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We report identification and characterization of a new protein post-translational modification (PTM),lysine glutarylation (Kglu).This modification was initially detected by Western blot using an anti-Kglu specific antibody (custom made by PTM Biolabs,Inc.) and mass spectrometry (MS),and comprehensively validated by isotopic labeling,HPLC coelution and MS/MS.We further showed that Kglu was an evolutionarily-conserved PTM that exists in both bacterial and mammalian cells.