1. Chemical genetic screen identifies Gapex-5/GAPVD1 and STBD1 as novel AMPK substrates
- Author
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Olga Göransson, Susanne Seitz, Philipp Gut, Maria Deak, Anja Zeigerer, Agnete B. Madsen, Marc Foretz, Benoit Viollet, David Sumpton, Thomas E. Jensen, Kei Sakamoto, Caterina Collodet, Serge Ducommun, Nestlé Institute of Health Sciences SA [Lausanne, Switzerland], Max Planck Institute of Molecular Cell Biology and Genetics (MPI-CBG), Max-Planck-Gesellschaft, Department of Experimental Medical Sciences [Lund], Lund University [Lund], German Research Center for Environmental Health - Helmholtz Center München (GmbH), Department of Nutrition, Exercise and Sports [Copenhagen], Faculty of Science [Copenhagen], University of Copenhagen = Københavns Universitet (KU)-University of Copenhagen = Københavns Universitet (KU), Institut Cochin (IC UM3 (UMR 8104 / U1016)), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), and Cancer Research UK Beatson Institute [Glasgow]
- Subjects
0301 basic medicine ,Regulator ,Muscle Proteins ,AMP-Activated Protein Kinases ,Mass Spectrometry ,Substrate Specificity ,03 medical and health sciences ,0302 clinical medicine ,Faculty of Science ,Animals ,Guanine Nucleotide Exchange Factors ,Homeostasis ,Phosphorylation ,[SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM] ,Protein kinase A ,Mice, Knockout ,Effector ,Activator (genetics) ,Chemistry ,Starch-binding domain 1 ,Membrane Proteins ,AMPK ,[SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology ,Shokat ,Cell Biology ,[SDV.MHEP.EM]Life Sciences [q-bio]/Human health and pathology/Endocrinology and metabolism ,Lipid Metabolism ,[SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM] ,Cell biology ,030104 developmental biology ,Liver ,GTPase activating protein and VPS9 domains 1 ,030220 oncology & carcinogenesis ,Hepatocytes ,Genetic screen - Abstract
International audience; AMP-activated protein kinase (AMPK) is a key regulator of cellular energy homeostasis, acting as a sensor of energy and nutrient status. As such, AMPK is considered a promising drug target for treatment of medical conditions particularly associated with metabolic dysfunctions. To better understand the downstream effectors and physiological consequences of AMPK activation, we have employed a chemical genetic screen in mouse primary hepatocytes in an attempt to identify novel AMPK targets. Treatment of hepatocytes with a potent and specific AMPK activator 991 resulted in identification of 65 proteins phosphorylated upon AMPK activation, which are involved in a variety of cellular processes such as lipid/glycogen metabolism, vesicle trafficking, and cytoskeleton organisation. Further characterisation and validation using mass spectrometry followed by immunoblotting analysis with phosphorylation site-specific antibodies identified AMPK-dependent phosphorylation of Gapex-5 (also known as GTPase-activating protein and VPS9 domain-containing protein 1 (GAPVD1)) on Ser902 in hepatocytes and starch-binding domain 1 (STBD1) on Ser175 in multiple cells/tissues. As new promising roles of AMPK as a key metabolic regulator continue to emerge, the substrates we identified could provide new mechanistic and therapeutic insights into AMPK-activating drugs in the liver.
- Published
- 2019