Simultaneous capture of ISG15 conjugating and deconjugating enzymes using a semi-synthetic ISG15-Dha probe  

作　　者：Chuntong Li Tian Wang Lujun Liang Guochao Chu Jiachen Zhang Wei He Lei Liu Jinghong Li 

机构地区：[1]Department of Chemistry,Key Laboratory of Bioorganic Phosphorus Chemistry&Chemical Biology,Center for Synthetic and Systems Biology,State Key Laboratory of Chemical Oncogenomics(Shenzhen),Tsinghua University,Beijing 100084,China [2]School of Pharmaceutical Sciences,Tsinghua University,Beijing 100084,China [3]Center for BioAnalytical Chemistry,Hefei National Laboratory of Physical Science at Microscale,University of Science and Technology of China,Hefei 230026,China

出　　处：《Science China Chemistry》2023年第3期837-844,共8页中国科学（化学英文版）

基　　金：supported by the National Key Research&Development Program of China(2021YFA1200104,2022YFC3401500);the National Natural Science Foundation of China(21621003,22137005,21971133,22027807,22034004,92253302,22227810);the Tsinghua University Spring Breeze Fund(2020Z99CFY043,2021Z99CFZ002)。

摘　　要：ISG15 is a ubiquitin-like(Ubl) protein attached to substrate proteins by ISG15 conjugating enzymes whose dysregulation is implicated in a multitude of disease processes, but the probing of these enzymes remains to be accomplished. Here, we describe the development of a new activity-based probe ISG15-Dha(dehydroalanine) through protein semi-synthesis. In vitro crosslinking and cell lysate proteomic profiling experiments showed that this probe can sequentially capture ISG15 conjugating enzymes including E1 enzyme UBA7, E2 enzyme UBE2L6, E3 enzyme HERC5, the previously known ISG15 deconjugating enzyme(USP18), as well as some other enzymes(USP5 and USP14) which we additionally confirmed to impart deISGylation activity. Collectively, ISG15-Dha provides a new tool that can simultaneously capture ISG15 conjugating and deconjugating enzymes for biochemical or pharmacological studies.

关 键 词：ISG15-Dha probe SEMI-SYNTHESIS activity based probe PROTEOMICS ISGylation 

分 类 号：R37[医药卫生—病原生物学]