Protein kinase C-beta contributes to NADPH oxidase activation in neutrophils.
We have analysed the involvement of the beta isotype of the protein kinase C (PKC) family in the activation of NADPH oxidase in primary neutrophils. Using immunofluorescence and cell fractionation, PKC-beta is shown to be recruited to the plasma membrane upon stimulation with phorbol ester and to the phagosomal membrane upon phagocytosis of IgG-coated particles (Fcgamma-receptor stimulus). The time course of recruitment is similar to that of NADPH oxidase activation by these stimuli. The PKC-beta specific inhibitor 379196 inhibits the response to PMA as well as to IgG-coated bacteria. Partial inhibition occurs between 10 and 100 nM of inhibitor, the concentration at which PKC-beta, but not other PKC isotypes, is targeted. Neutrophils isolated from a mouse that lacks PKC-beta also showed an inhibition of NADPH oxidase activation by PMA and IgG-coated particles. The level of inhibition is comparable to that achieved with 379196 in human neutrophils. Thus the PKC-beta isotype mediates activation of NADPH oxidase by PMA and by stimulation of Fcgamma receptors in neutrophils.
Dekker
L V
LV
Centre for Molecular Medicine, University College London, 5 University Street, London WC1E 6JJ, U.K.
Leitges
M
M
Altschuler
G
G
Mistry
N
N
McDermott
A
A
Roes
J
J
Segal
A W
AW
D000818:Animals; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006801:Humans; D007074:Immunoglobulin G; D066298:In Vitro Techniques; D007211:Indoles / Q000494:pharmacology; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000097:blood*; D007700:Kinetics; D007840:Latex; D008301:Maleimides / Q000494:pharmacology; D051379:Mice; D019255:NADPH Oxidases / Q000097:blood*; D009504:Neutrophils / Q000201:enzymology / Q000502:physiology*; D010587:Phagocytosis; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000097:blood*; D064546:Protein Kinase C beta; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D004791:Enzyme Inhibitors; D007074:Immunoglobulin G; D007211:Indoles; D007527:Isoenzymes; D007840:Latex; D008301:Maleimides; C408699:compound 371196; D019255:NADPH Oxidases; D011493:Protein Kinase C; D064546:Protein Kinase C beta; D013755:Tetradecanoylphorbol Acetate