Charting unknown metabolic reactions by mass spectrometry-resolved stable-isotope tracing metabolomics

Abstract Metabolic reactions play important roles in organisms such as providing energy, transmitting signals, and synthesizing biomacromolecules. Charting unknown metabolic reactions in cells is hindered by limited technologies, restricting the holistic understanding of cellular metabolism. Using m...

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Main Authors: Yang Gao, Mingdu Luo, Hongmiao Wang, Zhiwei Zhou, Yandong Yin, Ruohong Wang, Beizi Xing, Xiaohua Yang, Yuping Cai, Zheng-Jiang Zhu
Format: Article
Language:English
Published: Nature Portfolio 2025-05-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-025-60258-7
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author Yang Gao
Mingdu Luo
Hongmiao Wang
Zhiwei Zhou
Yandong Yin
Ruohong Wang
Beizi Xing
Xiaohua Yang
Yuping Cai
Zheng-Jiang Zhu
author_facet Yang Gao
Mingdu Luo
Hongmiao Wang
Zhiwei Zhou
Yandong Yin
Ruohong Wang
Beizi Xing
Xiaohua Yang
Yuping Cai
Zheng-Jiang Zhu
author_sort Yang Gao
collection DOAJ
description Abstract Metabolic reactions play important roles in organisms such as providing energy, transmitting signals, and synthesizing biomacromolecules. Charting unknown metabolic reactions in cells is hindered by limited technologies, restricting the holistic understanding of cellular metabolism. Using mass spectrometry-resolved stable-isotope tracing metabolomics, we develop an isotopologue similarity networking strategy, namely IsoNet, to effectively deduce previously unknown metabolic reactions. The strategy uncovers ~300 previously unknown metabolic reactions in living cells and mice. Specifically, we elaborately chart the metabolic reaction network related to glutathione, unveiling three previously unreported reactions nestled within glutathione metabolism. Among these, a transsulfuration reaction, synthesizing γ-glutamyl-seryl-glycine directly from glutathione, underscores the role of glutathione as a sulfur donor. Functional metabolomics studies systematically characterize biochemical effects of previously unknown reactions in glutathione metabolism, showcasing their diverse functions in regulating cellular metabolism. Overall, these newly uncovered metabolic reactions fill gaps in the metabolic network maps, facilitating exploration of uncharted territories in cellular biochemistry.
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institution DOAJ
issn 2041-1723
language English
publishDate 2025-05-01
publisher Nature Portfolio
record_format Article
series Nature Communications
spelling doaj-art-019a9fdcb4bb4f6792ea13f5ff6cea462025-08-20T03:16:41ZengNature PortfolioNature Communications2041-17232025-05-0116111610.1038/s41467-025-60258-7Charting unknown metabolic reactions by mass spectrometry-resolved stable-isotope tracing metabolomicsYang Gao0Mingdu Luo1Hongmiao Wang2Zhiwei Zhou3Yandong Yin4Ruohong Wang5Beizi Xing6Xiaohua Yang7Yuping Cai8Zheng-Jiang Zhu9Interdisciplinary Research Center on Biology and Chemistry, Shanghai Institute of Organic Chemistry, Chinese Academy of SciencesInterdisciplinary Research Center on Biology and Chemistry, Shanghai Institute of Organic Chemistry, Chinese Academy of SciencesInterdisciplinary Research Center on Biology and Chemistry, Shanghai Institute of Organic Chemistry, Chinese Academy of SciencesInterdisciplinary Research Center on Biology and Chemistry, Shanghai Institute of Organic Chemistry, Chinese Academy of SciencesInterdisciplinary Research Center on Biology and Chemistry, Shanghai Institute of Organic Chemistry, Chinese Academy of SciencesInterdisciplinary Research Center on Biology and Chemistry, Shanghai Institute of Organic Chemistry, Chinese Academy of SciencesInterdisciplinary Research Center on Biology and Chemistry, Shanghai Institute of Organic Chemistry, Chinese Academy of SciencesInterdisciplinary Research Center on Biology and Chemistry, Shanghai Institute of Organic Chemistry, Chinese Academy of SciencesInterdisciplinary Research Center on Biology and Chemistry, Shanghai Institute of Organic Chemistry, Chinese Academy of SciencesInterdisciplinary Research Center on Biology and Chemistry, Shanghai Institute of Organic Chemistry, Chinese Academy of SciencesAbstract Metabolic reactions play important roles in organisms such as providing energy, transmitting signals, and synthesizing biomacromolecules. Charting unknown metabolic reactions in cells is hindered by limited technologies, restricting the holistic understanding of cellular metabolism. Using mass spectrometry-resolved stable-isotope tracing metabolomics, we develop an isotopologue similarity networking strategy, namely IsoNet, to effectively deduce previously unknown metabolic reactions. The strategy uncovers ~300 previously unknown metabolic reactions in living cells and mice. Specifically, we elaborately chart the metabolic reaction network related to glutathione, unveiling three previously unreported reactions nestled within glutathione metabolism. Among these, a transsulfuration reaction, synthesizing γ-glutamyl-seryl-glycine directly from glutathione, underscores the role of glutathione as a sulfur donor. Functional metabolomics studies systematically characterize biochemical effects of previously unknown reactions in glutathione metabolism, showcasing their diverse functions in regulating cellular metabolism. Overall, these newly uncovered metabolic reactions fill gaps in the metabolic network maps, facilitating exploration of uncharted territories in cellular biochemistry.https://doi.org/10.1038/s41467-025-60258-7
spellingShingle Yang Gao
Mingdu Luo
Hongmiao Wang
Zhiwei Zhou
Yandong Yin
Ruohong Wang
Beizi Xing
Xiaohua Yang
Yuping Cai
Zheng-Jiang Zhu
Charting unknown metabolic reactions by mass spectrometry-resolved stable-isotope tracing metabolomics
Nature Communications
title Charting unknown metabolic reactions by mass spectrometry-resolved stable-isotope tracing metabolomics
title_full Charting unknown metabolic reactions by mass spectrometry-resolved stable-isotope tracing metabolomics
title_fullStr Charting unknown metabolic reactions by mass spectrometry-resolved stable-isotope tracing metabolomics
title_full_unstemmed Charting unknown metabolic reactions by mass spectrometry-resolved stable-isotope tracing metabolomics
title_short Charting unknown metabolic reactions by mass spectrometry-resolved stable-isotope tracing metabolomics
title_sort charting unknown metabolic reactions by mass spectrometry resolved stable isotope tracing metabolomics
url https://doi.org/10.1038/s41467-025-60258-7
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