Mechanism of action of compound-13: an α1-selective small molecule activator of AMPK

  • Chem Biol. 2014 Jul 17;21(7):866-79. doi: 10.1016/j.chembiol.2014.05.014.
Roger W Hunter  1 ,  Marc Foretz  2 ,  Laurent Bultot  3 ,  Morgan D Fullerton  4 ,  Maria Deak  3 ,  Fiona A Ross  5 ,  Simon A Hawley  5 ,  Natalia Shpiro  6 ,  Benoit Viollet  2 ,  Denis Barron  3 ,  Bruce E Kemp  7 ,  Gregory R Steinberg  4 ,  D Grahame Hardie  5 ,  Kei Sakamoto  8
Affiliations
  • 1. MRC Protein Phosphorylation and Ubiquitylation Unit, College of Life Sciences, University of Dundee, Dow Street, Dundee, DD1 5EH Scotland, UK; Nestlé Institute of Health Sciences SA, EPFL Innovation Park, bâtiment G, 1015 Lausanne, Switzerland.
  • 2. Inserm, U1016, Institut Cochin, 24 rue du Faubourg Saint-Jacques, 75014 Paris, France; CNRS, UMR8104, Paris, France; Université Paris Descartes, Sorbonne Paris cité, 75006 Paris, France.
  • 3. Nestlé Institute of Health Sciences SA, EPFL Innovation Park, bâtiment G, 1015 Lausanne, Switzerland.
  • 4. Division of Endocrinology and Metabolism, Department of Medicine, McMaster University, 1280 Main West Street, Hamilton ON L8N 3Z5, Canada.
  • 5. Division of Cell Signalling and Immunology, College of Life Sciences, University of Dundee, Dundee DD1 5EH, UK.
  • 6. MRC Protein Phosphorylation and Ubiquitylation Unit, College of Life Sciences, University of Dundee, Dow Street, Dundee, DD1 5EH Scotland, UK.
  • 7. Protein Chemistry and Metabolism, St. Vincent's Institute and Department of Medicine, University of Melbourne, 41 Victoria Parade, Fitzroy VIC 3065, Australia.
  • 8. MRC Protein Phosphorylation and Ubiquitylation Unit, College of Life Sciences, University of Dundee, Dow Street, Dundee, DD1 5EH Scotland, UK; Nestlé Institute of Health Sciences SA, EPFL Innovation Park, bâtiment G, 1015 Lausanne, Switzerland. Electronic address: [email protected].
PMID: 25036776 DOI: 10.1016/j.chembiol.2014.05.014
Abstract

AMPK is a sensor of cellular energy status and a promising target for drugs aimed at metabolic disorders. We have studied the selectivity and mechanism of a recently described activator, C2, and its cell-permeable prodrug, C13. C2 was a potent allosteric activator of α1-complexes that, like AMP, also protected against Thr172 dephosphorylation. Compared with AMP, C2 caused only partial allosteric activation of α2-complexes and failed to protect them against dephosphorylation. We show that both effects could be fully restored by exchanging part of the linker between the autoinhibitory and C-terminal domains in α2, containing the equivalent region from α1 thought to interact with AMP bound in site 3 of the γ subunit. Consistent with our results in cell-free assays, C13 potently inhibited lipid synthesis in hepatocytes from wild-type and was largely ineffective in AMPK-knockout hepatocytes; its effects were more severely affected by knockout of α1 than of α2, β1, or β2.