Genetic code expansion reveals site-specific lactylation in living cells reshapes protein functions
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 liv...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2025-01-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-024-55165-2 |
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| _version_ | 1841544488994996224 |
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| author | 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 |
| author_facet | 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 |
| author_sort | Chang Shao |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-0d867969f4d84c4c80996918f6d4c3bf |
| institution | Kabale University |
| issn | 2041-1723 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj-art-0d867969f4d84c4c80996918f6d4c3bf2025-01-12T12:30:18ZengNature PortfolioNature Communications2041-17232025-01-0116111610.1038/s41467-024-55165-2Genetic code expansion reveals site-specific lactylation in living cells reshapes protein functionsChang Shao0Shuo Tang1Siqin Yu2Chenguang Liu3Yueyang Zhang4Tianyan Wan5Zimeng He6Qi Yuan7Shihan Wu8Hanqing Zhang9Ning Wan10Mengru Zhan11Ren Xiang Tan12Haiping Hao13Hui Ye14Nanxi Wang15Jiangsu Provincial Key Laboratory of Drug Metabolism and Pharmacokinetics, State Key Laboratory of Natural Medicines, China Pharmaceutical University, Tongjiaxiang No. 24School of Pharmacy, Nanjing University of Chinese Medicine, Xianlindadao No. 138Jiangsu Provincial Key Laboratory of Drug Metabolism and Pharmacokinetics, State Key Laboratory of Natural Medicines, China Pharmaceutical University, Tongjiaxiang No. 24Jiangsu Provincial Key Laboratory of Drug Metabolism and Pharmacokinetics, State Key Laboratory of Natural Medicines, China Pharmaceutical University, Tongjiaxiang No. 24Jiangsu Provincial Key Laboratory of Drug Metabolism and Pharmacokinetics, State Key Laboratory of Natural Medicines, China Pharmaceutical University, Tongjiaxiang No. 24School of Pharmacy, Nanjing University of Chinese Medicine, Xianlindadao No. 138Jiangsu Provincial Key Laboratory of Drug Metabolism and Pharmacokinetics, State Key Laboratory of Natural Medicines, China Pharmaceutical University, Tongjiaxiang No. 24Jiangsu Provincial Key Laboratory of Drug Metabolism and Pharmacokinetics, State Key Laboratory of Natural Medicines, China Pharmaceutical University, Tongjiaxiang No. 24School of Pharmacy, Nanjing University of Chinese Medicine, Xianlindadao No. 138Jiangsu Provincial Key Laboratory of Drug Metabolism and Pharmacokinetics, State Key Laboratory of Natural Medicines, China Pharmaceutical University, Tongjiaxiang No. 24Jiangsu Provincial Key Laboratory of Drug Metabolism and Pharmacokinetics, State Key Laboratory of Natural Medicines, China Pharmaceutical University, Tongjiaxiang No. 24School of Pharmacy, Nanjing University of Chinese Medicine, Xianlindadao No. 138School of Pharmacy, Nanjing University of Chinese Medicine, Xianlindadao No. 138Jiangsu Provincial Key Laboratory of Drug Metabolism and Pharmacokinetics, State Key Laboratory of Natural Medicines, China Pharmaceutical University, Tongjiaxiang No. 24Jiangsu Provincial Key Laboratory of Drug Metabolism and Pharmacokinetics, State Key Laboratory of Natural Medicines, China Pharmaceutical University, Tongjiaxiang No. 24School of Pharmacy, Nanjing University of Chinese Medicine, Xianlindadao No. 138Abstract 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.https://doi.org/10.1038/s41467-024-55165-2 |
| spellingShingle | 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 Genetic code expansion reveals site-specific lactylation in living cells reshapes protein functions Nature Communications |
| title | Genetic code expansion reveals site-specific lactylation in living cells reshapes protein functions |
| title_full | Genetic code expansion reveals site-specific lactylation in living cells reshapes protein functions |
| title_fullStr | Genetic code expansion reveals site-specific lactylation in living cells reshapes protein functions |
| title_full_unstemmed | Genetic code expansion reveals site-specific lactylation in living cells reshapes protein functions |
| title_short | Genetic code expansion reveals site-specific lactylation in living cells reshapes protein functions |
| title_sort | genetic code expansion reveals site specific lactylation in living cells reshapes protein functions |
| url | https://doi.org/10.1038/s41467-024-55165-2 |
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