Hepatic glutamine synthetase controls N5-methylglutamine in homeostasis and cancer

VH Villar, MF Allega, R Deshmukh, T Ackermann, MA Nakasone, J Vande Voorde, TM Drake, J Oetjen, A Bloom, C Nixon, M Müller, S May, EH Tan, L Vereecke, M Jans, G Blancke, DJ Murphy, DT Huang, DY Lewis, TG BirdOJ Sansom, K Blyth, D Sumpton, S Tardito*

*Corresponding author for this work

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Glutamine synthetase (GS) activity is conserved from prokaryotes to humans, where the ATP-dependent production of glutamine from glutamate and ammonia is essential for neurotransmission and ammonia detoxification. Here, we show that mammalian GS uses glutamate and methylamine to produce a methylated glutamine analog, N5-methylglutamine. Untargeted metabolomics revealed that liver-specific GS deletion and its pharmacological inhibition in mice suppress hepatic and circulating levels of N5-methylglutamine. This alternative activity of GS was confirmed in human recombinant enzyme and cells, where a pathogenic mutation in the active site (R324C) promoted the synthesis of N5-methylglutamine over glutamine. N5-methylglutamine is detected in the circulation, and its levels are sustained by the microbiome, as demonstrated by using germ-free mice. Finally, we show that urine levels of N5-methylglutamine correlate with tumor burden and GS expression in a β-catenin-driven model of liver cancer, highlighting the translational potential of this uncharacterized metabolite.
Original languageEnglish
Pages (from-to)292-304
Number of pages13
JournalNature Chemical Biology
Issue number3
Early online date24 Oct 2022
Publication statusPublished - Mar 2023


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