Preparation of UFM1-Derived Probes through Highly Optimized Total Chemical Synthesis
Preparation of UFM1-Derived Probes through Highly Optimized Total Chemical Synthesis
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DOI：
10.1002/chem.202300414
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发表时间：
2023-05-10
影响因子：
4.3
通讯作者：
Li，Jia-Bin
中科院分区：
文献类型：
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作者：
Chen，Cong;Lu，Chengpiao;Li，Jia-Bin
ufmyation参与多种细胞过程，并与许多人类疾病有关。对这种修饰的理解依赖于使用定制的UFM1衍生探针对其相关酶进行基于活性的分析。本研究提出了一种高度优化的全化学合成方法，用于生成多种UFM1衍生的探针，包括UFM1‐PA、Biotin‐UFM1‐PA和UFM1‐AMC，其中UFM1 C末端缬氨酸酰肼很容易通过酰肼连接制备，并用作安装酶敏感弹头和荧光报告器的多功能手柄。所得探针在细胞裂解物中对UFM1特异性酶显示出高反应性和选择性。这一策略促进了UFM1衍生工具包的生成和多样性，这些工具包可用于分析UFM1特异性酶，从而对UFM1化的动力学产生深刻的见解。
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.