Chemoenzymatic and Synthetic Approaches To Investigate Aspartate- and Glutamate-ADP-Ribosylation.
Chemoenzymatic and Synthetic Approaches To Investigate Aspartate- and Glutamate-ADP-Ribosylation.
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研究天冬氨酸和谷氨酸-ADP-核糖基化的化学酶法和合成方法。
DOI：
10.1021/jacs.3c03771
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发表时间：
2023
影响因子：
15
通讯作者：
Tashiro K
中科院分区：
文献类型：
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作者：
Tashiro K
我们在这里报告化学酶法和全合成方法，用 ADP-核糖在肽的特定位点修饰天冬氨酸和谷氨酸侧链。天冬氨酸和谷氨酸 ADP-核糖基化肽的结构分析揭示了侧链键从异头碳到 2"-或 3"-ADP-核糖羟基部分的近定量迁移。我们发现这种连接迁移模式是天冬氨酸和谷氨酸 ADP-核糖基化所独有的，并提出观察到的异构体分布谱存在于生化和细胞环境中。在定义了天冬氨酸和谷氨酸 ADP-核糖基化的不同稳定性特性后，我们设计了在特定谷氨酸位点安装同质 ADP-核糖链并将谷氨酸修饰的肽组装成全长蛋白质的方法。通过实施这些技术，我们表明组蛋白 H2B E2 三 ADP 核糖基化能够刺激染色质重塑器 ALC1，其效率与组蛋白丝氨酸 ADP 核糖基化相似。我们的工作揭示了天冬氨酸和谷氨酸 ADP-核糖基化的基本原理，并提供了新的策略来探究这种广泛的蛋白质修饰的生化后果。
We report here chemoenzymatic and fully synthetic methodologies to modify aspartate and glutamate side chains with ADP-ribose at specific sites on peptides. Structural analysis of aspartate and glutamate ADP-ribosylated peptides reveals near-quantitative migration of the side chain linkage from the anomeric carbon to the 2″- or 3″-ADP-ribose hydroxyl moieties. We find that this linkage migration pattern is unique to aspartate and glutamate ADP-ribosylation and propose that the observed isomer distribution profile is present in biochemical and cellular environments. After defining distinct stability properties of aspartate and glutamate ADP-ribosylation, we devise methods to install homogenous ADP-ribose chains at specific glutamate sites and assemble glutamate-modified peptides into full-length proteins. By implementing these technologies, we show that histone H2B E2 tri-ADP-ribosylation is able to stimulate the chromatin remodeler ALC1 with similar efficiency to histone serine ADP-ribosylation. Our work reveals fundamental principles of aspartate and glutamate ADP-ribosylation and enables new strategies to interrogate the biochemical consequences of this widespread protein modification.