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. 1991 Dec 1;280(Pt 2):379–385. doi: 10.1042/bj2800379

Studies on the activation of human neutrophil 5-lipoxygenase induced by natural agonists and Ca2+ ionophore A23187.

P P McDonald 1, S R McColl 1, P H Naccache 1, P Borgeat 1
PMCID: PMC1130557  PMID: 1747110

Abstract

By using exogenous substrates, activation of human neutrophil 5-lipoxygenase can be investigated independently of the release of endogenous arachidonic acid. We have developed a sensitive assay to measure 5-LO activation which takes advantage of the 5-LO-mediated conversion of 15S-hydroperoxy-5,8,11,13(Z,Z,Z,E)-eicosatetraenoic acid (15-HpETE) into 5S,15S-dihydroxy-6,8,11,13(E,Z,Z,E)-eicosatetraenoic acid (5,15-DiHETE). When resting neutrophils were incubated with low micromolar concentrations of 15-HpETE, a minor dose- and time-dependent formation of 5,15-DiHETE was observed. In contrast, co-addition of 15-HpETE with Ca2+ ionophore A23187 or with the neutrophil agonists platelet-activating factor (PAF), fMetLeuPhe or complement component C5a resulted in a sizeable concentration-dependent synthesis of 5,15-DiHETE, while lyso-PAF and phorbol myristate acetate were without effect on 5,15-DiHETE formation from 15-HpETE. This stimulation of 5,15-DiHETE synthesis by A23187 or by natural agonists was effectively inhibited by MK-886, a compound that has recently been reported to inhibit the A23187-induced translocation of 5-LO to membrane structures. Furthermore, natural-agonist-induced activation of the 5-LO-mediated transformation of 15-HpETE was inhibited by pertussis toxin, indicating the involvement of a GTP-binding protein in the 5-LO activation process.

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Selected References

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  1. Borgeat P., Fruteau de Laclos B., Maclouf J. New concepts in the modulation of leukotriene synthesis. Biochem Pharmacol. 1983 Feb 1;32(3):381–387. doi: 10.1016/0006-2952(83)90515-4. [DOI] [PubMed] [Google Scholar]
  2. Borgeat P., Picard S. 19-Hydroxyprostaglandin B2 as an internal standard for on-line extraction-high-performance liquid chromatography analysis of lipoxygenase products. Anal Biochem. 1988 Jun;171(2):283–289. doi: 10.1016/0003-2697(88)90487-3. [DOI] [PubMed] [Google Scholar]
  3. Borgeat P., Samuelsson B. Arachidonic acid metabolism in polymorphonuclear leukocytes: effects of ionophore A23187. Proc Natl Acad Sci U S A. 1979 May;76(5):2148–2152. doi: 10.1073/pnas.76.5.2148. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bradford P. G., Rubin R. P. Characterization of formylmethionyl-leucyl-phenylalanine stimulation of inositol trisphosphate accumulation in rabbit neutrophils. Mol Pharmacol. 1985 Jan;27(1):74–78. [PubMed] [Google Scholar]
  5. Chenoweth D. E., Hugli T. E. Demonstration of specific C5a receptor on intact human polymorphonuclear leukocytes. Proc Natl Acad Sci U S A. 1978 Aug;75(8):3943–3947. doi: 10.1073/pnas.75.8.3943. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chilton F. H., O'Flaherty J. T., Walsh C. E., Thomas M. J., Wykle R. L., DeChatelet L. R., Waite B. M. Platelet activating factor. Stimulation of the lipoxygenase pathway in polymorphonuclear leukocytes by 1-O-alkyl-2-O-acetyl-sn-glycero-3-phosphocholine. J Biol Chem. 1982 May 25;257(10):5402–5407. [PubMed] [Google Scholar]
  7. Claesson H. E., Lundberg U., Malmsten C. Serum-coated zymosan stimulates the synthesis of leukotriene B4 in human polymorphonuclear leukocytes. Inhibition by cyclic AMP. Biochem Biophys Res Commun. 1981 Apr 30;99(4):1230–1237. doi: 10.1016/0006-291x(81)90751-8. [DOI] [PubMed] [Google Scholar]
  8. Clancy R. M., Dahinden C. A., Hugli T. E. Arachidonate metabolism by human polymorphonuclear leukocytes stimulated by N-formyl-Met-Leu-Phe or complement component C5a is independent of phospholipase activation. Proc Natl Acad Sci U S A. 1983 Dec;80(23):7200–7204. doi: 10.1073/pnas.80.23.7200. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dixon R. A., Diehl R. E., Opas E., Rands E., Vickers P. J., Evans J. F., Gillard J. W., Miller D. K. Requirement of a 5-lipoxygenase-activating protein for leukotriene synthesis. Nature. 1990 Jan 18;343(6255):282–284. doi: 10.1038/343282a0. [DOI] [PubMed] [Google Scholar]
  10. Ford-Hutchinson A. W., Bray M. A., Doig M. V., Shipley M. E., Smith M. J. Leukotriene B, a potent chemokinetic and aggregating substance released from polymorphonuclear leukocytes. Nature. 1980 Jul 17;286(5770):264–265. doi: 10.1038/286264a0. [DOI] [PubMed] [Google Scholar]
  11. Hafstrom I., Palmblad J., Malmsten C. L., Rådmark O., Samuelsson B. Leukotriene B4--a stereospecific stimulator for release of lysosomal enzymes from neutrophils. FEBS Lett. 1981 Jul 20;130(1):146–148. doi: 10.1016/0014-5793(81)80684-9. [DOI] [PubMed] [Google Scholar]
  12. Hatzelmann A., Schatz M., Ullrich V. Involvement of glutathione peroxidase activity in the stimulation of 5-lipoxygenase activity by glutathione-depleting agents in human polymorphonuclear leukocytes. Eur J Biochem. 1989 Apr 1;180(3):527–533. doi: 10.1111/j.1432-1033.1989.tb14678.x. [DOI] [PubMed] [Google Scholar]
  13. Hatzelmann A., Ullrich V. Regulation of 5-lipoxygenase activity by the glutathione status in human polymorphonuclear leukocytes. Eur J Biochem. 1987 Nov 16;169(1):175–184. doi: 10.1111/j.1432-1033.1987.tb13595.x. [DOI] [PubMed] [Google Scholar]
  14. Haurand M., Flohé L. Leukotriene formation by human polymorphonuclear leukocytes from endogenous arachidonate. Physiological triggers and modulation by prostanoids. Biochem Pharmacol. 1989 Jul 1;38(13):2129–2137. doi: 10.1016/0006-2952(89)90067-1. [DOI] [PubMed] [Google Scholar]
  15. Lad P. M., Olson C. V., Grewal I. S. Platelet-activating factor mediated effects on human neutrophil function are inhibited by pertussis toxin. Biochem Biophys Res Commun. 1985 Jun 28;129(3):632–638. doi: 10.1016/0006-291x(85)91938-2. [DOI] [PubMed] [Google Scholar]
  16. McColl S. R., Krump E., Naccache P. H., Caon A. C., Borgeat P. Activation of the human neutrophil 5-lipoxygenase by exogenous arachidonic acid: involvement of pertussis toxin-sensitive guanine nucleotide-binding proteins. Br J Pharmacol. 1989 Aug;97(4):1265–1273. doi: 10.1111/j.1476-5381.1989.tb12588.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Miller D. K., Gillard J. W., Vickers P. J., Sadowski S., Léveillé C., Mancini J. A., Charleson P., Dixon R. A., Ford-Hutchinson A. W., Fortin R. Identification and isolation of a membrane protein necessary for leukotriene production. Nature. 1990 Jan 18;343(6255):278–281. doi: 10.1038/343278a0. [DOI] [PubMed] [Google Scholar]
  18. Molski T. F., Naccache P. H., Marsh M. L., Kermode J., Becker E. L., Sha'afi R. I. Pertussis toxin inhibits the rise in the intracellular concentration of free calcium that is induced by chemotactic factors in rabbit neutrophils: possible role of the "G proteins" in calcium mobilization. Biochem Biophys Res Commun. 1984 Oct 30;124(2):644–650. doi: 10.1016/0006-291x(84)91603-6. [DOI] [PubMed] [Google Scholar]
  19. Ménard L., Pilote S., Naccache P. H., Laviolette M., Borgeat P. Inhibitory effects of MK-886 on arachidonic acid metabolism in human phagocytes. Br J Pharmacol. 1990 May;100(1):15–20. doi: 10.1111/j.1476-5381.1990.tb12044.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Naccache P. H., Faucher N., Caon A. C., McColl S. R. Propionic acid-induced calcium mobilization in human neutrophils. J Cell Physiol. 1988 Jul;136(1):118–124. doi: 10.1002/jcp.1041360115. [DOI] [PubMed] [Google Scholar]
  21. Naccache P. H., McColl S. R., Caon A. C., Borgeat P. Arachidonic acid-induced mobilization of calcium in human neutrophils: evidence for a multicomponent mechanism of action. Br J Pharmacol. 1989 Jun;97(2):461–468. doi: 10.1111/j.1476-5381.1989.tb11973.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Naccache P. H., Molski M. M., Volpi M., Becker E. L., Sha'afi R. I. Unique inhibitory profile of platelet activating factor induced calcium mobilization, polyphosphoinositide turnover and granule enzyme secretion in rabbit neutrophils towards pertussis toxin and phorbol ester. Biochem Biophys Res Commun. 1985 Jul 31;130(2):677–684. doi: 10.1016/0006-291x(85)90470-x. [DOI] [PubMed] [Google Scholar]
  23. Naccache P. H., Sha'afi R. I., Borgeat P., Goetzl E. J. Mono- and dihydroxyeicosatetraenoic acids alter calcium homeostasis in rabbit neutrophils. J Clin Invest. 1981 May;67(5):1584–1587. doi: 10.1172/JCI110191. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Naccache P. H., Volpi M., Showell H. J., Becker E. L., Sha'afi R. I. Chemotactic factor-induced release of membrane calcium in rabbit neutrophils. Science. 1979 Feb 2;203(4379):461–463. doi: 10.1126/science.760200. [DOI] [PubMed] [Google Scholar]
  25. Nagy L., Lee T. H., Goetzl E. J., Pickett W. C., Kay A. B. Complement receptor enhancement and chemotaxis of human neutrophils and eosinophils by leukotrienes and other lipoxygenase products. Clin Exp Immunol. 1982 Mar;47(3):541–547. [PMC free article] [PubMed] [Google Scholar]
  26. Nichols R. C., Vanderhoek J. Y. 5-Hydroxyeicosanoids selectively stimulate the human neutrophil 15-lipoxygenase to use endogenous substrate. J Exp Med. 1990 Feb 1;171(2):367–375. doi: 10.1084/jem.171.2.367. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Palmblad J., Malmsten C. L., Udén A. M., Rådmark O., Engstedt L., Samuelsson B. Leukotriene B4 is a potent and stereospecific stimulator of neutrophil chemotaxis and adherence. Blood. 1981 Sep;58(3):658–661. [PubMed] [Google Scholar]
  28. Poubelle P. E., De Medicis R., Naccache P. H. Monosodium urate and calcium pyrophosphate crystals differentially activate the excitation-response coupling sequence of human neutrophils. Biochem Biophys Res Commun. 1987 Dec 16;149(2):649–657. doi: 10.1016/0006-291x(87)90417-7. [DOI] [PubMed] [Google Scholar]
  29. Rola-Pleszczynski M., Lemaire I. Leukotrienes augment interleukin 1 production by human monocytes. J Immunol. 1985 Dec;135(6):3958–3961. [PubMed] [Google Scholar]
  30. Rouzer C. A., Ford-Hutchinson A. W., Morton H. E., Gillard J. W. MK886, a potent and specific leukotriene biosynthesis inhibitor blocks and reverses the membrane association of 5-lipoxygenase in ionophore-challenged leukocytes. J Biol Chem. 1990 Jan 25;265(3):1436–1442. [PubMed] [Google Scholar]
  31. Rouzer C. A., Kargman S. Translocation of 5-lipoxygenase to the membrane in human leukocytes challenged with ionophore A23187. J Biol Chem. 1988 Aug 5;263(22):10980–10988. [PubMed] [Google Scholar]
  32. Rouzer C. A., Rands E., Kargman S., Jones R. E., Register R. B., Dixon R. A. Characterization of cloned human leukocyte 5-lipoxygenase expressed in mammalian cells. J Biol Chem. 1988 Jul 25;263(21):10135–10140. [PubMed] [Google Scholar]
  33. Salari H., Braquet P., Naccache P., Borgeat P. Characterization of effect of N-formyl-methionyl-leucyl-phenylalanine on leukotriene synthesis in human polymorphonuclear leukocytes. Inflammation. 1985 Jun;9(2):127–138. doi: 10.1007/BF00917585. [DOI] [PubMed] [Google Scholar]
  34. Schröder J. M. The monocyte-derived neutrophil activating peptide (NAP/interleukin 8) stimulates human neutrophil arachidonate-5-lipoxygenase, but not the release of cellular arachidonate. J Exp Med. 1989 Sep 1;170(3):847–863. doi: 10.1084/jem.170.3.847. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Shirato M., Takahashi K., Nagasawa S., Koyama J. Different sensitivities of the responses of human neutrophils stimulated with immune complex and C5a anaphylatoxin to pertussis toxin. FEBS Lett. 1988 Jul 4;234(1):231–234. doi: 10.1016/0014-5793(88)81341-3. [DOI] [PubMed] [Google Scholar]
  36. Tsien R. Y. A non-disruptive technique for loading calcium buffers and indicators into cells. Nature. 1981 Apr 9;290(5806):527–528. doi: 10.1038/290527a0. [DOI] [PubMed] [Google Scholar]
  37. Verghese M. W., Charles L., Jakoi L., Dillon S. B., Snyderman R. Role of a guanine nucleotide regulatory protein in the activation of phospholipase C by different chemoattractants. J Immunol. 1987 Jun 15;138(12):4374–4380. [PubMed] [Google Scholar]
  38. White J. R., Naccache P. H., Molski T. F., Borgeat P., Sha'afi R. I. Direct demonstration of increased intracellular concentration of free calcium in rabbit and human neutrophils following stimulation by chemotactic factor. Biochem Biophys Res Commun. 1983 May 31;113(1):44–50. doi: 10.1016/0006-291x(83)90429-1. [DOI] [PubMed] [Google Scholar]
  39. Wong A., Hwang S. M., Cook M. N., Hogaboom G. K., Crooke S. T. Interactions of 5-lipoxygenase with membranes: studies on the association of soluble enzyme with membranes and alterations in enzyme activity. Biochemistry. 1988 Sep 6;27(18):6763–6769. doi: 10.1021/bi00418a018. [DOI] [PubMed] [Google Scholar]

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