Activation of Protein Kinase C by Respiratory Burst Stimulants Desensitises β2-Adrenoceptors on Human Neutrophils

1991 ◽  
Vol 3 (S2) ◽  
pp. 54-65 ◽  
Author(s):  
Maurizio Bevilacqua ◽  
G. Norbiato ◽  
G. Baldi ◽  
P. Bertora ◽  
T. Vago ◽  
...  
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Respiratory Burst ◽  
Human Neutrophils ◽  
Kinase C

scholarly journals Protein kinase C activity is not involved in N-formylmethionyl-leucyl-phenylalanine-induced phospholipase D activation in human neutrophils, but is essential for concomitant NADPH oxidase activation: studies with a staurosporine analogue with improved selectivity for protein kinase C

1993 ◽  
Vol 292 (3) ◽  
pp. 781-785 ◽  
Author(s):  
G C Kessels ◽  
K H Krause ◽  
A J Verhoeven
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Cyclic Amp ◽  
Nadph Oxidase ◽  
Phospholipase D ◽  
Respiratory Burst ◽  
Feedback Inhibition ◽  
Human Neutrophils ◽  
Kinase C ◽  
Pkc Activation

Stimulation of human neutrophils by the receptor agonist N-formylmethionyl-leucyl-phenylalanine (fMLP) results in a respiratory burst, catalysed by an NADPH oxidase. Concomitantly, phospholipase D (PLD) is activated. To investigate the role of protein kinase C (PKC) in these neutrophil responses, we have compared the effects of staurosporine and a structural analogue of staurosporine (cgp41251), that reflects a higher selectivity towards PKC [Meyer, Regenass, Fabbro, Alteri, Rösel, Müller, Caravatti and Matter (1989) Int. J. Cancer 43, 851-856]. Both staurosporine and cgp41251 dose-dependently inhibited the production of superoxide induced by phorbol 12-myristate 13-acetate (PMA). Both compounds also caused inhibition of the fMLP-induced respiratory burst, but with a lower efficacy during the initiation phase of this response. This latter observation cannot be taken as evidence against PKC involvement in the activation of the respiratory burst, because pretreatment of neutrophils with ionomycin before PMA stimulation also results in a lower efficacy of inhibition. Activation of PLD by fMLP was enhanced in the presence of staurosporine, but not in the presence of cgp41251. Enhancement of PLD activation was also observed in the presence of H-89, an inhibitor of cyclic-AMP-dependent protein kinase (PKA). Both staurosporine and H-89 reversed the dibutyryl-cyclic-AMP-induced inhibition of PLD activation, whereas cgp41251 was without effect. These results indicate that the potentiating effect of staurosporine on PLD activation induced by fMLP does not reflect a feedback inhibition by PKC activation, but instead a feedback inhibition by PKC activation. Taken together, our results indicate that in human neutrophils: (i) PKC activity is not essential for fMLP-induced activation of PLD; (ii) PKC activity does play an essential role in the activation of the respiratory burst by fMLP, other than mediating or modulating PLD activation; (iii) there exists a negative-feedback mechanism on fMLP-induced PLD activation by concomitant activation of PKA.

scholarly journals Early signalling events implicated in leukotriene B4-induced activation of the NADPH oxidase in eosinophils: role of Ca2+, protein kinase C and phospholipases C and D

1995 ◽  
Vol 310 (3) ◽  
pp. 795-806 ◽  
Author(s):  
R S Perkins ◽  
M A Lindsay ◽  
P J Barnes ◽  
M A Giembycz
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Nadph Oxidase ◽  
Respiratory Burst ◽  
Leukotriene B4 ◽  
Human Neutrophils ◽  
H2o2 Production ◽  
Intact Cells ◽  
Kinase C ◽  
H2o2 Generation

The early signalling events that may ultimately contribute to the assembly and subsequent activation of the NADPH oxidase in guinea-pig peritoneal eosinophils were investigated in response to leukotriene B4 (LTB4). LTB4 promoted a rapid, transient and receptor-mediated increase in the rate of H2O2 generation that was potentiated by R 59 022, a diradylglycerol (DRG) kinase inhibitor, implicating protein kinase C (PKC) in the genesis of this response. This conclusion was supported by the finding that the PKC inhibitor, Ro 31-8220, attenuated (by about 30%) the peak rate of LTB4-induced H2O2 generation under conditions where the same response evoked by 4 beta-phorbol 12,13-dibutyrate (PDBu) was inhibited by more than 90%. Paradoxically, Ro 31-8220 doubled the amount of H2O2 produced by LTB4 which may relate to the ability of PKC to inhibit cell signalling through phospholipase C (PLC). Indeed, Ro 31-8220 significantly enhanced LTB4-induced Ins(1,4,5)P3 accumulation and the duration of the Ca2+ transient in eosinophils. Experiments designed to assess the relative importance of DRG-mobilizing phospholipases in LTB4-induced oxidase activation indicated that phospholipase D (PLD) did not play a major role. Thus, although H2O2 generation was abolished by butan-1-ol, this was apparently unrelated to the inhibition of PLD, as LTB4 failed to stimulate the formation of Ptd[3H]BuOH in [3H]butan-1-ol-treated eosinophils. Rather, the inhibition was probably due to the ability of butan-1-ol to increase the eosinophil cyclic AMP content. In contrast, Ca(2+)- and PLC-driven mechanisms were implicated in H2O2 generation, as LTB4 elevated the Ins(1,4,5)P3 content and intracellular free Ca2+ concentration in intact cells, and cochelation of extracellular and intracellular Ca2+ significantly attenuated LTB4-induced H2O2 generation. Pretreatment of eosinophils with wortmannin did not affect LTB4-induced H2O2 production at concentrations at which it abolished the respiratory burst evoked by formylmethionyl-leucylphenylalanine in human neutrophils. Collectively, these data suggest that LTB4 activates the NADPH oxidase in eosinophils by PLD- and PtdIns 3-kinase-independent mechanisms that involve Ca2+, PLC and PKC. Furthermore, the activation of additional pathways that do not require Ca2+ is also suggested by the finding that LTB4 evoked a significant respiratory burst in Ca(2+)-depleted cells.

scholarly journals Chemotactic peptide-induced activation of respiratory burst in human neutrophils. Involvement of a protein kinase C-independent signaling pathway.

Ensho ◽  
1993 ◽  
Vol 13 (4) ◽  
pp. 325-329
Author(s):  
Eriko Azuma ◽  
Seiichi Kitagawa ◽  
Akira Yuo ◽  
Hideaki Mizoguchi ◽  
Fumimaro Takaku ◽  
...  
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Signaling Pathway ◽  
Respiratory Burst ◽  
Human Neutrophils ◽  
Chemotactic Peptide ◽  
Kinase C

scholarly journals Guanine nucleotides induce tyrosine phosphorylation and activation of the respiratory burst in neutrophils

1989 ◽  
Vol 257 (3) ◽  
pp. 893-897 ◽  
Author(s):  
P E Nasmith ◽  
G B Mills ◽  
S Grinstein
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Tyrosine Phosphorylation ◽  
Respiratory Burst ◽  
Tyrosine Phosphatase ◽  
Guanine Nucleotides ◽  
Human Neutrophils ◽  
Guanine Nucleotide ◽  
Phosphorylated Proteins ◽  
Kinase C

Activation of the NADPH oxidase was examined in electrically permeabilized human neutrophils exposed to non-hydrolysable guanine nucleotides. Guanosine 5'-[gamma-thio]triphosphate (GTP[S]) induced a marked increase in the rate of O2 consumption, which was partially resistant to staurosporine, an inhibitor of protein kinase C, under conditions where the response to diacylglycerol was virtually abolished. The respiratory burst elicited by GTP[S] was dependent on the presence of ATP and Mg2+, suggesting involvement of phosphorylation reactions. Accordingly, phosphoprotein formation was greatly stimulated by the guanine nucleotide. The polypeptide phosphorylation pattern induced by GTP[S] was similar to, but not identical with, that observed with diacylglycerol, indicating the activation of kinases other than protein kinase C by the guanine nucleotide. The possible involvement of tyrosine kinases was assessed by immunoblotting using anti-phosphotyrosine antibodies. Treatment of electroporated cells with GTP[S] stimulated the accumulation of tyrosine-phosphorylated proteins. This effect was not induced by diacylglycerol, indicating that tyrosine phosphorylation is not secondary to stimulation of protein kinase C. The results indicate that, in neutrophils, activated G-proteins can stimulate tyrosine kinase and/or inhibit tyrosine phosphatase activity. Changes in the amounts of tyrosine-phosphorylated proteins may signal activation of the respiratory burst.

scholarly journals Oxidative and adhesive responses of human neutrophils to nitrovasodilatorsin vitro: the role of protein kinases

2003 ◽  
Vol 12 (6) ◽  
pp. 345-353 ◽  
Author(s):  
Magdalena Klink ◽  
Henryk Tchorzewski ◽  
Zofia Sulowska
Keyword(s):  
Protein Kinase C ◽  
Heart Disease ◽  
Protein Kinase ◽  
Protein Kinases ◽  
Respiratory Burst ◽  
Protein Tyrosine Kinase ◽  
Inhibitory Effect ◽  
Human Neutrophils ◽  
No Donors ◽  
Kinase C

Background:Sodium nitroprusside (SNP) and molsidomine are used in the treatment of coronary heart disease. Since the neutrophils play a pathological role in ischaemic heart disease, it is important to understand the direct action of nitrovasodilators on their function.Aim:We examined the effects of SNP and 3-morpholinosydnonimine (SIN-1, molsidomine metabolite) on the respiratory burst of human neutrophils and their adhesionsin vitro. The influence of nitric oxide (NO) donors on the activity of protein kinases, which are involved in the NADPH oxidase activation, was also investigated.Methods:The respiratory burst of neutrophils was determined by chemiluminescence and fluorescence methods, while the adhesion was assayed by adherence of neutrophils to the plastic surface.Results:NO donors decreased the oxidative burst of activated neutrophils. However, the effects of SNP and SIN-1 strongly depended on the treatment time of neutrophils and on the stimulus employed to cells activation. Protein kinase C inhibitor did not prevent the inhibitory effect of SIN-1, but diminished the inhibitory effect of SNP on the neutrophils' respiratory burst. Protein tyrosine kinase inhibitor did not affect the action of SNP, but diminished the inhibitory effect of SIN-1 on fMLP-stimulated but not on PMA-stimulated oxidative burst of neutrophils. This suggests that SNP action is mainly associated with protein kinase C, while SIN-1 is associated with protein tyrosine kinase activity. We also found that SIN-1 but not SNP diminished the adhesive activity of neutrophils.Conclusions:Our data show that SIN-1 biological effect on some neutrophils activity is different from both spermine NONOate and SNP, and mainly depends on ONOO−, while SNP action is mediated by NO.

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