Site-Specific incorporation of lysine acetyl-methylation into proteins

IF 3.1 4区医学 Q3 CHEMISTRY, MEDICINAL

Medicinal Chemistry Research Pub Date : 2026-03-25 DOI:10.1007/s00044-026-03544-3

Jingxiang Zhao, Xinlong Guo, Xuelian Ren, Jiyang Yu, Xiaohan Song, He Huang

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引用次数: 0

Abstract

Lysine acetyl-methylation (Kam) is a novel protein post-translational modification (PTM) recently identified on histone H4, yet its biological functions on non-histone proteins remain largely unexplored. Here, we engineered an orthogonal Methanosarcina barkeri pyrrolysyl-tRNA synthetase (MbPylRS) variant and tRNA_CUA^Pyl pair for site-specific Kam incorporation into proteins in mammalian cells. Using this system, we identified endogenous Kam sites at K28 of adenylate kinase 2 (AK2) and K207 of pyruvate kinase M2 (PKM2) through proteomic analysis. Functional characterization revealed that Kam at these evolutionarily conserved residues significantly attenuated enzymatic activity. These findings demonstrate that Kam serves as a regulatory mechanism for non-histone proteins, and the ability to generate proteins harboring Kam at defined sites provides a valuable approach for investigating the biological roles of this newly identified PTM.

The alternative text for this image may have been generated using AI.

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赖氨酸乙酰甲基化与蛋白质的位点特异性结合

赖氨酸乙酰甲基化（Kam）是最近在组蛋白H4上发现的一种新的蛋白质翻译后修饰（PTM），但其在非组蛋白上的生物学功能仍未被广泛探索。在这里，我们设计了一个正交的Methanosarcina barkeri pyrlyyl - trna合成酶（MbPylRS）变体和tRNACUAPyl对，用于在哺乳动物细胞中将位点特异性的Kam整合到蛋白质中。利用该系统，我们通过蛋白质组学分析确定了腺苷酸激酶2 （AK2）的K28位点和丙酮酸激酶M2 （PKM2）的K207位点的内源性Kam位点。功能表征表明，Kam at这些进化上保守的残基显著减弱了酶的活性。这些发现表明，Kam是一种非组蛋白的调控机制，而在特定位点产生含有Kam的蛋白质的能力为研究这种新发现的PTM的生物学作用提供了一种有价值的方法。此图像的替代文本可能是使用AI生成的。

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来源期刊

Medicinal Chemistry Research 医学-医药化学

CiteScore

4.70

自引率

3.80%

发文量

162

审稿时长

5.0 months

期刊介绍： Medicinal Chemistry Research (MCRE) publishes papers on a wide range of topics, favoring research with significant, new, and up-to-date information. Although the journal has a demanding peer review process, MCRE still boasts rapid publication, due in part, to the length of the submissions. The journal publishes significant research on various topics, many of which emphasize the structure-activity relationships of molecular biology.