The stimulation of glycolysis by hypoxia in activated monocytes is mediated by AMP-activated protein kinase and inducible 6-phosphofructo-2-kinase.
The activation of monocytes involves a stimulation of glycolysis, release of potent inflammatory mediators, and alterations in gene expression. All of these processes are known to be further increased under hypoxic conditions. The activated monocytes express inducible 6-phosphofructo-2-kinase (iPFK-2), which synthesizes fructose 2,6-bisphosphate, a stimulator of glycolysis. During ischemia, AMP-activated protein kinase (AMPK) activates the homologous heart 6-phosphofructo-2-kinase isoform by phosphorylating its Ser-466. Here, we studied the involvement of AMPK and iPFK-2 in the stimulation of glycolysis in activated monocytes under hypoxia. iPFK-2 was phosphorylated on the homologous serine (Ser-461) and activated by AMPK in vitro. The activation of human monocytes by lipopolysaccharide induced iPFK-2 expression and increased fructose 2,6-bisphosphate content and glycolysis. The incubation of activated monocytes with oligomycin, an inhibitor of oxidative phosphorylation, or under hypoxic conditions activated AMPK and further increased iPFK-2 activity, fructose 2,6-bisphosphate content, and glycolysis. In cultured human embryonic kidney 293 cells, the expression of a dominant-negative AMPK prevented both the activation and phosphorylation of co-transfected iPFK-2 by oligomycin. It is concluded that the stimulation of glycolysis by hypoxia in activated monocytes requires the phosphorylation and activation of iPFK-2 by AMPK.
Marsin
Anne-Sophie
AS
Hormone and Metabolic Research Unit, University of Louvain Medical School and Christian de Duve International Institute of Cellular and Molecular Pathology, Avenue Hippocrate 75, B-1200 Brussels, Belgium.
Bouzin
Caroline
C
Bertrand
Luc
L
Hue
Louis
L
D055372:AMP-Activated Protein Kinases; D015687:Cell Hypoxia; D002478:Cells, Cultured; D004789:Enzyme Activation; D006019:Glycolysis; D006801:Humans; D008070:Lipopolysaccharides / Q000494:pharmacology; D009000:Monocytes / Q000378:metabolism*; D009097:Multienzyme Complexes / Q000502:physiology*; D009840:Oligomycins / Q000494:pharmacology; D025481:Phosphofructokinase-2 / Q000502:physiology*; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000502:physiology*
D008070:Lipopolysaccharides; D009097:Multienzyme Complexes; D009840:Oligomycins; D025481:Phosphofructokinase-2; D017346:Protein Serine-Threonine Kinases; D055372:AMP-Activated Protein Kinases