Jiangsu Key Laboratory of Neuropsychiatric Diseases and, College of Pharmaceutical Sciences, Jiangsu Key Laboratory of Preventive and Translational Medicine for Geriatric Diseases, Suzhou Key Laboratory of Drug Research for, Prevention and Treatment of Hyperlipidemic Diseases, Soochow University, Suzhou, 215123, China.
Suzhou Municipal Center for Disease Control and Prevention, Suzhou, Jiangsu, 215004, China.
Chemistry. 2023 Jul 3;29(37):e202300414. doi: 10.1002/chem.202300414. Epub 2023 May 10.
Ufmylation is involved in various cellular processes and associated with many human diseases. The understanding of this modification relies on the use of customized UFM1-derived probes for activity-based profiling of its related enzymes. This study presents a highly optimized total chemical synthesis for the generation of diverse UFM1-derived probes including UFM1-PA, Biotin-UFM1-PA and UFM1-AMC, in which a UFM1 C-terminal valine hydrazide was readily prepared by hydrazide-based ligation and used as a versatile handle for the installation of enzyme-sensitive warheads and fluorescent reporters. The resulting probes display high reactivity and selectivity for UFM1-specific enzymes in cell lysates. This strategy facilitates the generation and diversity of the UFM1-derived toolkit that can be employed to profile UFM1-specific enzymes, thereby shining insights into the dynamics of ufmylation.
泛素样修饰（Ufmylation）参与多种细胞过程，并与许多人类疾病相关。对这种修饰的理解依赖于使用定制的 UFM1 衍生探针来进行其相关酶的基于活性的分析。本研究提出了一种高度优化的总化学合成方法，用于生成多种 UFM1 衍生探针，包括 UFM1-PA、生物素-UFM1-PA 和 UFM1-AMC，其中 UFM1 C 末端缬氨酸酰肼可通过酰肼连接轻易制备，并用作安装酶敏感弹头和荧光报告基团的通用接头。所得探针在细胞裂解物中对 UFM1 特异性酶具有高反应性和选择性。该策略促进了 UFM1 衍生工具包的生成和多样性，可用于分析 UFM1 特异性酶，从而深入了解泛素样修饰的动态。
