Protein kinase A downregulates the phosphorylation of p47 phox in human neutrophils: a possible pathway for inhibition of the respiratory burst.
Cyclic adenosine monophosphate (cAMP)-dependent protein kinase A(PKA) is considered to be a physiologic modulator of superoxide generation by stimulated neutrophils. Mechanisms of the inhibitory action of PKA are poorly understood. In this study, we investigated effects of cAMP-elevating agents on the phosphorylation of p47 phox in human neutrophils stimulated with the chemotactic peptide fMet-Leu-Phe (fMLP). We observed that the fMLP-induced phosphorylation of p47 phox, an essential component of neutrophil NADPH oxidase, was significantly attenuated in the presence of dibutyryl-cAMP or of receptor agonists of adenylate cyclase. This attenuation was reversed in the presence of 0.4 microM KT 5720, a selective inhibitor of PKA. The effects of cAMP agonists and of KT 5720 on the phosphorylation of p47 phox were paralleled by similar effects on superoxide generation. In neutrophils stimulated with phorbol myristate acetate (PMA), which directly activates protein kinase C (PKC), neither cAMP agonists nor dibutyryl cAMP exerted any effects on p47 phox phosphorylation or superoxide generation. These results indicated that the PKA-dependent downregulation of fMLP-induced p47 phox phosphorylation apparently involves step(s) in the fMLP-signaling pathway that are upstream of PKC. The inhibition demonstrated here of p47 phox phosphorylation by cAMP agonists may underlie a physiologically significant mechanism whereby cAMP modulates the receptor-mediated respiratory burst in neutrophils.
Bengis-Garber
C
C
Department of Biochemistry and Biochemical, Carmel Medical Center, Haifa, Israel.
Gruener
N
N
D003994:Bucladesine / Q000494:pharmacology; D002227:Carbazoles; D000242:Cyclic AMP / Q000819:agonists / Q000494:pharmacology; D017868:Cyclic AMP-Dependent Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D015536:Down-Regulation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006801:Humans; D007211:Indoles / Q000494:pharmacology; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D019255:NADPH Oxidases / Q000378:metabolism; D018375:Neutrophil Activation; D009504:Neutrophils / Q000378:metabolism*; D010750:Phosphoproteins / Q000378:metabolism*; D010766:Phosphorylation; D011758:Pyrroles / Q000494:pharmacology; D016897:Respiratory Burst; D015398:Signal Transduction; D013481:Superoxides / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D002227:Carbazoles; D004791:Enzyme Inhibitors; D007211:Indoles; D010750:Phosphoproteins; D011758:Pyrroles; D013481:Superoxides; C057416:KT 5720; D009240:N-Formylmethionine Leucyl-Phenylalanine; D003994:Bucladesine; D000242:Cyclic AMP; D019255:NADPH Oxidases; C060505:neutrophil cytosolic factor 1; D017868:Cyclic AMP-Dependent Protein Kinases; D013755:Tetradecanoylphorbol Acetate