Genetic code expansion reveals site-specific lactylation in living cells reshapes protein functions

IF 14.7 1区 综合性期刊 Q1 MULTIDISCIPLINARY SCIENCES
Chang Shao, Shuo Tang, Siqin Yu, Chenguang Liu, Yueyang Zhang, Tianyan Wan, Zimeng He, Qi Yuan, Shihan Wu, Hanqing Zhang, Ning Wan, Mengru Zhan, Ren Xiang Tan, Haiping Hao, Hui Ye, Nanxi Wang
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Abstract

Protein lactylation is an emerging field. To advance the exploration of its biological functions, here we develop a comprehensive workflow that integrates proteomics to identify lactylated sites, genetic code expansion (GCE) for the expression of site-specifically lactylated proteins in living cells, and an integrated functional analysis (IFA) platform to evaluate their biological effects. Using a combined wet-and-dry-lab proteomics strategy, we identify a conserved lactylation at ALDOA-K147, which we hypothesize plays a significant biological role. Expression of this site-specifically lactylated ALDOA in mammalian cells reveals that this modification not only inhibits enzymatic activity but also induces gain-of-function effects. These effects reshaped ALDOA functionality by enhancing protein stability, promoting nuclear translocation, regulating adhesion-related gene expression, altering cell morphology and modulating ALDOA-interacting proteins. Our findings highlight the utility of the GCE-based workflow in establishing causal relationships between specific lactylation events and both target-specific and cell-wide changes, advancing our understanding of protein lactylation’s functional impact.

Abstract Image

遗传密码扩展揭示了活细胞中位点特异性乳酸化重塑蛋白质功能
蛋白质乳酸化是一个新兴的领域。为了推进其生物学功能的探索,我们开发了一个综合的工作流程,包括蛋白质组学来识别乳酸化位点,遗传密码扩增(GCE)来表达活细胞中特定位点的乳酸化蛋白,以及综合功能分析(IFA)平台来评估其生物学效应。使用干湿联合实验室蛋白质组学策略,我们确定了ALDOA-K147的保守乳酸化,我们假设它起着重要的生物学作用。哺乳动物细胞中这种位点特异性乳酸化ALDOA的表达表明,这种修饰不仅抑制酶活性,而且诱导功能获得效应。这些影响通过增强蛋白质稳定性、促进核易位、调节粘附相关基因表达、改变细胞形态和调节ALDOA相互作用蛋白来重塑ALDOA功能。我们的研究结果强调了基于gce的工作流程在建立特异性乳酸化事件与靶标特异性和细胞范围变化之间的因果关系方面的效用,促进了我们对蛋白质乳酸化功能影响的理解。
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来源期刊
Nature Communications
Nature Communications Biological Science Disciplines-
CiteScore
24.90
自引率
2.40%
发文量
6928
审稿时长
3.7 months
期刊介绍: Nature Communications, an open-access journal, publishes high-quality research spanning all areas of the natural sciences. Papers featured in the journal showcase significant advances relevant to specialists in each respective field. With a 2-year impact factor of 16.6 (2022) and a median time of 8 days from submission to the first editorial decision, Nature Communications is committed to rapid dissemination of research findings. As a multidisciplinary journal, it welcomes contributions from biological, health, physical, chemical, Earth, social, mathematical, applied, and engineering sciences, aiming to highlight important breakthroughs within each domain.
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