2. Academic Validation
  3. Discovering Targets of Non-enzymatic Acylation by Thioester Reactivity Profiling

Discovering Targets of Non-enzymatic Acylation by Thioester Reactivity Profiling

  • Cell Chem Biol. 2017 Feb 16;24(2):231-242. doi: 10.1016/j.chembiol.2017.01.002.
Rhushikesh A Kulkarni 1,  Andrew J Worth 2,  Thomas T Zengeya 1,  Jonathan H Shrimp 1,  Julie M Garlick 1,  Allison M Roberts 1,  David C Montgomery 1,  Carole Sourbier 3,  Benjamin K Gibbs 3,  Clementina Mesaros 2,  Yien Che Tsai 4,  Sudipto Das 5,  King C Chan 5,  Ming Zhou 5,  Thorkell Andresson 5,  Allan M Weissman 4,  W Marston Linehan 3,  Ian A Blair 2,  Nathaniel W Snyder 6,  Jordan L Meier 7
Affiliations

Affiliations

  • 1 Chemical Biology Laboratory, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Frederick, MD 21702, USA.
  • 2 Penn SRP Center, Center for Excellence in Environmental Toxicology, University of Pennsylvania, Philadelphia, PA 19104, USA.
  • 3 Urologic Oncology Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20817, USA.
  • 4 Laboratory of Protein Dynamics and Signaling, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Frederick, MD 21702, USA.
  • 5 Protein Characterization Laboratory, Cancer Research Technology Program, Frederick National Laboratory for Cancer Research, Leidos Biomedical Research, Inc., Frederick, MD 21702, USA.
  • 6 Drexel University, A.J. Drexel Autism Institute, 3020 Market Street, Philadelphia, PA 19104, USA.
  • 7 Chemical Biology Laboratory, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Frederick, MD 21702, USA. Electronic address: [email protected].
PMID: 28163016 DOI: 10.1016/j.chembiol.2017.01.002
Abstract

Non-enzymatic protein modification driven by thioester reactivity is thought to play a major role in the establishment of cellular lysine acylation. However, the specific protein targets of this process are largely unknown. Here we report an experimental strategy to investigate non-enzymatic acylation in cells. Specifically, we develop a chemoproteomic method that separates thioester reactivity from enzymatic utilization, allowing selective enrichment of non-enzymatic acylation targets. Applying this method to Cancer cell lines identifies numerous candidate targets of non-enzymatic acylation, including several enzymes in lower glycolysis. Functional studies highlight malonyl-CoA as a reactive thioester metabolite that can modify and inhibit glycolytic Enzyme activity. Finally, we show that synthetic thioesters can be used as novel reagents to probe non-enzymatic acylation in living cells. Our studies provide new insights into the targets and drivers of non-enzymatic acylation, and demonstrate the utility of reactivity-based methods to experimentally investigate this phenomenon in biology and disease.

Keywords

Warburg effect; acetylation; acylation; epigenetics; glycolysis; malonylation; metabolism; non-enzymatic; reactivity-based protein profiling; thioester.

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