SIRT5 Regulates both Cytosolic and Mitochondrial Protein Malonylation with Glycolysis as a Major Target

Mol Cell. 2015 Jul 16;59(2):321-32. doi: 10.1016/j.molcel.2015.05.022. Epub 2015 Jun 11.


Protein acylation links energetic substrate flux with cellular adaptive responses. SIRT5 is a NAD(+)-dependent lysine deacylase and removes both succinyl and malonyl groups. Using affinity enrichment and label free quantitative proteomics, we characterized the SIRT5-regulated lysine malonylome in wild-type (WT) and Sirt5(-/-) mice. 1,137 malonyllysine sites were identified across 430 proteins, with 183 sites (from 120 proteins) significantly increased in Sirt5(-/-) animals. Pathway analysis identified glycolysis as the top SIRT5-regulated pathway. Importantly, glycolytic flux was diminished in primary hepatocytes from Sirt5(-/-) compared to WT mice. Substitution of malonylated lysine residue 184 in glyceraldehyde 3-phosphate dehydrogenase with glutamic acid, a malonyllysine mimic, suppressed its enzymatic activity. Comparison with our previous reports on acylation reveals that malonylation targets a different set of proteins than acetylation and succinylation. These data demonstrate that SIRT5 is a global regulator of lysine malonylation and provide a mechanism for regulation of energetic flux through glycolysis.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acylation
  • Amino Acid Substitution
  • Animals
  • Catalytic Domain
  • Cytosol / metabolism
  • Gene Knockdown Techniques
  • Glyceraldehyde-3-Phosphate Dehydrogenases / chemistry
  • Glyceraldehyde-3-Phosphate Dehydrogenases / genetics
  • Glyceraldehyde-3-Phosphate Dehydrogenases / metabolism
  • Glycolysis
  • HEK293 Cells
  • Humans
  • Liver / metabolism
  • Malonates / metabolism
  • Metabolic Networks and Pathways
  • Mice
  • Mice, Knockout
  • Mitochondrial Proteins / metabolism
  • Molecular Mimicry
  • Sirtuins / deficiency
  • Sirtuins / genetics
  • Sirtuins / metabolism*


  • Malonates
  • Mitochondrial Proteins
  • SIRT5 protein, mouse
  • Glyceraldehyde-3-Phosphate Dehydrogenases
  • Sirtuins