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. 1984 Mar;51(3):571–583.

Subcellular localization of enzymes involved in leukotriene formation within human polymorphonuclear granulocytes.

J Brom, M Raulf, M Stüning, B Spur, A Crea, K D Bremm, W König
PMCID: PMC1454457  PMID: 6141997

Abstract

The subcellular localization of enzymes involved in leukotriene formation was analysed according to biological (chemotaxis, spasmogenic properties) and analytical methods. By subcellular fractionation the major activity for 5--lipoxygenase and L-gamma-glutamyltranspeptidase coeluted with the 200,000 g precipitate while glutathione-S-transferase activity was mainly present in the 200,000 g supernatant. Our data were supported by results indicating that the 200,000 g precipitate and supernatant fractions proved to be most active in generating 5-HETE and leukotriene C4 (LTC4) respectively. The 200,000 g pellet was the most active fraction in transforming synthetic LTC4 into LTD4 and LTE4. When synthetic LTD4 was incubated with the various subcellular fractions and the appearance of LTE4 was analysed the 20,000 and 200,000 g pellets were the most potent fractions. Several discrepancies observed using biological, biochemical and analytical (synthetic substrates) methods may be in part due to the formation of leukotriene isomers which interfere with the biological assays.

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Selected References

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  1. Aehringhaus U., Wölbling R. H., König W., Patrono C., Peskar B. M., Peskar B. A. Release of leukotriene C4 from human polymorphonuclear leucocytes as determined by radioimmunoassay. FEBS Lett. 1982 Sep 6;146(1):111–114. doi: 10.1016/0014-5793(82)80715-1. [DOI] [PubMed] [Google Scholar]
  2. Bernström K., Hammarström S. Metabolism of leukotriene D by porcine kidney. J Biol Chem. 1981 Sep 25;256(18):9579–9582. [PubMed] [Google Scholar]
  3. Borgeat P., Samuelsson B. Transformation of arachidonic acid by rabbit polymorphonuclear leukocytes. Formation of a novel dihydroxyeicosatetraenoic acid. J Biol Chem. 1979 Apr 25;254(8):2643–2646. [PubMed] [Google Scholar]
  4. Bray M. A., Cunningham F. M., Ford-Hutchinson A. W., Smith M. J. Leukotriene B4: a mediator of vascular permeability. Br J Pharmacol. 1981 Mar;72(3):483–486. doi: 10.1111/j.1476-5381.1981.tb11000.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bretz U., Baggiolini M. Biochemical and morphological characterization of azurophil and specific granules of human neutrophilic polymorphonuclear leukocytes. J Cell Biol. 1974 Oct;63(1):251–269. doi: 10.1083/jcb.63.1.251. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bryant R. W., Bailey J. M., Schewe T., Rapoport S. M. Positional specificity of a reticulocyte lipoxygenase. Conversion of arachidonic acid to 15-S-hydroperoxy-eicosatetraenoic acid. J Biol Chem. 1982 Jun 10;257(11):6050–6055. [PubMed] [Google Scholar]
  7. Camp R. D., Woollard P. M., Mallet A. I., Fincham N. J., Ford-Hutchinson A. W., Bray M. A. Neutrophil aggregating and chemokinetic properties of a 5,12,20-trihydroxy-6,8,10,14-eicosatetraenoic acid isolated from human leukocytes. Prostaglandins. 1982 May;23(5):631–641. doi: 10.1016/s0090-6980(82)80003-8. [DOI] [PubMed] [Google Scholar]
  8. Carmagnol F., Sinet P. M., Rapin J., Jerome H. Glutathione-S-transferase of human red blood cells; assay, values in normal subjects and in two pathological circumstances: hyperbilirubinemia and impaired renal function. Clin Chim Acta. 1981 Dec 9;117(2):209–217. doi: 10.1016/0009-8981(81)90040-1. [DOI] [PubMed] [Google Scholar]
  9. Chang W. C., Nakao J., Orimo H., Murota S. Effects of reduced glutathione on the 12-lipoxygenase pathways in rat platelets. Biochem J. 1982 Mar 15;202(3):771–776. doi: 10.1042/bj2020771. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Feinmark S. J., Lindgren J. A., Claesson H. E., Malmsten C., Samuelsson B. Stimulation of human leukocyte degranulation by leukotriene B4 and its omega-oxidized metabolites. FEBS Lett. 1981 Dec 21;136(1):141–144. doi: 10.1016/0014-5793(81)81233-1. [DOI] [PubMed] [Google Scholar]
  11. Frickhofen N., König W. Subcellular localization of the eosinophil chemotactic factor (ECF) and its inactivator in human polymorphonuclear leucocytes (PMN). Immunology. 1979 May;37(1):111–122. [PMC free article] [PubMed] [Google Scholar]
  12. Goetzl E. J. Selective feed-back inhibition of the 5-lipoxygenation of arachidonic acid in human T-lymphocytes. Biochem Biophys Res Commun. 1981 Jul 30;101(2):344–350. doi: 10.1016/0006-291x(81)91266-3. [DOI] [PubMed] [Google Scholar]
  13. Goldman D. W., Goetzl E. J. Specific binding of leukotriene B4 to receptors on human polymorphonuclear leukocytes. J Immunol. 1982 Oct;129(4):1600–1604. [PubMed] [Google Scholar]
  14. Gzarnetzki B. M., Konig W., Lichtenstein L. M. Release of eosinophil chemotactic factor from human polymorphonuclear neutrophils by calcium ionophore A23187 and phagocytosis. Nature. 1975 Dec 25;258(5537):725–726. doi: 10.1038/258725a0. [DOI] [PubMed] [Google Scholar]
  15. Habig W. H., Pabst M. J., Jakoby W. B. Glutathione S-transferases. The first enzymatic step in mercapturic acid formation. J Biol Chem. 1974 Nov 25;249(22):7130–7139. [PubMed] [Google Scholar]
  16. Henderson W. R., Jörg A., Klebanoff S. J. Eosinophil peroxidase-mediated inactivation of leukotrienes B4, C4, and D4. J Immunol. 1982 Jun;128(6):2609–2613. [PubMed] [Google Scholar]
  17. Ho P. P., Walters C. P., Sullivan H. R. A particulate arachidonate lipoxygenase in human blood platelets. Biochem Biophys Res Commun. 1976 May 23;76(2):398–405. doi: 10.1016/0006-291x(77)90738-0. [DOI] [PubMed] [Google Scholar]
  18. Jakoby W. B. The glutathione S-transferases: a group of multifunctional detoxification proteins. Adv Enzymol Relat Areas Mol Biol. 1978;46:383–414. doi: 10.1002/9780470122914.ch6. [DOI] [PubMed] [Google Scholar]
  19. Jakschik B. A., Harper T., Murphy R. C. Leukotriene C4 and D4 formation by particulate enzymes. J Biol Chem. 1982 May 25;257(10):5346–5349. [PubMed] [Google Scholar]
  20. Jim K., Hassid A., Sun F., Dunn M. J. Lipoxygenase activity in rat kidney glomeruli, glomerular epithelial cells, and cortical tubules. J Biol Chem. 1982 Sep 10;257(17):10294–10299. [PubMed] [Google Scholar]
  21. Jubiz W., Rådmark O., Lindgren J. A., Malmsten C., Samuelsson B. Novel leukotrienes: products formed by initial oxygenation of arachidonic acid at C-15. Biochem Biophys Res Commun. 1981 Apr 15;99(3):976–986. doi: 10.1016/0006-291x(81)91258-4. [DOI] [PubMed] [Google Scholar]
  22. Kroegel C., König W., Mollay C., Kreil G. Generation of the eosinophil chemotactic factor (ECF) from various cell types by melittin. Mol Immunol. 1981 Mar;18(3):227–236. doi: 10.1016/0161-5890(81)90089-4. [DOI] [PubMed] [Google Scholar]
  23. König W., Czarnetzki B. M., Lichtenstein L. M. Eosinophil chemotactic factor (ECF). II. Release from human polymorphonuclear leukocytes during phagocytosis. J Immunol. 1976 Jul;117(1):235–241. [PubMed] [Google Scholar]
  24. König W., Frickhofen N., Tesch H. Generation and secretion of eosinophilotactic activity from human polymorphonuclear neutrophils by various mechanisms of cell activation. Immunology. 1979 Apr;36(4):733–742. [PMC free article] [PubMed] [Google Scholar]
  25. König W., Tesch H., Frickhofen N. Generation and release of eosinophil chemotactic factor from human polymorphonuclear neutrophils by arachidonic acid. Eur J Immunol. 1978 Jun;8(6):434–437. doi: 10.1002/eji.1830080612. [DOI] [PubMed] [Google Scholar]
  26. Lilius E. M., Laakso S. A sensitive lipoxygenase assay based on chemiluminescence. Anal Biochem. 1982 Jan 1;119(1):135–141. doi: 10.1016/0003-2697(82)90676-5. [DOI] [PubMed] [Google Scholar]
  27. Maas R. L., Turk J., Oates J. A., Brash A. R. Formation of a novel dihydroxy acid from arachidonic acid by lipoxygenase-catalyzed double oxygenation in rat mononuclear cells and human leukocytes. J Biol Chem. 1982 Jun 25;257(12):7056–7067. [PubMed] [Google Scholar]
  28. Marcus A. J., Broekman M. J., Safier L. B., Ullman H. L., Islam N., Sherhan C. N., Rutherford L. E., Korchak H. M., Weissmann G. Formation of leukotrienes and other hydroxy acids during platelet-neutrophil interactions in vitro. Biochem Biophys Res Commun. 1982 Nov 16;109(1):130–137. doi: 10.1016/0006-291x(82)91575-3. [DOI] [PubMed] [Google Scholar]
  29. Morris H. R., Taylor G. W., Jones C. M., Piper P. J., Samhoun M. N., Tippins J. R. Slow reacting substances (leukotrienes): enzymes involved in their biosynthesis. Proc Natl Acad Sci U S A. 1982 Aug;79(16):4838–4842. doi: 10.1073/pnas.79.16.4838. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Narumiya S., Salmon J. A., Cottee F. H., Weatherley B. C., Flower R. J. Arachidonic acid 15-lipoxygenase from rabbit peritoneal polymorphonuclear leukocytes. Partial purification and properties. J Biol Chem. 1981 Sep 25;256(18):9583–9592. [PubMed] [Google Scholar]
  31. Nugteren D. H. Arachidonate lipoxygenase in blood platelets. Biochim Biophys Acta. 1975 Feb 20;380(2):299–307. doi: 10.1016/0005-2760(75)90016-8. [DOI] [PubMed] [Google Scholar]
  32. Ochi K., Yoshimoto T., Yamamoto S., Taniguchi K., Miyamoto T. Arachidonate 5-lipoxygenase of guinea pig peritoneal polymorphonuclear leukocytes. Activation by adenosine 5'-triphosphate. J Biol Chem. 1983 May 10;258(9):5754–5758. [PubMed] [Google Scholar]
  33. Orning L., Hammarström S. Kinetics of the conversion of leukotriene C by gamma-glutamyl transpeptidase. Biochem Biophys Res Commun. 1982 Jun 30;106(4):1304–1309. doi: 10.1016/0006-291x(82)91255-4. [DOI] [PubMed] [Google Scholar]
  34. Parker C. W., Aykent S. Calcium stimulation of the 5-lipoxygenase from RBL-1 cells. Biochem Biophys Res Commun. 1982 Dec 15;109(3):1011–1016. doi: 10.1016/0006-291x(82)92040-x. [DOI] [PubMed] [Google Scholar]
  35. Rand T. H., Turk J., Maas R. L., Colley D. G. Arachidonic acid metabolism of the murine eosinophil. II. Involvement of the lipoxygenase pathway in the response to the lymphokine eosinophil stimulation promoter. J Immunol. 1982 Sep;129(3):1239–1244. [PubMed] [Google Scholar]
  36. Rapoport S. M., Schewe T., Wiesner R., Halangk W., Ludwig P., Janicke-Höhne M., Tannert C., Hiebsch C., Klatt D. The lipoxygenase of reticulocytes. Purification, characterization and biological dynamics of the lipoxygenase; its identity with the respiratory inhibitors of the reticulocyte. Eur J Biochem. 1979 Jun 1;96(3):545–561. doi: 10.1111/j.1432-1033.1979.tb13068.x. [DOI] [PubMed] [Google Scholar]
  37. Rådmark O., Malmsten C., Samuelsson B., Clark D. A., Goto G., Marfat A., Corey E. J. Leukotriene A: stereochemistry and enzymatic conversion to leukotriene B. Biochem Biophys Res Commun. 1980 Feb 12;92(3):954–961. doi: 10.1016/0006-291x(80)90795-0. [DOI] [PubMed] [Google Scholar]
  38. Tate S. S., Meister A. gamma-Glutamyl transpeptidase: catalytic, structural and functional aspects. Mol Cell Biochem. 1981 Sep 25;39:357–368. doi: 10.1007/BF00232585. [DOI] [PubMed] [Google Scholar]
  39. Tesch H., König W. Phospholipase A2 and arachidonic acid: a common link in the generation of the eosinophil chemotactic factor (ECF) from human PMN by various stimuli. Scand J Immunol. 1980;11(4):409–418. doi: 10.1111/j.1365-3083.1980.tb00007.x. [DOI] [PubMed] [Google Scholar]
  40. Turk J., Maas R. L., Brash A. R., Roberts L. J., 2nd, Oates J. A. Arachidonic acid 15-lipoxygenase products from human eosinophils. J Biol Chem. 1982 Jun 25;257(12):7068–7076. [PubMed] [Google Scholar]
  41. Vanderhoek J. Y., Bryant R. W., Bailey J. M. Inhibition of leukotriene biosynthesis by the leukocyte product 15-hydroxy-5,8,11,13-eicosatetraenoic acid. J Biol Chem. 1980 Nov 10;255(21):10064–10066. [PubMed] [Google Scholar]
  42. Vanderhoek J. Y., Tare N. S., Bailey J. M., Goldstein A. L., Pluznik D. H. New role for 15-hydroxyeicosatetraenoic acid. Activator of leukotriene biosynthesis in PT-18 mast/basophil cells. J Biol Chem. 1982 Oct 25;257(20):12191–12195. [PubMed] [Google Scholar]
  43. Wallach D. P., Brown V. R. A novel preparation of human platelet lipoxygenase. Characteristics and inhibition by a variety of phenyl hydrazones and comparisons with other lipoxygenases. Biochim Biophys Acta. 1981 Feb 23;663(2):361–372. doi: 10.1016/0005-2760(81)90165-x. [DOI] [PubMed] [Google Scholar]

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