Lysine glutarylation is a protein posttranslational modification regulated by SIRT5.
We report the identification and characterization of a five-carbon protein posttranslational modification (PTM) called lysine glutarylation (Kglu). This protein modification was detected by immunoblot and mass spectrometry (MS), and then comprehensively validated by chemical and biochemical methods. We demonstrated that the previously annotated deacetylase, sirtuin 5 (SIRT5), is a lysine deglutarylase. Proteome-wide analysis identified 683 Kglu sites in 191 proteins and showed that Kglu is highly enriched on metabolic enzymes and mitochondrial proteins. We validated carbamoyl phosphate synthase 1 (CPS1), the rate-limiting enzyme in urea cycle, as a glutarylated protein and demonstrated that CPS1 is targeted by SIRT5 for deglutarylation. We further showed that glutarylation suppresses CPS1 enzymatic activity in cell lines, mice, and a model of glutaric acidemia type I disease, the last of which has elevated glutaric acid and glutaryl-CoA. This study expands the landscape of lysine acyl modifications and increases our understanding of the deacylase SIRT5.
The Chemical Proteomics Center and State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, People's Republic of China.
Minjia
M
Tan
Ben May Department for Cancer Research, The University of Chicago, Chicago, IL 60637, USA.
Chao
C
Peng
Sarah W. Stedman Nutrition and Metabolism Center and Department of Medicine, Duke University Medical Center, Durham, NC 27704, USA.
Kristin A
KA
Anderson
Sarah W. Stedman Nutrition and Metabolism Center and Department of Medicine, Duke University Medical Center, Durham, NC 27704, USA.
Peter
P
Chhoy
Ben May Department for Cancer Research, The University of Chicago, Chicago, IL 60637, USA.
Zhongyu
Z
Xie
Ben May Department for Cancer Research, The University of Chicago, Chicago, IL 60637, USA.
Lunzhi
L
Dai
Department of Pathology and Institute of Gerontology, University of Michigan, Ann Arbor, MI 48109, USA.
Jeongsoon
J
Park
Ben May Department for Cancer Research, The University of Chicago, Chicago, IL 60637, USA.
Yue
Y
Chen
Ben May Department for Cancer Research, The University of Chicago, Chicago, IL 60637, USA.
He
H
Huang
The Chemical Proteomics Center and State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, People's Republic of China.
Yi
Y
Zhang
Department of Molecular and Integrative Physiology and Geriatrics Center, University of Michigan, Ann Arbor, MI 48109, USA.
Jennifer
J
Ro
Sarah W. Stedman Nutrition and Metabolism Center and Department of Medicine, Duke University Medical Center, Durham, NC 27704, USA.
Gregory R
GR
Wagner
Sarah W. Stedman Nutrition and Metabolism Center and Department of Medicine, Duke University Medical Center, Durham, NC 27704, USA.
Michelle F
MF
Green
Department of Chemistry, Technical University of Denmark, Kemitorvet 207, DK-2800 Kongens Lyngby, Denmark.
Andreas S
AS
Madsen
Department of Biochemistry, Children's Hospital, University Medical Center Hamburg-Eppendorf, 20246 Hamburg, Germany.
Jessica
J
Schmiesing
Sarah W. Stedman Nutrition and Metabolism Center and Department of Medicine, Duke University Medical Center, Durham, NC 27704, USA.
Brett S
BS
Peterson
Ben May Department for Cancer Research, The University of Chicago, Chicago, IL 60637, USA.
Guofeng
G
Xu
Sarah W. Stedman Nutrition and Metabolism Center and Department of Medicine, Duke University Medical Center, Durham, NC 27704, USA.
Olga R
OR
Ilkayeva
Sarah W. Stedman Nutrition and Metabolism Center and Department of Medicine, Duke University Medical Center, Durham, NC 27704, USA.
Michael J
MJ
Muehlbauer
Department of Biochemistry, Children's Hospital, University Medical Center Hamburg-Eppendorf, 20246 Hamburg, Germany.
Thomas
T
Braulke
Department of Biochemistry, Children's Hospital, University Medical Center Hamburg-Eppendorf, 20246 Hamburg, Germany.
Chris
C
Mühlhausen
Computational Chemistry and Biology Core Facility, Skaggs School of Pharmacy and Pharmaceutical Sciences, University of Colorado Anschutz Medical Campus, Aurora, CO 80045, USA.
Donald S
DS
Backos
Department of Chemistry, Technical University of Denmark, Kemitorvet 207, DK-2800 Kongens Lyngby, Denmark.
Christian A
CA
Olsen
National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892, USA.
Peter J
PJ
McGuire
Department of Molecular and Integrative Physiology and Geriatrics Center, University of Michigan, Ann Arbor, MI 48109, USA.
Scott D
SD
Pletcher
Department of Pathology and Institute of Gerontology, University of Michigan, Ann Arbor, MI 48109, USA.
David B
DB
Lombard
Sarah W. Stedman Nutrition and Metabolism Center and Department of Medicine, Duke University Medical Center, Durham, NC 27704, USA. Electronic address: matthew.hirschey@duke.edu.
Matthew D
MD
Hirschey
The Chemical Proteomics Center and State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, People's Republic of China; Ben May Department for Cancer Research, The University of Chicago, Chicago, IL 60637, USA. Electronic address: yingming.zhao@uchicago.edu.
Yingming
Y
Zhao
D000214:Acyl Coenzyme A / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D002222:Carbamoyl-Phosphate Synthase (Ammonia) / Q000378:metabolism*; D015151:Immunoblotting; D008239:Lysine / Q000737:chemistry / Q000378:metabolism*; D013058:Mass Spectrometry; D051379:Mice; D008954:Models, Biological; D015394:Molecular Structure; D011499:Protein Processing, Post-Translational / Q000502:physiology*; D040901:Proteomics; D037761:Sirtuins / Q000378:metabolism*
D000214:Acyl Coenzyme A; C542554:SIRT5 protein, mouse; C015901:glutaryl-coenzyme A; D037761:Sirtuins; D002222:Carbamoyl-Phosphate Synthase (Ammonia); D008239:Lysine