接头融合到E3连接酶的一个亚基上，进行泛素化，随后依赖蛋白酶体降解目标蛋白。我们报道了基于细菌E3连接酶IpaH9.8的NEL(新的E3连接酶)结构域和APC2(后期促进复合体2)的UBE2C结合的WHB(有翼螺旋B)结构域的UBE2C靶向生物PROTAC的设计和生物学评价。玻尿素化试验和质谱分析表明，该生物PROTAC能将泛素转移到UBE2C表面暴露的赖氨酸上，并催化形成多聚泛素链。此外，瞬时共表达实验表明，生物PROTAC能够促进异源UBE2C蛋白酶体的降解，挽救其下游底物。
UBE2C (Ubiquitin conjugating enzyme E2 C), a key regulator of cell cycle progression, is a promising target for discovery of antitumor agents. However, it is challenging to develop inhibitors of UBE2C owing to its lack of “druggable” pockets. BioPROTACs (biological proteolysis targeting chimeras) are a kind of protein-based degraders by fusing an adaptor to a subunit of E3 ligase for ubiquitination and subsequent proteasome-dependent degradation of target protein. We report herein the design and biological evaluation of a UBE2C-targeting bioPROTAC based on the NEL (novel E3 ligase) domain of bacterial E3 ligase IpaH9.8 and the UBE2C-binding WHB (winged-helix B) domain of APC2 (anaphase promoting complex subunit 2). Thein vitroubiquitination test and Mass Spectrometry analysis showed that the bioPROTAC could transfer ubiquitin to surface exposed lysines on UBE2C and catalyzed the formation of polyubiquitin chains. In addition, the transient co-expression experiment showed that the bioPROTAC could promote proteasomal degradation of heterologous UBE2C and rescue its downstream substrates in mammalian cells.