title: Mechanism of action of compound-13: an α1-selective small molecule activator of AMPK.
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.
authors.affiliation: 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.
authors.forename: Roger W
authors.initials: RW
authors.lastname: Hunter
authors.affiliation: 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.
authors.forename: Marc
authors.initials: M
authors.lastname: Foretz
authors.affiliation: Nestlé Institute of Health Sciences SA, EPFL Innovation Park, bâtiment G, 1015 Lausanne, Switzerland.
authors.forename: Laurent
authors.initials: L
authors.lastname: Bultot
authors.affiliation: Division of Endocrinology and Metabolism, Department of Medicine, McMaster University, 1280 Main West Street, Hamilton ON L8N 3Z5, Canada.
authors.forename: Morgan D
authors.initials: MD
authors.lastname: Fullerton
authors.affiliation: Nestlé Institute of Health Sciences SA, EPFL Innovation Park, bâtiment G, 1015 Lausanne, Switzerland.
authors.forename: Maria
authors.initials: M
authors.lastname: Deak
authors.affiliation: Division of Cell Signalling and Immunology, College of Life Sciences, University of Dundee, Dundee DD1 5EH, UK.
authors.forename: Fiona A
authors.initials: FA
authors.lastname: Ross
authors.affiliation: Division of Cell Signalling and Immunology, College of Life Sciences, University of Dundee, Dundee DD1 5EH, UK.
authors.forename: Simon A
authors.initials: SA
authors.lastname: Hawley
authors.affiliation: MRC Protein Phosphorylation and Ubiquitylation Unit, College of Life Sciences, University of Dundee, Dow Street, Dundee, DD1 5EH Scotland, UK.
authors.forename: Natalia
authors.initials: N
authors.lastname: Shpiro
authors.affiliation: 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.
authors.forename: Benoit
authors.initials: B
authors.lastname: Viollet
authors.affiliation: Nestlé Institute of Health Sciences SA, EPFL Innovation Park, bâtiment G, 1015 Lausanne, Switzerland.
authors.forename: Denis
authors.initials: D
authors.lastname: Barron
authors.affiliation: Protein Chemistry and Metabolism, St. Vincent's Institute and Department of Medicine, University of Melbourne, 41 Victoria Parade, Fitzroy VIC 3065, Australia.
authors.forename: Bruce E
authors.initials: BE
authors.lastname: Kemp
authors.affiliation: Division of Endocrinology and Metabolism, Department of Medicine, McMaster University, 1280 Main West Street, Hamilton ON L8N 3Z5, Canada.
authors.forename: Gregory R
authors.initials: GR
authors.lastname: Steinberg
authors.affiliation: Division of Cell Signalling and Immunology, College of Life Sciences, University of Dundee, Dundee DD1 5EH, UK.
authors.forename: D Grahame
authors.initials: DG
authors.lastname: Hardie
authors.affiliation: 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: kei.sakamoto@rd.nestle.com.
authors.forename: Kei
authors.initials: K
authors.lastname: Sakamoto
mesh_terms: D055372:AMP-Activated Protein Kinases / Q000737:chemistry / Q000378:metabolism*; D000249:Adenosine Monophosphate / Q000494:pharmacology; D000595:Amino Acid Sequence; D000818:Animals; D004789:Enzyme Activation / Q000187:drug effects; D020536:Enzyme Activators / Q000378:metabolism / Q000494:pharmacology*; D004951:Esterification / Q000187:drug effects; D005227:Fatty Acids / Q000378:metabolism; D022781:Hepatocytes / Q000166:cytology / Q000187:drug effects; D050155:Lipogenesis / Q000187:drug effects; D051379:Mice; D008969:Molecular Sequence Data; D011355:Prodrugs / Q000378:metabolism / Q000494:pharmacology; D021122:Protein Subunits / Q000819:agonists / Q000737:chemistry / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D054852:Small Molecule Libraries / Q000378:metabolism / Q000494:pharmacology*; D013379:Substrate Specificity
chemical_list: D020536:Enzyme Activators; D005227:Fatty Acids; D011355:Prodrugs; D021122:Protein Subunits; D011994:Recombinant Proteins; D054852:Small Molecule Libraries; D000249:Adenosine Monophosphate; D055372:AMP-Activated Protein Kinases