Abstract
Intact human neutrophils produced superoxide (O2 −) by the stimulation with N-formyl-methionyl-leucyl-phenylalanine (fMLP) even when the extracellular Ca2+ was absent (0.56±0.13 nmol/min per 106 cells). The production by fMLP was enhanced more than twice in the presence of the extracellular Ca2+. Moreover, the O2 − production by fMLP in the presence of extracellular Ca2+ was enhanced nearly three times by the treatment of cells with H-89, an inhibitor of cyclic AMP-dependent protein kinase (PKA). The enhancement was not observed when the extracellular Ca2+ was depleted from the reaction mixture. In addition, H-89 did not enhance fMLP-induced O2 − production of electropermeabilized neutrophils in which the intracellular Ca2+ concentration was fixed to about 100 nM. These observations suggest that not only Ca2+ influx but the inhibition of PKA is necessary for the maximum O2 − production by fMLP and that the O2 − production is partially suppressed by the activation of PKA induced by fMLP.
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References
Badwey JA, Kamovsky ML: Active oxygen species and the functions of phagocytic leukocytes. Ann Rev Biochem 49: 695–726, 1980
Rossi F: The O2 − forming NADPH oxidase of the phagocytes. Biochim Biophys Acta 853: 65–89, 1986
Morel F, Doussiere J, Vignais PD: The superoxide-generating oxidase of phagocytic cells. Eur J Biochem 201: 523–546, 1991
Agwu DE, McPhail LC, Chabot MC, Daniel LW, Wykle RL, McCall CE: Choline-linked phosphoglycerides. J Biol Chem 264: 1405–1413, 1989
Pai J-K, Siegel MI, Egan RW, Billah MM: Activation of phospholipase D by chemotactic peptide in HL-60 granulocytes. Biochem Biophys Res Commun 150: 355–364, 1988
Gelas P, Ribbes G, Record M, Terce F, Chap H: Differential activation by fMet-Leu-Phe and phorbol ester of a plasma membrane phosphatidylcholine-specific phospholipase D in human neutrophils. FEBS Lett 251: 213–218, 1989
Mullmann TJ, Siegel MI, Egan RW, Billah MM: Phorbol-12-myristate-13-acetate activation of phospholipase D in human neutrophils leads to the production of phosphatides and diglycerides. Biochem Biophys Res Commun 170: 1197–1202, 1990
Billah MM, Eckel S, Mullmann TJ, Egan RW, Siegel MI: Phosphatidylcholine hydrolysis by phospholipase D determines phosphatide and diglyceride levels in chemotactic peptide-stimulated human neutrophils. J Biol Chem 264: 17069–17077, 1989
Anthes JC, Eckel S, Siegel MI, Egan RW, Billah MM: Phospholipase D in homogenates from HL-60 granulocytes. Biochem Biophys Res Commun 163: 657–664, 1989
Agwu DE, McPhail LC, Sozzani S, Bass DA, McCall CE: Phosphatidic acid as a second messenger in human polymorphonuclear leukocytes. J Clin Invest 88: 531–539, 1991
Bonser RW, Thompson NT, Randall RW, Garland LG: Phospholipase D activation is functionally linked to superoxide generation in human neutrophils. Biochem J 264: 617–620, 1989
Rossi F, Grzeskowiak M, Della Bianca V, Calzetti F, Gandini G: Phosphatidic acid and not diacylglycerol generated by phospholipase D is functionally linked to the activation of the NADPH oxidase by FMLP in human neutrophils. Biochem Biophys Res Commun 168: 320–327, 1990
Bellavite P, Corso F, Dusi S, Grzeskowiak M, Della Bianca V, Rossi F: Activation of NADPH-dependent superoxide production in plasma membrane extracts of pig neutrophils by phosphatidic acid. J Biol Chem 263: 8210–8214, 1988
Mitsuyama T, Takeshige K, Minakami S: Phosphatidic acid induces the respiratory burst of electropermeabilized human neutrophils by acting on downstream step of protein kinase C. FEBS Lett 328: 67–70, 1993
Roger Kessels GC, Roos D, Verhoeven AJ: fMet-Leu-Phe-induced activation of phospholipase D in human neutrophils. J Biol Chem 266: 23152–23156, 1991
Mitsuyama T, Takeshige K, Minakami S: Cyclic AMP inhibits the respiratory burst of electropermeabilized human neutrophils at a downstream site of protein kinase C. Biochim Biophys Acta 1177: 167–173, 1993
Mitsuyama T, Takeshige K, Minakami S: Tyrosine phosphorylation is involved in the respiratory burst of electropermeabilized human neutrophils at a step before diacylglycerol formation by phospholipase C. FEBS Lett 322: 280–284, 1993
Wong K, Freund K: Inhibition of then-formylmethionyl-leucyl-phenylalanine induced respiratory burst in human neutrophils by adrenergic agonists and prostaglandins of the E series. Can J Physiol Pharmacol 59: 915–920, 1989
Fantone JC, Kinnes DA: Prostaglandin E1 and I2 modulation of superoxide production by human neutrophils. Biochem Biophys Res Commun 113: 506–512, 1983
Fujita I, Irita K, Takeshige K, Minakami S: Diacylglycerol, l-oleoyl-2-acetyl-glycerol, stimulates superoxide-generation from human neutrophils. Biochem Biophys Res Commun 120: 318–324, 1984
De Tongi P, Cabrini G, Di Virgilio F: Cyclic AMP inhibition of fMet-Leu-Phe-dependent metabolic responses in human neutrophils is not due to its effects on cytosolic Ca2+. Biochem J 224: 629–635, 1984
Tyagi SR, Olson SC, Burnham DN, Lambeth JD: Cyclic AMP-elevating agents block chemoatractant activation of diradylglycerol generation by inhibiting phospholipase D activation. J Biol Chem 266: 3498–3504, 1991
Chijiwa T, Mishima A, Hagiwara M, Sano M, Hayashi K, Inoue T, Naito K, Toshioka T, Hidaka H: Inhibition of forskolin-induced neurite outgrowth and protein phosphorylation by a newly synthesized selective outgrowth and protein phosphorylation by a newly synthesized selective inhibitor of cyclic AMP-dependent protein kinase. N-[2-(p-bromocinnamylamino)ethyl]-5-isoquinolinesulfonamide (H-89), of PC12D pheochromocytoma cells. J Biol Chem 265: 5267–5272, 1990
Nakagawara M, Takeshige K, Sumimoto H, Yoshitake J, Minakami S: Superoxide release and intracellular calcium of calcium-depleted human neutrophils stimulated by N-formyl-methionyl-leucyl-phenylalanine. Biochim Biophys Acta 805: 97–103, 1984
Kuroki M, Minakami S: Extracellular ATP triggers superoxide production in human neutrophils. Biochem Biophys Res Commun 162: 377–380, 1989
Lu DJ, Grinstein S: ATP and guanine nucleotide dependence of neutrophil activation. J Biol Chem 265: 13721–13729, 1990
Kakinuma K, Minakami S: Effects of fatty acids on superoxide radical generation in leukocytes. Biochim Biophys Acta 538: 50–59, 1978
Simchowitz L, Fischbein LC, Spillberg I, Atkinson JP: Induction of a transient elevation in intracellular levels of adenosine-3′, 5′-cyclic monophosphate by chemotactic factors. J Immunol 124: 1482–1491, 1980
Smolen JE, Weissmann G: Stimuli which provoke secretion of azurophil enzymes from human neutrophils induce increments in adenosine cyclic 3′–5′-monophosphate. Biochim Biophys Acta 672: 197–206, 1981
Hartfield PJ, Robinson JM: MgATP primes superoxide-generating responses in electropermeabilized neutrophils. Biochem Biophys Res Commun 170: 194–200, 1990
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Mitsuyama, T., Takeshige, K., Furuno, T. et al. An inhibitor of cyclic AMP-dependent protein kinase enhances the superoxide production of human neutrophils stimulated by N-formyl-methionyl-leucyl-phenylalanine. Mol Cell Biochem 145, 19–24 (1995). https://doi.org/10.1007/BF00925708
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DOI: https://doi.org/10.1007/BF00925708