title: A Class of Reactive Acyl-CoA Species Reveals the Non-enzymatic Origins of Protein Acylation.
abstract: The mechanisms underlying the formation of acyl protein modifications remain poorly understood. By investigating the reactivity of endogenous acyl-CoA metabolites, we found a class of acyl-CoAs that undergo intramolecular catalysis to form reactive intermediates that non-enzymatically modify proteins. Based on this mechanism, we predicted, validated, and characterized a protein modification: 3-hydroxy-3-methylglutaryl(HMG)-lysine. In a model of altered HMG-CoA metabolism, we found evidence of two additional protein modifications: 3-methylglutaconyl(MGc)-lysine and 3-methylglutaryl(MG)-lysine. Using quantitative proteomics, we compared the "acylomes" of two reactive acyl-CoA species, namely HMG-CoA and glutaryl-CoA, which are generated in different pathways. We found proteins that are uniquely modified by each reactive metabolite, as well as common proteins and pathways. We identified the tricarboxylic acid cycle as a pathway commonly regulated by acylation and validated malate dehydrogenase as a key target. These data uncover a fundamental relationship between reactive acyl-CoA species and proteins and define a new regulatory paradigm in metabolism.
authors.affiliation: Sarah W. Stedman Nutrition and Metabolism Center, Duke Molecular Physiology Institute, Duke University Medical Center, Durham, NC 27710, USA; Division of Endocrinology, Metabolism, and Nutrition, Department of Medicine, Duke University Medical Center, Durham, NC 27710, USA.
authors.forename: Gregory R
authors.initials: GR
authors.lastname: Wagner
authors.affiliation: Sarah W. Stedman Nutrition and Metabolism Center, Duke Molecular Physiology Institute, Duke University Medical Center, Durham, NC 27710, USA.
authors.forename: Dhaval P
authors.initials: DP
authors.lastname: Bhatt
authors.affiliation: Sarah W. Stedman Nutrition and Metabolism Center, Duke Molecular Physiology Institute, Duke University Medical Center, Durham, NC 27710, USA.
authors.forename: Thomas M
authors.initials: TM
authors.lastname: O'Connell
authors.affiliation: Department of Pharmacology & Cancer Biology, Duke University Medical Center, Durham, NC 27710, USA; Duke Proteomics and Metabolomics Shared Resource, Duke University Medical Center, Durham, NC 27710, USA.
authors.forename: J Will
authors.initials: JW
authors.lastname: Thompson
authors.affiliation: Duke Proteomics and Metabolomics Shared Resource, Duke University Medical Center, Durham, NC 27710, USA.
authors.forename: Laura G
authors.initials: LG
authors.lastname: Dubois
authors.affiliation: Computational Chemistry and Biology Core Facility, Skaggs School of Pharmacy and Pharmaceutical Sciences, University of Colorado Anschutz Medical Campus, Aurora, CO 80045, USA.
authors.forename: Donald S
authors.initials: DS
authors.lastname: Backos
authors.affiliation: Division of Medical Genetics, Department of Pediatrics, Université de Montréal and CHU Sainte-Justine, 3175 Côte Sainte-Catherine, Montreal, QC H3T 1C5, Canada; College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi 712100, China.
authors.forename: Hao
authors.initials: H
authors.lastname: Yang
authors.affiliation: Division of Medical Genetics, Department of Pediatrics, Université de Montréal and CHU Sainte-Justine, 3175 Côte Sainte-Catherine, Montreal, QC H3T 1C5, Canada.
authors.forename: Grant A
authors.initials: GA
authors.lastname: Mitchell
authors.affiliation: Sarah W. Stedman Nutrition and Metabolism Center, Duke Molecular Physiology Institute, Duke University Medical Center, Durham, NC 27710, USA.
authors.forename: Olga R
authors.initials: OR
authors.lastname: Ilkayeva
authors.affiliation: Sarah W. Stedman Nutrition and Metabolism Center, Duke Molecular Physiology Institute, Duke University Medical Center, Durham, NC 27710, USA.
authors.forename: Robert D
authors.initials: RD
authors.lastname: Stevens
authors.affiliation: Sarah W. Stedman Nutrition and Metabolism Center, Duke Molecular Physiology Institute, Duke University Medical Center, Durham, NC 27710, USA.
authors.forename: Paul A
authors.initials: PA
authors.lastname: Grimsrud
authors.affiliation: Sarah W. Stedman Nutrition and Metabolism Center, Duke Molecular Physiology Institute, Duke University Medical Center, Durham, NC 27710, USA; Division of Endocrinology, Metabolism, and Nutrition, Department of Medicine, Duke University Medical Center, Durham, NC 27710, USA; Department of Pharmacology & Cancer Biology, Duke University Medical Center, Durham, NC 27710, USA. Electronic address: matthew.hirschey@duke.edu.
authors.forename: Matthew D
authors.initials: MD
authors.lastname: Hirschey
mesh_terms: D000214:Acyl Coenzyme A / Q000378:metabolism*; D000215:Acylation; D000812:Anhydrides / Q000378:metabolism; D055162:Biocatalysis; D002952:Citric Acid Cycle; D008239:Lysine / Q000378:metabolism; D055442:Metabolome; D008928:Mitochondria / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000378:metabolism*; D040901:Proteomics
chemical_list: D000214:Acyl Coenzyme A; D000812:Anhydrides; D011506:Proteins; C008047:3-hydroxy-3-methylglutaryl-coenzyme A; C015901:glutaryl-coenzyme A; C012046:succinyl-coenzyme A; D008239:Lysine
keywords: acyl-CoA; chemical biology; non-enzymatic; post-translational modifications; protein acylation; sirtuins