Abstract
Phospholipase A2 (PLA2) inhibitors suppressed simultaneously, in a dose-dependent manner, the activation of NADPH oxidase and the release of 3H-labelled arachidonic acid ([3H]AA) stimulated by either phorbol 12-myristate 13-acetate (PMA) or opsonized zymosan (OZ) in human neutrophils. In spite of total inhibition of superoxide production in the presence of the PLA2 inhibitors, 10 microM bromophenacyl bromide (BPB) or 20 microM quinacrine, a maximal phosphorylation of p47 and translocation of p47 and p67 to the neutrophil membranes induced by PMA or OZ was observed. Addition of 10 microM free AA, which by itself did not stimulate superoxide generation, restored oxidase activity in neutrophils treated with PLA2 inhibitors. These findings indicate that phosphorylation and translocation of the cytosolic factors to the membranes are not sufficient for generating superoxide; a functional PLA2 is also needed to stimulate the oxidase activity. The inhibition of PLA2 activity did not prevent the phosphorylation of p47, suggesting that the location of PLA2 is downstream of and does not activate protein kinase C.
Full text
PDFImages in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Abo A., Pick E., Hall A., Totty N., Teahan C. G., Segal A. W. Activation of the NADPH oxidase involves the small GTP-binding protein p21rac1. Nature. 1991 Oct 17;353(6345):668–670. doi: 10.1038/353668a0. [DOI] [PubMed] [Google Scholar]
- Abo A., Pick E. Purification and characterization of a third cytosolic component of the superoxide-generating NADPH oxidase of macrophages. J Biol Chem. 1991 Dec 15;266(35):23577–23585. [PubMed] [Google Scholar]
- Abramson S. B., Leszczynska-Piziak J., Weissmann G. Arachidonic acid as a second messenger. Interactions with a GTP-binding protein of human neutrophils. J Immunol. 1991 Jul 1;147(1):231–236. [PubMed] [Google Scholar]
- Babior B. M. Protein phosphorylation and the respiratory burst. Arch Biochem Biophys. 1988 Aug 1;264(2):361–367. doi: 10.1016/0003-9861(88)90300-1. [DOI] [PubMed] [Google Scholar]
- Babior B. M. The respiratory burst of phagocytes. J Clin Invest. 1984 Mar;73(3):599–601. doi: 10.1172/JCI111249. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Badwey J. A., Robinson J. M., Heyworth P. G., Curnutte J. T. 1,2-dioctanoyl-sn-glycerol can stimulate neutrophils by different mechanisms. Evidence for a pathway that does not involve phosphorylation of the 47-kDa protein. J Biol Chem. 1989 Dec 5;264(34):20676–20682. [PubMed] [Google Scholar]
- Bromberg Y., Pick E. Unsaturated fatty acids as second messengers of superoxide generation by macrophages. Cell Immunol. 1983 Jul 15;79(2):240–252. doi: 10.1016/0008-8749(83)90067-9. [DOI] [PubMed] [Google Scholar]
- Clark J. D., Lin L. L., Kriz R. W., Ramesha C. S., Sultzman L. A., Lin A. Y., Milona N., Knopf J. L. A novel arachidonic acid-selective cytosolic PLA2 contains a Ca(2+)-dependent translocation domain with homology to PKC and GAP. Cell. 1991 Jun 14;65(6):1043–1051. doi: 10.1016/0092-8674(91)90556-e. [DOI] [PubMed] [Google Scholar]
- Combadière C., Hakim J., Giroud J. P., Périanin A. Staurosporine, a protein kinase inhibitor, up-regulates the stimulation of human neutrophil respiratory burst by N-formyl peptides and platelet activating factor. Biochem Biophys Res Commun. 1990 Apr 16;168(1):65–70. doi: 10.1016/0006-291x(90)91675-i. [DOI] [PubMed] [Google Scholar]
- Cross A. R., Jones O. T., Harper A. M., Segal A. W. Oxidation-reduction properties of the cytochrome b found in the plasma-membrane fraction of human neutrophils. A possible oxidase in the respiratory burst. Biochem J. 1981 Feb 15;194(2):599–606. doi: 10.1042/bj1940599. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dusi S., Della Bianca V., Grzeskowiak M., Rossi F. Relationship between phosphorylation and translocation to the plasma membrane of p47phox and p67phox and activation of the NADPH oxidase in normal and Ca(2+)-depleted human neutrophils. Biochem J. 1993 Feb 15;290(Pt 1):173–178. doi: 10.1042/bj2900173. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fujimori Y., Murakami M., Kim D. K., Hara S., Takayama K., Kudo I., Inoue K. Immunochemical detection of arachidonoyl-preferential phospholipase A2. J Biochem. 1992 Jan;111(1):54–60. doi: 10.1093/oxfordjournals.jbchem.a123718. [DOI] [PubMed] [Google Scholar]
- Gabig T. G., Lefker B. A. Catalytic properties of the resolved flavoprotein and cytochrome B components of the NADPH dependent O2- . generating oxidase from human neutrophils. Biochem Biophys Res Commun. 1984 Jan 30;118(2):430–436. doi: 10.1016/0006-291x(84)91321-4. [DOI] [PubMed] [Google Scholar]
- Garcia R. C., Segal A. W. Phosphorylation of the subunits of cytochrome b-245 upon triggering of the respiratory burst of human neutrophils and macrophages. Biochem J. 1988 Jun 15;252(3):901–904. doi: 10.1042/bj2520901. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gilliam E. B., Schulam P. G., Whelan J. P., Rosenblatt H. M., Shearer W. T. Phorbol ester plus calcium ionophore induces release of arachidonic acid from membrane phospholipids of a human B cell line. Cell Immunol. 1991 Aug;136(1):41–53. doi: 10.1016/0008-8749(91)90379-p. [DOI] [PubMed] [Google Scholar]
- Grinstein S., Furuya W. Receptor-mediated activation of electropermeabilized neutrophils. Evidence for a Ca2+- and protein kinase C-independent signaling pathway. J Biol Chem. 1988 Feb 5;263(4):1779–1783. [PubMed] [Google Scholar]
- Hayakawa T., Suzuki K., Suzuki S., Andrews P. C., Babior B. M. A possible role for protein phosphorylation in the activation of the respiratory burst in human neutrophils. Evidence from studies with cells from patients with chronic granulomatous disease. J Biol Chem. 1986 Jul 15;261(20):9109–9115. [PubMed] [Google Scholar]
- Henderson L. M., Chappell J. B., Jones O. T. Superoxide generation is inhibited by phospholipase A2 inhibitors. Role for phospholipase A2 in the activation of the NADPH oxidase. Biochem J. 1989 Nov 15;264(1):249–255. doi: 10.1042/bj2640249. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Heyworth P. G., Shrimpton C. F., Segal A. W. Localization of the 47 kDa phosphoprotein involved in the respiratory-burst NADPH oxidase of phagocytic cells. Biochem J. 1989 May 15;260(1):243–248. doi: 10.1042/bj2600243. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Huang K. P. The mechanism of protein kinase C activation. Trends Neurosci. 1989 Nov;12(11):425–432. doi: 10.1016/0166-2236(89)90091-x. [DOI] [PubMed] [Google Scholar]
- Khan W., el Touny S., Hannun Y. A. Arachidonic and cis-unsaturated fatty acids induce selective platelet substrate phosphorylation through activation of cytosolic protein kinase C. FEBS Lett. 1991 Nov 4;292(1-2):98–102. doi: 10.1016/0014-5793(91)80843-r. [DOI] [PubMed] [Google Scholar]
- Korchak H. M., Vienne K., Rutherford L. E., Weissmann G. Neutrophil stimulation: receptor, membrane, and metabolic events. Fed Proc. 1984 Sep;43(12):2749–2754. [PubMed] [Google Scholar]
- Kramer I. M., Verhoeven A. J., van der Bend R. L., Weening R. S., Roos D. Purified protein kinase C phosphorylates a 47-kDa protein in control neutrophil cytoplasts but not in neutrophil cytoplasts from patients with the autosomal form of chronic granulomatous disease. J Biol Chem. 1988 Feb 15;263(5):2352–2357. [PubMed] [Google Scholar]
- Leto T. L., Lomax K. J., Volpp B. D., Nunoi H., Sechler J. M., Nauseef W. M., Clark R. A., Gallin J. I., Malech H. L. Cloning of a 67-kD neutrophil oxidase factor with similarity to a noncatalytic region of p60c-src. Science. 1990 May 11;248(4956):727–730. doi: 10.1126/science.1692159. [DOI] [PubMed] [Google Scholar]
- Levy R., Rotrosen D., Nagauker O., Leto T. L., Malech H. L. Induction of the respiratory burst in HL-60 cells. Correlation of function and protein expression. J Immunol. 1990 Oct 15;145(8):2595–2601. [PubMed] [Google Scholar]
- Lin L. L., Lin A. Y., Knopf J. L. Cytosolic phospholipase A2 is coupled to hormonally regulated release of arachidonic acid. Proc Natl Acad Sci U S A. 1992 Jul 1;89(13):6147–6151. doi: 10.1073/pnas.89.13.6147. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lin L. L., Wartmann M., Lin A. Y., Knopf J. L., Seth A., Davis R. J. cPLA2 is phosphorylated and activated by MAP kinase. Cell. 1993 Jan 29;72(2):269–278. doi: 10.1016/0092-8674(93)90666-e. [DOI] [PubMed] [Google Scholar]
- Lomax K. J., Leto T. L., Nunoi H., Gallin J. I., Malech H. L. Recombinant 47-kilodalton cytosol factor restores NADPH oxidase in chronic granulomatous disease. Science. 1989 Jul 28;245(4916):409–412. doi: 10.1126/science.2547247. [DOI] [PubMed] [Google Scholar]
- Lu D. J., Takai A., Leto T. L., Grinstein S. Modulation of neutrophil activation by okadaic acid, a protein phosphatase inhibitor. Am J Physiol. 1992 Jan;262(1 Pt 1):C39–C49. doi: 10.1152/ajpcell.1992.262.1.C39. [DOI] [PubMed] [Google Scholar]
- MacEwan D. J., Mitchell R., Thomson F. J., Johnson M. S. Inhibition of depolarisation-induced calcium influx into GH3 cells by arachidonic acid: the involvement of protein kinase C. Biochim Biophys Acta. 1991 Sep 24;1094(3):346–354. doi: 10.1016/0167-4889(91)90096-g. [DOI] [PubMed] [Google Scholar]
- Majumdar S., Rossi M. W., Fujiki T., Phillips W. A., Disa S., Queen C. F., Johnston R. B., Jr, Rosen O. M., Corkey B. E., Korchak H. M. Protein kinase C isotypes and signaling in neutrophils. Differential substrate specificities of a translocatable calcium- and phospholipid-dependent beta-protein kinase C and a phospholipid-dependent protein kinase which is inhibited by long chain fatty acyl coenzyme A. J Biol Chem. 1991 May 15;266(14):9285–9294. [PubMed] [Google Scholar]
- Mayer R. J., Marshall L. A. New insights on mammalian phospholipase A2(s); comparison of arachidonoyl-selective and -nonselective enzymes. FASEB J. 1993 Feb 1;7(2):339–348. doi: 10.1096/fasebj.7.2.8440410. [DOI] [PubMed] [Google Scholar]
- McPhail L. C., Clayton C. C., Snyderman R. A potential second messenger role for unsaturated fatty acids: activation of Ca2+-dependent protein kinase. Science. 1984 May 11;224(4649):622–625. doi: 10.1126/science.6231726. [DOI] [PubMed] [Google Scholar]
- McPhail L. C., Clayton C. C., Snyderman R. The NADPH oxidase of human polymorphonuclear leukocytes. Evidence for regulation by multiple signals. J Biol Chem. 1984 May 10;259(9):5768–5775. [PubMed] [Google Scholar]
- Morel F., Doussiere J., Vignais P. V. The superoxide-generating oxidase of phagocytic cells. Physiological, molecular and pathological aspects. Eur J Biochem. 1991 Nov 1;201(3):523–546. doi: 10.1111/j.1432-1033.1991.tb16312.x. [DOI] [PubMed] [Google Scholar]
- Muid R. E., Twomey B., Dale M. M. The effect of inhibition of both diacylglycerol metabolism and phospholipase A2 activity on superoxide generation by human neutrophils. FEBS Lett. 1988 Jul 4;234(1):235–240. doi: 10.1016/0014-5793(88)81342-5. [DOI] [PubMed] [Google Scholar]
- Müller S., Nigam S. Arachidonic acid release and platelet-activating factor formation by staurosporine in human neutrophils challenged with n-formyl peptide. Eur J Pharmacol. 1992 Aug 6;218(2-3):251–258. doi: 10.1016/0014-2999(92)90176-5. [DOI] [PubMed] [Google Scholar]
- Nakashima S., Suganuma A., Sato M., Tohmatsu T., Nozawa Y. Mechanism of arachidonic acid liberation in platelet-activating factor-stimulated human polymorphonuclear neutrophils. J Immunol. 1989 Aug 15;143(4):1295–1302. [PubMed] [Google Scholar]
- Naor Z. Is arachidonic acid a second messenger in signal transduction? Mol Cell Endocrinol. 1991 Sep;80(1-3):C181–C186. doi: 10.1016/0303-7207(91)90135-f. [DOI] [PubMed] [Google Scholar]
- Naor Z., Shearman M. S., Kishimoto A., Nishizuka Y. Calcium-independent activation of hypothalamic type I protein kinase C by unsaturated fatty acids. Mol Endocrinol. 1988 Nov;2(11):1043–1048. doi: 10.1210/mend-2-11-1043. [DOI] [PubMed] [Google Scholar]
- Nishizuka Y. The molecular heterogeneity of protein kinase C and its implications for cellular regulation. Nature. 1988 Aug 25;334(6184):661–665. doi: 10.1038/334661a0. [DOI] [PubMed] [Google Scholar]
- Okamura N., Curnutte J. T., Roberts R. L., Babior B. M. Relationship of protein phosphorylation to the activation of the respiratory burst in human neutrophils. Defects in the phosphorylation of a group of closely related 48-kDa proteins in two forms of chronic granulomatous disease. J Biol Chem. 1988 May 15;263(14):6777–6782. [PubMed] [Google Scholar]
- Parker J., Daniel L. W., Waite M. Evidence of protein kinase C involvement in phorbol diester-stimulated arachidonic acid release and prostaglandin synthesis. J Biol Chem. 1987 Apr 15;262(11):5385–5393. [PubMed] [Google Scholar]
- Pernas P., Masliah J., Olivier J. L., Salvat C., Rybkine T., Bereziat G. Type II phospholipase A2 recombinant overexpression enhances stimulated arachidonic acid release. Biochem Biophys Res Commun. 1991 Aug 15;178(3):1298–1305. doi: 10.1016/0006-291x(91)91035-b. [DOI] [PubMed] [Google Scholar]
- Reibman J., Korchak H. M., Vosshall L. B., Haines K. A., Rich A. M., Weissmann G. Changes in diacylglycerol labeling, cell shape, and protein phosphorylation distinguish "triggering" from "activation" of human neutrophils. J Biol Chem. 1988 May 5;263(13):6322–6328. [PubMed] [Google Scholar]
- Rossi F. The O2- -forming NADPH oxidase of the phagocytes: nature, mechanisms of activation and function. Biochim Biophys Acta. 1986 Nov 4;853(1):65–89. doi: 10.1016/0304-4173(86)90005-4. [DOI] [PubMed] [Google Scholar]
- Rotrosen D., Leto T. L. Phosphorylation of neutrophil 47-kDa cytosolic oxidase factor. Translocation to membrane is associated with distinct phosphorylation events. J Biol Chem. 1990 Nov 15;265(32):19910–19915. [PubMed] [Google Scholar]
- Rotrosen D., Yeung C. L., Leto T. L., Malech H. L., Kwong C. H. Cytochrome b558: the flavin-binding component of the phagocyte NADPH oxidase. Science. 1992 Jun 5;256(5062):1459–1462. doi: 10.1126/science.1318579. [DOI] [PubMed] [Google Scholar]
- Rubinek T., Levy R. Arachidonic acid increases the activity of the assembled NADPH oxidase in cytoplasmic membranes and endosomes. Biochim Biophys Acta. 1993 Mar 10;1176(1-2):51–58. doi: 10.1016/0167-4889(93)90176-p. [DOI] [PubMed] [Google Scholar]
- Sakata A., Ida E., Tominaga M., Onoue K. Arachidonic acid acts as an intracellular activator of NADPH-oxidase in Fc gamma receptor-mediated superoxide generation in macrophages. J Immunol. 1987 Jun 15;138(12):4353–4359. [PubMed] [Google Scholar]
- Segal A. W., Heyworth P. G., Cockcroft S., Barrowman M. M. Stimulated neutrophils from patients with autosomal recessive chronic granulomatous disease fail to phosphorylate a Mr-44,000 protein. Nature. 1985 Aug 8;316(6028):547–549. doi: 10.1038/316547a0. [DOI] [PubMed] [Google Scholar]
- Segal A. W., West I., Wientjes F., Nugent J. H., Chavan A. J., Haley B., Garcia R. C., Rosen H., Scrace G. Cytochrome b-245 is a flavocytochrome containing FAD and the NADPH-binding site of the microbicidal oxidase of phagocytes. Biochem J. 1992 Jun 15;284(Pt 3):781–788. doi: 10.1042/bj2840781. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sekiguchi K., Tsukuda M., Ogita K., Kikkawa U., Nishizuka Y. Three distinct forms of rat brain protein kinase C: differential response to unsaturated fatty acids. Biochem Biophys Res Commun. 1987 Jun 15;145(2):797–802. doi: 10.1016/0006-291x(87)91035-7. [DOI] [PubMed] [Google Scholar]
- Sharp J. D., White D. L., Chiou X. G., Goodson T., Gamboa G. C., McClure D., Burgett S., Hoskins J., Skatrud P. L., Sportsman J. R. Molecular cloning and expression of human Ca(2+)-sensitive cytosolic phospholipase A2. J Biol Chem. 1991 Aug 15;266(23):14850–14853. [PubMed] [Google Scholar]
- Smolen J. E., Weissmann G. Effects of indomethacin, 5,8,11,14-eicosatetraynoic acid, and p-bromophenacyl bromide on lysosomal enzyme release and superoxide anion generation by human polymorphonuclear leukocytes. Biochem Pharmacol. 1980 Feb 15;29(4):533–538. doi: 10.1016/0006-2952(80)90373-1. [DOI] [PubMed] [Google Scholar]
- Svensson U., Holst E., Sundler R. Protein-kinase-C-independent activation of arachidonate release and prostaglandin E2 formation in macrophages interacting with certain bacteria. Eur J Biochem. 1991 Sep 15;200(3):699–705. doi: 10.1111/j.1432-1033.1991.tb16234.x. [DOI] [PubMed] [Google Scholar]
- Tarsi-Tsuk D., Levy R. Stimulation of the respiratory burst in peripheral blood monocytes by lipoteichoic acid. The involvement of calcium ions and phospholipase A2. J Immunol. 1990 Apr 1;144(7):2665–2670. [PubMed] [Google Scholar]
- Volpp B. D., Nauseef W. M., Donelson J. E., Moser D. R., Clark R. A. Cloning of the cDNA and functional expression of the 47-kilodalton cytosolic component of human neutrophil respiratory burst oxidase. Proc Natl Acad Sci U S A. 1989 Sep;86(18):7195–7199. doi: 10.1073/pnas.86.18.7195. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zor U., Her E., Harell T., Fischer G., Naor Z., Braquet P., Ferber E., Reiss N. Arachidonic acid release by basophilic leukemia cells and macrophages stimulated by Ca2+ ionophores, antigen and diacylglycerol: essential role for protein kinase C and prevention by glucocorticosteroids. Biochim Biophys Acta. 1991 Feb 19;1091(3):385–392. doi: 10.1016/0167-4889(91)90204-b. [DOI] [PubMed] [Google Scholar]