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. 2023 Jul 3;29(37):e202300414.
doi: 10.1002/chem.202300414. Epub 2023 May 10.

Preparation of UFM1-Derived Probes through Highly Optimized Total Chemical Synthesis

Cong Chen  1 , Chengpiao Lu  1 , Fang Li  1 , Haibo Wang  1 , Shuai Peng  1 , Guoqiang Xu  1 , Hongrui Xu  2 , Jia-Bin Li  1
Affiliations

Preparation of UFM1-Derived Probes through Highly Optimized Total Chemical Synthesis

Cong Chen et al. Chemistry. .

Abstract

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.

Keywords: UFM1; UFM1-derived probe; chemical protein synthesis; fluorescent probe; total chemical synthesis.

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