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PMID: 20519126 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Use of cells expressing gamma subunit variants to identify diverse mechanisms of AMPK activation.

Cell metabolism ·Vol. 11 ·No. 6 ·2010-06-09 ·Pages 554-65

Hawley SA, Ross FA, Chevtzoff C, Green KA, Evans A, Fogarty S, Towler MC, Brown LJ, Ogunbayo OA, Evans AM, Hardie DG

Abstract

A wide variety of agents activate AMPK, but in many cases the mechanisms remain unclear. We generated isogenic cell lines stably expressing AMPK complexes containing AMP-sensitive (wild-type, WT) or AMP-insensitive (R531G) gamma2 variants. Mitochondrial poisons such as oligomycin and dinitrophenol only activated AMPK in WT cells, as did AICAR, 2-deoxyglucose, hydrogen peroxide, metformin, phenformin, galegine, troglitazone, phenobarbital, resveratrol, and berberine. Excluding AICAR, all of these also inhibited cellular energy metabolism, shown by increases in ADP:ATP ratio and/or by decreases in cellular oxygen uptake measured using an extracellular flux analyzer. By contrast, A769662, the Ca(2+) ionophore, A23187, osmotic stress, and quercetin activated both variants to varying extents. A23187 and osmotic stress also increased cytoplasmic Ca(2+), and their effects were inhibited by STO609, a CaMKK inhibitor. Our approaches distinguish at least six different mechanisms for AMPK activation and confirm that the widely used antidiabetic drug metformin activates AMPK by inhibiting mitochondrial respiration.

MeSH Terms
AMP-Activated Protein Kinases/antagonists & inhibitors,metabolism Adenosine Diphosphate/metabolism Adenosine Triphosphate/metabolism Anti-Bacterial Agents/pharmacology Benzimidazoles/pharmacology Calcimycin/pharmacology Calcium/metabolism Cell Line Dinitrophenols/pharmacology Energy Metabolism Enzyme Activation Humans Hypoglycemic Agents/pharmacology Metformin/pharmacology Naphthalimides/pharmacology Oligomycins/pharmacology Phosphorylation Protein Subunits/metabolism
Chemicals
Anti-Bacterial Agents Benzimidazoles Dinitrophenols Hypoglycemic Agents Naphthalimides Oligomycins Protein Subunits STO 609 Calcimycin Adenosine Diphosphate Adenosine Triphosphate Metformin AMP-Activated Protein Kinases Calcium
Authors & Affiliations
11 authors, click to expand affiliations / ORCID
Hawley Simon A
Division of Molecular Physiology, College of Life Sciences, University of Dundee, Dundee DD1 5EH, Scotland, UK.
Ross Fiona A
Chevtzoff Cyrille
Green Kevin A
Evans Ashleigh
Fogarty Sarah
Towler Mhairi C
Brown Laura J
Ogunbayo Oluseye A
Evans A Mark
Hardie D Grahame
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Article Info
Journal
Cell metabolism
Abbr.
Cell Metab
ISSN
1932-7420
DOI
10.1016/j.cmet.2010.04.001
Published
2010-06-09
Pages
554-65
Language
English
Region
United States
NLM ID
101233170
PMCID
PMC2935965
Subset
IM
External Links
PubMed source DOI: 10.1016/j.cmet.2010.04.001 PMC Full Text View journal details
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