Department of Biochemistry, University of Texas Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, Texas 75390-9038, United States.
Departments of Biophysics and Microbiology, University of Texas Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, Texas 75390-9038, United States.
ACS Chem Biol. 2022 Apr 15;17(4):810-815. doi: 10.1021/acschembio.2c00091. Epub 2022 Mar 21.
Recently developed chemical and enzyme-based technologies to install serine ADP-ribosylation onto synthetic peptides have enabled new approaches to study poly(ADP-ribose) polymerase (PARP) biology. Here, we establish a generalizable strategy to prepare ADP-ribosylated peptides that are compatible with N-terminal, C-terminal, and sequential protein ligation reactions. Two unique protein-assembly routes are employed to generate full-length linker histone constructs that are homogeneously ADP-ribosylated at known DNA damage-dependent modification sites. We found that serine mono-ADP-ribosylation is sufficient to alleviate linker histone-dependent chromatin compaction and that this effect is amplified by ADP-ribose chain elongation. Our work will greatly expand the scope of ADP-ribose-modified proteins that can be constructed via semisynthesis, which is rapidly emerging as a robust approach to elucidate the direct effects that site-specific serine mono- and poly-ADP-ribosylation have on protein function.
最近开发的化学和酶基技术可将丝氨酸 ADP-核糖基化安装到合成肽上，为研究聚（ADP-核糖）聚合酶（PARP）生物学开辟了新途径。在这里，我们建立了一种可普遍应用的策略来制备 ADP-核糖基化肽，这些肽与 N 端、C 端和顺序蛋白连接反应兼容。采用两种独特的蛋白质组装途径来生成全长连接组蛋白构建体，这些构建体在已知的 DNA 损伤依赖性修饰位点处均一 ADP-核糖基化。我们发现丝氨酸单 ADP-核糖基化足以减轻连接组蛋白依赖性染色质紧缩，并且这种效应通过 ADP-核糖链延伸而放大。我们的工作将大大扩展可以通过半合成构建的 ADP-核糖修饰蛋白的范围，这是一种迅速发展的强大方法，可以阐明特定丝氨酸单和多 ADP-核糖基化对蛋白质功能的直接影响。
