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. 1985 Sep 1;101(3):1052–1058. doi: 10.1083/jcb.101.3.1052

Release of superoxide and change in morphology by neutrophils in response to phorbol esters: antagonism by inhibitors of calcium-binding proteins

PMCID: PMC2113709  PMID: 2993312

Abstract

The ability of phorbol derivatives to function as stimulating agents for superoxide (O2-) release by guinea pig neutrophils has been evaluated and compared to the known ability of each compound to activate protein kinase C. Those that activate the kinase also stimulate O2- release, while those that are inactive with respect to the kinase have no effect on O2- release. The same correlation was observed with respect to the ability of phorbol esters to induce morphological changes in neutrophils, i.e., vesiculation and reduction in granule content. Certain phenothiazines and naphthalene sulfonamides that are known antagonists of calcium-binding proteins blocked both phorbol ester-induced O2- release and morphological changes in these cells.

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Selected References

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  1. Alobaidi T., Naccache P. H., Sha'afi R. I. Calmodulin antagonists modulate rabbit neutrophil degranulation, aggregation and stimulated oxygen consumption. Biochim Biophys Acta. 1981 Jul 17;675(3-4):316–321. doi: 10.1016/0304-4165(81)90020-9. [DOI] [PubMed] [Google Scholar]
  2. Andrews P. C., Babior B. M. Phosphorylation of cytosolic proteins by resting and activated human neutrophils. Blood. 1984 Oct;64(4):883–890. [PubMed] [Google Scholar]
  3. 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]
  4. Badwey J. A., Curnutte J. T., Robinson J. M., Berde C. B., Karnovsky M. J., Karnovsky M. L. Effects of free fatty acids on release of superoxide and on change of shape by human neutrophils. Reversibility by albumin. J Biol Chem. 1984 Jun 25;259(12):7870–7877. [PubMed] [Google Scholar]
  5. Badwey J. A., Curnutte J. T., Robinson J. M., Lazdins J. K., Briggs R. T., Karnovsky M. J., Karnovsky M. L. Comparative aspects of oxidative metabolism of neutrophils from human blood and guinea pig peritonea: magnitude of the respiratory burst, dependence upon stimulating agents, and localization of the oxidases. J Cell Physiol. 1980 Dec;105(3):541–545. doi: 10.1002/jcp.1041050319. [DOI] [PubMed] [Google Scholar]
  6. Badwey J. A., Karnovsky M. L. Active oxygen species and the functions of phagocytic leukocytes. Annu Rev Biochem. 1980;49:695–726. doi: 10.1146/annurev.bi.49.070180.003403. [DOI] [PubMed] [Google Scholar]
  7. Badwey J. A., Robinson J. M., Lazdins J. K., Briggs R. T., Karnovsky M. J., Karnovsky M. L. Comparative biochemical and cytochemical studies on superoxide and peroxide in mouse macrophages. J Cell Physiol. 1983 May;115(2):208–216. doi: 10.1002/jcp.1041150216. [DOI] [PubMed] [Google Scholar]
  8. Becker E. L., Sigman M., Oliver J. M. Superoxide production induced in rabbit polymorphonuclear leukocytes by synthetic chemotactic peptides and A23187. Am J Pathol. 1979 Apr;95(1):81–97. [PMC free article] [PubMed] [Google Scholar]
  9. Castagna M., Takai Y., Kaibuchi K., Sano K., Kikkawa U., Nishizuka Y. Direct activation of calcium-activated, phospholipid-dependent protein kinase by tumor-promoting phorbol esters. J Biol Chem. 1982 Jul 10;257(13):7847–7851. [PubMed] [Google Scholar]
  10. Cheung W. Y. Calmodulin plays a pivotal role in cellular regulation. Science. 1980 Jan 4;207(4426):19–27. doi: 10.1126/science.6243188. [DOI] [PubMed] [Google Scholar]
  11. Cohen H. J., Chovaniec M. E., Ellis S. E. Chlorpromazine inhibition of granulocyte superoxide production. Blood. 1980 Jul;56(1):23–29. [PubMed] [Google Scholar]
  12. Cohen H. J., Chovaniec M. E. Superoxide generation by digitonin-stimulated guinea pig granulocytes. A basis for a continuous assay for monitoring superoxide production and for the study of the activation of the generating system. J Clin Invest. 1978 Apr;61(4):1081–1087. doi: 10.1172/JCI109007. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Couturier A., Bazgar S., Castagna M. Further characterization of tumor-promoter-mediated activation of protein kinase C. Biochem Biophys Res Commun. 1984 Jun 15;121(2):448–455. doi: 10.1016/0006-291x(84)90203-1. [DOI] [PubMed] [Google Scholar]
  14. Curnutte J. T., Babior B. M. Biological defense mechanisms. The effect of bacteria and serum on superoxide production by granulocytes. J Clin Invest. 1974 Jun;53(6):1662–1672. doi: 10.1172/JCI107717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Curnutte J. T., Badwey J. A., Robinson J. M., Karnovsky M. J., Karnovsky M. L. Studies on the mechanism of superoxide release from human neutrophils stimulated with arachidonate. J Biol Chem. 1984 Oct 10;259(19):11851–11857. [PubMed] [Google Scholar]
  16. DULBECCO R., VOGT M. Plaque formation and isolation of pure lines with poliomyelitis viruses. J Exp Med. 1954 Feb;99(2):167–182. doi: 10.1084/jem.99.2.167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Davis B. H., Walter R. J., Pearson C. B., Becker E. L., Oliver J. M. Membrane activity and topography of F-Met-Leu-Phe-Treated polymorphonuclear leukocytes. Acute and sustained responses to chemotactic peptide. Am J Pathol. 1982 Aug;108(2):206–216. [PMC free article] [PubMed] [Google Scholar]
  18. DePierre J. W., Karnovsky M. L. Ecto-enzymes of the guinea pig polymorphonuclear leukocyte. I. Evidence for an ecto-adenosine monophosphatase, adenosine triphosphatase, and -p-nitrophenyl phosphates. J Biol Chem. 1974 Nov 25;249(22):7111–7120. [PubMed] [Google Scholar]
  19. Fujita I., Irita K., Takeshige K., Minakami S. Diacylglycerol, 1-oleoyl-2-acetyl-glycerol, stimulates superoxide-generation from human neutrophils. Biochem Biophys Res Commun. 1984 Apr 30;120(2):318–324. doi: 10.1016/0006-291x(84)91256-7. [DOI] [PubMed] [Google Scholar]
  20. Goodwin B. J., Weinberg J. B. Receptor-mediated modulation of human monocyte, neutrophil, lymphocyte, and platelet function by phorbol diesters. J Clin Invest. 1982 Oct;70(4):699–706. doi: 10.1172/JCI110665. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hidaka H., Sasaki Y., Tanaka T., Endo T., Ohno S., Fujii Y., Nagata T. N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide, a calmodulin antagonist, inhibits cell proliferation. Proc Natl Acad Sci U S A. 1981 Jul;78(7):4354–4357. doi: 10.1073/pnas.78.7.4354. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Hoffstein S. T., Friedman R. S., Weissmann G. Degranulation, membrane addition, and shape change during chemotactic factor-induced aggregation of human neutrophils. J Cell Biol. 1982 Oct;95(1):234–241. doi: 10.1083/jcb.95.1.234. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Johnston R. B., Jr, Lehmeyer J. E. Elaboration of toxic oxygen by-products by neutrophils in a model of immune complex disease. J Clin Invest. 1976 Apr;57(4):836–841. doi: 10.1172/JCI108359. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Jones H. P., Ghai G., Petrone W. F., McCord J. M. Calmodulin-dependent stimulation of the NADPH oxidase of human neutrophils. Biochim Biophys Acta. 1982 Jan 12;714(1):152–156. doi: 10.1016/0304-4165(82)90137-4. [DOI] [PubMed] [Google Scholar]
  25. Kishimoto A., Takai Y., Mori T., Kikkawa U., Nishizuka Y. Activation of calcium and phospholipid-dependent protein kinase by diacylglycerol, its possible relation to phosphatidylinositol turnover. J Biol Chem. 1980 Mar 25;255(6):2273–2276. [PubMed] [Google Scholar]
  26. Kuo J. F., Schatzman R. C., Turner R. S., Mazzei G. J. Phospholipid-sensitive Ca2+-dependent protein kinase: a major protein phosphorylation system. Mol Cell Endocrinol. 1984 May;35(2-3):65–73. doi: 10.1016/0303-7207(84)90001-7. [DOI] [PubMed] [Google Scholar]
  27. Lehrer R. I., Cohen L. Receptor-mediated regulation of superoxide production in human neutrophils stimulated by phorbol myristate acetate. J Clin Invest. 1981 Nov;68(5):1314–1320. doi: 10.1172/JCI110378. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Maupin P., Pollard T. D. Improved preservation and staining of HeLa cell actin filaments, clathrin-coated membranes, and other cytoplasmic structures by tannic acid-glutaraldehyde-saponin fixation. J Cell Biol. 1983 Jan;96(1):51–62. doi: 10.1083/jcb.96.1.51. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. 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]
  30. Mori T., Takai Y., Minakuchi R., Yu B., Nishizuka Y. Inhibitory action of chlorpromazine, dibucaine, and other phospholipid-interacting drugs on calcium-activated, phospholipid-dependent protein kinase. J Biol Chem. 1980 Sep 25;255(18):8378–8380. [PubMed] [Google Scholar]
  31. Naccache P. H., Molski T. F., Alobaidi T., Becker E. L., Showell H. J., Sha'afi R. I. Calmodulin inhibitors block neutrophil degranulation at a step distal from the mobilization of calcium. Biochem Biophys Res Commun. 1980 Nov 17;97(1):62–68. doi: 10.1016/s0006-291x(80)80134-3. [DOI] [PubMed] [Google Scholar]
  32. Nelson G. A., Andrews M. L., Karnovsky M. J. Control of erythrocyte shape by calmodulin. J Cell Biol. 1983 Mar;96(3):730–735. doi: 10.1083/jcb.96.3.730. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Niedel J. E., Kuhn L. J., Vandenbark G. R. Phorbol diester receptor copurifies with protein kinase C. Proc Natl Acad Sci U S A. 1983 Jan;80(1):36–40. doi: 10.1073/pnas.80.1.36. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Repine J. E., White J. G., Clawson C. C., Holmes B. M. Effects of phorbol myristate acetate on the metabolism and ultrastructure of neutrophils in chronic granulomatous disease. J Clin Invest. 1974 Jul;54(1):83–90. doi: 10.1172/JCI107752. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Robinson J. M., Badwey J. A., Karnovsky M. L., Karnovsky M. J. Superoxide release by neutrophils: synergistic effects of a phorbol ester and a calcium ionophore. Biochem Biophys Res Commun. 1984 Jul 31;122(2):734–739. doi: 10.1016/s0006-291x(84)80095-9. [DOI] [PubMed] [Google Scholar]
  36. Robinson J. M., Karnovsky M. L., Karnovsky M. J. Glycogen accumulation in polymorphonuclear leukocytes, and other intracellular alterations that occur during inflammation. J Cell Biol. 1982 Dec;95(3):933–942. doi: 10.1083/jcb.95.3.933. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Smith R. J., Bowman B. J., Iden S. S. Effects of trifluoperazine on human neutrophil function. Immunology. 1981 Dec;44(4):677–684. [PMC free article] [PubMed] [Google Scholar]
  38. Smolen J. E., Korchak H. M., Weissmann G. The roles of extracellular and intracellular calcium in lysosomal enzyme release and superoxide anion generation by human neutrophils. Biochim Biophys Acta. 1981 Nov 5;677(3-4):512–520. doi: 10.1016/0304-4165(81)90267-1. [DOI] [PubMed] [Google Scholar]
  39. TAKETA K., POGELL B. M. ALLOSTERIC INHIBITION OF RAT LIVER FRUCTOSE 1,6-DIPHOSPHATASE BY ADENOSINE 5'-MONOPHOSPHATE. J Biol Chem. 1965 Feb;240:651–662. [PubMed] [Google Scholar]
  40. Tauber A. I., Brettler D. B., Kennington E. A., Blumberg P. M. Relation of human neutrophil phorbol ester receptor occupancy and NADPH-oxidase activity. Blood. 1982 Aug;60(2):333–339. [PubMed] [Google Scholar]
  41. Vandenbark G. R., Kuhn L. J., Niedel J. E. Possible mechanism of phorbol diester-induced maturation of human promyelocytic leukemia cells. J Clin Invest. 1984 Feb;73(2):448–457. doi: 10.1172/JCI111231. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Weiss B., Prozialeck W., Cimino M., Barnette M. S., Wallace T. L. Pharmacological regulation of calmodulin. Ann N Y Acad Sci. 1980;356:319–345. doi: 10.1111/j.1749-6632.1980.tb29621.x. [DOI] [PubMed] [Google Scholar]
  43. White J. R., Huang C. K., Hill J. M., Jr, Naccache P. H., Becker E. L., Sha'afi R. I. Effect of phorbol 12-myristate 13-acetate and its analogue 4 alpha-phorbol 12,13-didecanoate on protein phosphorylation and lysosomal enzyme release in rabbit neutrophils. J Biol Chem. 1984 Jul 10;259(13):8605–8611. [PubMed] [Google Scholar]

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