Involvement of protein phosphatase 2A in PKC-independent pathway of neutrophil superoxide generation by fMLP.
We examined the effects of okadaic acid, a protein phosphatase 1 and 2A inhibitor, on superoxide generation in human neutrophils. Superoxide generation induced by fMLP was inhibited by low-dose okadaic acid (10-100 nM), but it had no effect on superoxide synthesis by PMA, and the fMLP-induced rise of the intracellular Ca2+ concentration was not affected by low-dose okadaic acid. These findings suggested that the inhibitory mechanism of okadaic acid might involve PKC-independent and Ca(2+)-independent pathways in fMLP induced NADPH oxidase activation. Both fMLP-stimulated phosphorylation of serine residues in p47phox and its translocation to the plasma membrane were suppressed by low-dose okadaic acid. On the other hand, PMA-induced phosphorylation and translocation of p47phox were not affected by such a low dose of okadaic acid. These findings suggested that fMLP induced phosphorylation of serine residues in p47phox was regulated by protein phosphatase 2A, and its phosphorylation was necessary for translocation and superoxide generation in fMLP-activated human neutrophils.
Okuyama
M
M
Department of Surgery II, Osaka University Medical School, Japan.
Sakon
M
M
Kambayashi
J
J
Kawasaki
T
T
Monden
M
M
D002118:Calcium / Q000097:blood; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004988:Ethers, Cyclic / Q000494:pharmacology; D006801:Humans; D066298:In Vitro Techniques; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology*; D009504:Neutrophils / Q000187:drug effects* / Q000378:metabolism; D019319:Okadaic Acid; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors / Q000502:physiology*; D010750:Phosphoproteins / Q000097:blood; D010766:Phosphorylation; D011493:Protein Kinase C / Q000502:physiology*; D054645:Protein Phosphatase 1; D054648:Protein Phosphatase 2; D013481:Superoxides / Q000097:blood*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D004791:Enzyme Inhibitors; D004988:Ethers, Cyclic; D010750:Phosphoproteins; C079789:neutrophil cytosol factor 67K; D013481:Superoxides; D019319:Okadaic Acid; D009240:N-Formylmethionine Leucyl-Phenylalanine; D011493:Protein Kinase C; D010749:Phosphoprotein Phosphatases; D054645:Protein Phosphatase 1; D054648:Protein Phosphatase 2; D013755:Tetradecanoylphorbol Acetate; D002118:Calcium