Chemical genetic screen identifies Gapex-5/GAPVD1 and STBD1 as novel AMPK substrates

TY - JOUR

T1 - Chemical genetic screen identifies Gapex-5/GAPVD1 and STBD1 as novel AMPK substrates

AU - Ducommun, Serge

AU - Deak, Maria

AU - Zeigerer, Anja

AU - Göransson, Olga

AU - Seitz, Susanne

AU - Collodet, Caterina

AU - Madsen, Agnete Bjerregaard

AU - Jensen, Thomas Elbenhardt

AU - Viollet, Benoit

AU - Foretz, Marc

AU - Gut, Philipp

AU - Sumpton, David

AU - Sakamoto, Kei

N1 - CURIS 2019 NEXS 073 Copyright © 2019. Published by Elsevier Inc.

PY - 2019/5

Y1 - 2019/5

N2 - 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.

AB - 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.

KW - Faculty of Science

KW - GTPase activating protein and VPS9 domains 1

KW - Shokat

KW - Starch-binding domain 1

U2 - 10.1016/j.cellsig.2019.02.001

DO - 10.1016/j.cellsig.2019.02.001

M3 - Journal article

C2 - 30772465

SN - 0898-6568

VL - 57

SP - 45

EP - 57

JO - Cellular Signalling

JF - Cellular Signalling

ER -