Department of Chemistry, Key Laboratory of Bioorganic Phosphorus Chemistry & Chemical Biology, Tsinghua University, Beijing, 100084, P. R. China.
School of Pharmaceutical Sciences, Tsinghua University, Beijing, 100084, P. R. China.
Angew Chem Int Ed Engl. 2022 Oct 4;61(40):e202206205. doi: 10.1002/anie.202206205. Epub 2022 Aug 29.
Ubiquitin (Ub)-like protein ISG15 (interferon-stimulated gene 15) regulates innate immunity and links with the evasion of host response by viruses such as SARS-CoV-2. Dissecting ISGylation pathways recently received increasing attention which can inform related disease interventions, but such studies necessitate the preparation and development of various ISG15 protein tools. Here, we find that the leader protease (Lb ) encoded by foot-and-mouth disease virus can promote ligation reactions between recombinant ISG15 and synthetic glycyl compounds, generating protein tools such as ISG15-propargylamide and ISG15-rhodamine110, which are needed for cellular proteomic studies of deISGylases, and the screening and evaluation of inhibitors against SARS-CoV-2 papain-like protease (PLpro). Furthermore, this strategy can be also used to load ISG15 onto the lysine of a synthetic peptide through an isopeptide bond, and prepare Ub and NEDD8 (ubiquitin-like protein Nedd8) protein tools.
泛素样蛋白 ISG15（干扰素刺激基因 15）调节先天免疫，并与 SARS-CoV-2 等病毒逃避宿主反应有关。最近，对 ISG 化途径的分析受到越来越多的关注，这可以为相关疾病的干预提供信息，但此类研究需要制备和开发各种 ISG15 蛋白工具。在这里，我们发现口蹄疫病毒编码的 3C 蛋白酶可以促进重组 ISG15 和合成甘氨酰化合物之间的连接反应，生成 ISG15-炔丙酰胺和 ISG15-罗丹明 110 等蛋白工具，这些工具是细胞中去 ISG 酶研究所必需的，也是筛选和评估针对 SARS-CoV-2 木瓜蛋白酶样蛋白酶（PLpro）抑制剂所必需的。此外，该策略还可以用于通过异肽键将 ISG15 加载到合成肽的赖氨酸上，并制备 Ub 和 NEDD8（泛素样蛋白 Nedd8）蛋白工具。
