The PGE2-induced inhibition of the PLD activation pathway stimulated by fMLP in human neutrophils is mediated by PKA at the PI3-Kgamma level.
Prostaglandin E2 (PGE2), an eicosanoid that modulates inflammation, inhibits several chemoattractant-elicited functions in neutrophils such as chemotaxis, production of superoxide anions, adhesion, secretion of cytotoxic enzymes and synthesis of leukotriene B4. We previously reported that PGE2 inhibits the fMLP signaling pathway that leads to PLD activation through suppression of PI3-Kgamma activity and the decreased recruitment to membranes of PLD activation factors, PKC, Rho and Arf-GTPases. This effect is mediated via the EP2 receptors known to raise cAMP in cells. The inhibition of most fMLP-induced functional responses by PGE2 via EP2 receptors is mediated by PKA, except the chemotactic response. We have investigated the role of PKA in the EP2-mediated inhibition of the PLD activation pathway. H-89, a selective PKA pharmacological inhibitor suppressed the inhibitory effects of PGE2 at all stages of the PLD pathway activated by fMLP, i.e. PLD activity, translocation to membranes of PKCalpha, Rho and Arf-GTPases, calcium influx, tyrosine phosphorylation of proteins and finally translocation of p110gamma catalytic subunit of PI3-K to membranes. However, neither PLD nor PI3-Kgamma was substrate of PKA. These data provide evidence that PGE2-stimulated PKA activity regulates the PLD pathway stimulated by fMLP at the level of PI3-Kgamma and that the inhibition of PI3-Kgamma activation by PKA is a complex mechanism that remains to be completely elucidated.
Burelout
Chantal
C
Centre de Recherche en Rhumatologie-Immunologie, Centre de Recherche du Centre Hospitalier Universitaire de Québec, Département d'Anatomie-Physiologie, Québec, Canada.
Thibault
Nathalie
N
Harbour
Danielle
D
Naccache
Paul H
PH
Bourgoin
Sylvain G
SG
D002118:Calcium / Q000378:metabolism; D002478:Cells, Cultured; D058544:Class Ib Phosphatidylinositol 3-Kinase; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D015232:Dinoprostone / Q000494:pharmacology*; D004789:Enzyme Activation / Q000187:drug effects; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology*; D009504:Neutrophils / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism*; D010739:Phospholipase D / Q000037:antagonists & inhibitors* / Q000378:metabolism; D010766:Phosphorylation; D018078:Receptors, Prostaglandin E / Q000378:metabolism; D058307:Receptors, Prostaglandin E, EP2 Subtype
D007527:Isoenzymes; C545644:PTGER2 protein, human; D018078:Receptors, Prostaglandin E; D058307:Receptors, Prostaglandin E, EP2 Subtype; D009240:N-Formylmethionine Leucyl-Phenylalanine; D019869:Phosphatidylinositol 3-Kinases; D058544:Class Ib Phosphatidylinositol 3-Kinase; C546760:PIK3CG protein, human; D017868:Cyclic AMP-Dependent Protein Kinases; D010739:Phospholipase D; D015232:Dinoprostone; D002118:Calcium