Photocaging of Activity‐Based Ubiquitin Probes via a C‐Terminal Backbone Modification Strategy. (19th May 2022)

• Record Type: Journal Article
• Authors: Wang, Yu; Chen, Jingnan; Hua, Xiao; Meng, Xianbin; Cai, Hongyi; Wang, Rongtian; Shi, Jing; Deng, Haiteng; Liu, Lei; Li, Yi‐Ming
• Is Part Of: Angewandte Chemie. Volume 134:Number 28(2022)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2022-05-19
• Subjects: Activity-Based Probes -- Chemical Protein Synthesis -- Deubiquitinases -- Photocaging -- Proteomics; Chemistry -- Periodicals; 540
• DOI: 10.1002/ange.202203792
• Languages: English
• ISSNs: 0044-8249

Abstract

Abstract: Photocaged, activity‐based ubiquitin (Ub) probes (Ub‐ABPs) have been developed for the time‐resolved probing of deubiquitinating enzyme (DUB) activities, but many Ub‐ABPs are still challenging to photocage because their warheads (e.g. propargylamide (PA) or dehydroalanine (Dha)) are difficult to temporally block and activate. Here, we describe a new C‐terminal backbone modification strategy for the construction of photocaged Ub‐ABPs in which a light‐sensitive group is placed at the backbone amide bond of the Ub Gly75. This strategy enabled the facile generation of cell‐permeable photocaged Ub‐PA and Dha probes that could be activated to capture DUBs after photo‐irradiation, and were used to profile DUBs in cells under specially designed conditions (e.g. in cells experiencing oxidative stress) or DUBs with isopeptide linkage selectivity. This backbone modification strategy is anticipated to provide a general solution for the development of photocaged Ub ABPs bearing any warheads for DUB profiling. Abstract : Photocaged cell‐permeable Ub‐PA and diUb‐Dha probes that can be activated on demand to label DUBs in vivo have been developed using a C‐terminal backbone modification strategy. These new Ub‐ABPs are temporally blocked and activated through installing a light‐sensitive group (e.g. nitrobenzyl) at the backbone amide bond of the Ub Gly75 residue.