Exposure to hydrogen peroxide induces oxidation and activation of AMP-activated protein kinase.
Although metabolic conditions associated with an increased AMP/ATP ratio are primary factors in the activation of 5'-adenosine monophosphate-activated protein kinase (AMPK), a number of recent studies have shown that increased intracellular levels of reactive oxygen species can stimulate AMPK activity, even without a decrease in cellular levels of ATP. We found that exposure of recombinant AMPKαβγ complex or HEK 293 cells to H(2)O(2) was associated with increased kinase activity and also resulted in oxidative modification of AMPK, including S-glutathionylation of the AMPKα and AMPKβ subunits. In experiments using C-terminal truncation mutants of AMPKα (amino acids 1-312), we found that mutation of cysteine 299 to alanine diminished the ability of H(2)O(2) to induce kinase activation, and mutation of cysteine 304 to alanine totally abrogated the enhancing effect of H(2)O(2) on kinase activity. Similar to the results obtained with H(2)O(2)-treated HEK 293 cells, activation and S-glutathionylation of the AMPKα subunit were present in the lungs of acatalasemic mice or mice treated with the catalase inhibitor aminotriazole, conditions in which intracellular steady state levels of H(2)O(2) are increased. These results demonstrate that physiologically relevant concentrations of H(2)O(2) can activate AMPK through oxidative modification of the AMPKα subunit. The present findings also imply that AMPK activation, in addition to being a response to alterations in intracellular metabolic pathways, is directly influenced by cellular redox status.
Zmijewski
Jaroslaw W
JW
From the Department of Medicine, Birmingham, Alabama 35294-0012; Center for Free Radical Biology, University of Alabama at Birmingham, Birmingham, Alabama 35294-0012. Electronic address: zmijewsk@uab.edu.
Banerjee
Sami
S
From the Department of Medicine, Birmingham, Alabama 35294-0012.
Bae
Hongbeom
H
From the Department of Medicine, Birmingham, Alabama 35294-0012; Department of Anesthesiology and Pain Medicine, Chonnam National University Medical School, Gwangju 501-746, Republic of Korea.
Friggeri
Arnaud
A
From the Department of Medicine, Birmingham, Alabama 35294-0012.
Lazarowski
Eduardo R
ER
Department of Medicine, University of North Carolina, Chapel Hill, North Carolina 27599-7248.
Abraham
Edward
E
From the Department of Medicine, Birmingham, Alabama 35294-0012; Center for Free Radical Biology, University of Alabama at Birmingham, Birmingham, Alabama 35294-0012. Electronic address: eabraham@uab.edu.
D055372:AMP-Activated Protein Kinases / Q000235:genetics / Q000378:metabolism*; D020642:Acatalasia / Q000235:genetics / Q000378:metabolism; D000249:Adenosine Monophosphate / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism*; D000640:Amitrole / Q000494:pharmacology; D000818:Animals; D002374:Catalase / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D020134:Catalytic Domain; D002460:Cell Line; D004790:Enzyme Induction / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005978:Glutathione / Q000378:metabolism; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D051379:Mice; D009154:Mutation; D016877:Oxidants / Q000494:pharmacology*; D010084:Oxidation-Reduction / Q000187:drug effects; D011499:Protein Processing, Post-Translational / Q000187:drug effects*
D004791:Enzyme Inhibitors; D016877:Oxidants; D000249:Adenosine Monophosphate; D000255:Adenosine Triphosphate; D006861:Hydrogen Peroxide; D002374:Catalase; D055372:AMP-Activated Protein Kinases; D005978:Glutathione; D000640:Amitrole