title: SIRT5 Regulates both Cytosolic and Mitochondrial Protein Malonylation with Glycolysis as a Major Target.
abstract: 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.
authors.affiliation: Gladstone Institutes and University of California, San Francisco, San Francisco, CA 94158, USA.
authors.forename: Yuya
authors.initials: Y
authors.lastname: Nishida
authors.affiliation: Buck Institute for Research on Aging, 8001 Redwood Boulevard, Novato, CA 94945, USA.
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Rardin
authors.affiliation: Gladstone Institutes and University of California, San Francisco, San Francisco, CA 94158, USA.
authors.forename: Chris
authors.initials: C
authors.lastname: Carrico
authors.affiliation: Gladstone Institutes and University of California, San Francisco, San Francisco, CA 94158, USA.
authors.forename: Wenjuan
authors.initials: W
authors.lastname: He
authors.affiliation: Buck Institute for Research on Aging, 8001 Redwood Boulevard, Novato, CA 94945, USA.
authors.forename: Alexandria K
authors.initials: AK
authors.lastname: Sahu
authors.affiliation: Gladstone Institutes and University of California, San Francisco, San Francisco, CA 94158, USA.
authors.forename: Philipp
authors.initials: P
authors.lastname: Gut
authors.affiliation: Cell Signaling Technology, Inc, 3 Trask Lane, Danvers, MA 01923, USA.
authors.forename: Rami
authors.initials: R
authors.lastname: Najjar
authors.affiliation: Department of Nutritional Sciences and Toxicology, University of California, Berkeley, Berkeley, CA 94720, USA.
authors.forename: Mark
authors.initials: M
authors.lastname: Fitch
authors.affiliation: Department of Nutritional Sciences and Toxicology, University of California, Berkeley, Berkeley, CA 94720, USA; KineMed, Inc., Emeryville, CA 94608, USA.
authors.forename: Marc
authors.initials: M
authors.lastname: Hellerstein
authors.affiliation: Buck Institute for Research on Aging, 8001 Redwood Boulevard, Novato, CA 94945, USA. Electronic address: bgibson@buckinstitute.org.
authors.forename: Bradford W
authors.initials: BW
authors.lastname: Gibson
authors.affiliation: Gladstone Institutes and University of California, San Francisco, San Francisco, CA 94158, USA. Electronic address: everdin@gladstone.ucsf.edu.
authors.forename: Eric
authors.initials: E
authors.lastname: Verdin
mesh_terms: D000215:Acylation; D019943:Amino Acid Substitution; D000818:Animals; D020134:Catalytic Domain; D003600:Cytosol / Q000378:metabolism; D055785:Gene Knockdown Techniques; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D006019:Glycolysis; D057809:HEK293 Cells; D006801:Humans; D008099:Liver / Q000378:metabolism; D008314:Malonates / Q000378:metabolism; D053858:Metabolic Networks and Pathways; D051379:Mice; D018345:Mice, Knockout; D024101:Mitochondrial Proteins / Q000378:metabolism; D018716:Molecular Mimicry; D037761:Sirtuins / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*
chemical_list: D008314:Malonates; D024101:Mitochondrial Proteins; C542554:SIRT5 protein, mouse; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D037761:Sirtuins