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Efficient Chemical Synthesis of Multi-Monoubiquitylated and Diubiquitylated Histones by the α‑Halogen Ketone-Mediated Strategy

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posted on 2024-07-02, 19:44 authored by Shuai Peng, Xin Liu, Chengpiao Lu, Haibo Wang, Xiaotong Liu, Qingyue Gong, Huizhong Tao, Hongrui Xu, Changlin Tian, Guoqiang Xu, Jia-Bin Li
The chemical synthesis of homogeneously ubiquitylated histones is a powerful approach to decipher histone ubiquitylation-dependent epigenetic regulation. Among the various methods, α-halogen ketone-mediated conjugation chemistry has recently been an attractive strategy to generate single-monoubiquitylated histones for biochemical and structural studies. Herein, we report the use of this strategy to prepare not only dual- and even triple-monoubiquitylated histones but also diubiquitin-modified histones. We were surprised to find that the synthetic efficiencies of multi-monoubiquitylated histones were comparable to those of single-monoubiquitylated ones, suggesting that this strategy is highly tolerant to the number of ubiquitin monomers installed onto histones. The facile generation of a series of single-, dual-, and triple-monoubiquitylated H3 proteins enabled us to evaluate the influence of ubiquitylation patterns on the binding of DNA methyltransferase 1 (DNMT1) to nucleosomes. Our study highlights the potential of site-specific conjugation chemistry to generate chemically defined histones for epigenetic studies.

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