An E1-Catalyzed Chemoenzymatic Strategy to Isopeptide-N-Ethylated Deubiquitylase-Resistant Ubiquitin Probes
An E1-Catalyzed Chemoenzymatic Strategy to Isopeptide-N-Ethylated Deubiquitylase-Resistant Ubiquitin Probes
复制标题
E1 催化的抗异肽-N-乙基化去泛素化酶泛素探针的化学酶策略
DOI：
10.1002/anie.202002974
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发表时间：
2020
期刊：
影响因子：
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通讯作者：
Liu Lei
中科院分区：
文献类型：
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作者：
Zheng Qingyun;Wang Tian;Chu Guo-Chao;Zuo Chong;Zhao Rui;Sui Xin;Ye Linzhi;Yu Yuanyuan;Chen Jingnan;Wu Xiangwei;Zhang Wenhao;Deng Haiteng;Shi Jing;Pan Man;Li Yi-Ming;Liu Lei
基于三唑的去泛素化酶（DUB）抗性泛素（Ub）探针最近成为蛋白质组学研究中发现Ub链特异性相互作用物的有效工具，但其结构多样性有限。报道了一个新的DUB抗性Ub探针家族，该家族基于异肽-N-乙基化的二聚或聚合Ub链，其可以通过重组Ub前体的一锅、泛素活化酶（E1）催化的缩合反应有效地制备，以得到多毫克规模的各种同型甚至分支Ub探针。使用无标记定量（LFQ）MS的蛋白质组学研究表明，异肽-N-乙基化Ub探针可以在Ub相互作用组研究中补充基于三唑的探针。我们的研究强调了现代蛋白质合成化学在开发蛋白质组学研究所需的结构和新的工具分子家族方面的效用。
Triazole‐based deubiquitylase (DUB)‐resistant ubiquitin (Ub) probes have recently emerged as effective tools for the discovery of Ub chain‐specific interactors in proteomic studies, but their structural diversity is limited. A new family of DUB‐resistant Ub probes is reported based on isopeptide‐N‐ethylated dimeric or polymeric Ub chains, which can be efficiently prepared by a one‐pot, ubiquitin‐activating enzyme (E1)‐catalyzed condensation reaction of recombinant Ub precursors to give various homotypic and even branched Ub probes at multi‐milligram scale. Proteomic studies using label‐free quantitative (LFQ) MS indicated that the isopeptide‐N‐ethylated Ub probes may complement the triazole‐based probes in the study of Ub interactome. Our study highlights the utility of modern protein synthetic chemistry to develop structurally and new families of tool molecules needed for proteomic studies.