Li Chuntong, Wang Tian, Liang Lujun, Chu Guochao, Zhang Jiachen, He Wei, Liu Lei, Li Jinghong
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.
Center for BioAnalytical Chemistry, Hefei National Laboratory of Physical Science at Microscale, University of Science and Technology of China, Hefei, 230026 China.
Sci China Chem. 2023;66(3):837-844. doi: 10.1007/s11426-022-1455-x. Epub 2023 Jan 11.
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. cross-linking 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.
Supplementary material is available for this article at 10.1007/s11426-022-1455-x and is accessible for authorized users.
ISG15是一种类泛素（Ubl）蛋白，通过ISG15缀合酶附着于底物蛋白，其失调与多种疾病过程有关，但对这些酶的探究仍有待完成。在此，我们描述了一种通过蛋白质半合成开发的基于活性的新型探针ISG15-Dha（脱氢丙氨酸）。交联和细胞裂解物蛋白质组分析实验表明，该探针可以依次捕获ISG15缀合酶，包括E1酶UBA7、E2酶UBE2L6、E3酶HERC5、先前已知的ISG15去缀合酶（USP18），以及我们另外确认具有去ISGylation活性的一些其他酶（USP5和USP14）。总体而言，ISG15-Dha提供了一种新工具，可同时捕获ISG15缀合酶和去缀合酶用于生化或药理学研究。
本文的补充材料可在10.1007/s11426-022-1455-x获取，授权用户可访问。
