Biosynthesis of Leukotriene B4 in Human Leukocytes

Demonstration of a Calcium-Dependent 5-Lipoxygenase
  • Rodger M. McMillan
  • David J. Masters
  • Wayne W. Sterling
  • Peter R. Bernstein
Part of the GWUMC Department of Biochemistry Annual Spring Symposia book series (GWUN)


Arachidonate metabolism via 5-lipoxygenase leads to formation of a family of biologically active molecules, the leukotrienes. The first step in leukotriene biosynthesis is catalyzed by the enzyme 5-lipoxygenase and results in formation of 5-hydroperoxy-6, 8, 11, 14-eicosatetraenoic acid (5-HPETE), which may be converted enzymatically (via the action of a peroxidase) or nonenzymically to the corresponding hydroxy acid, 5-hydroxy-6, 8, 11, 14-eicosatetraenoic acid (5-HETE). Alternatively, 5-HPETE may undergo enzymatic dehydration to form an unstable epoxide, 5, 6-epoxy-7, 9, 11, 14-eicosatetraenoic acid, which is termed leukotriene A4 (LTA4). Enzymatic hydrolysis of LTA4 yields 5S, 12R-dihydroxy-6, 14-cis-8, 10-trans-eicosatetraenoic acid (leukotriene B4, LTB4), which is a potent, stereospecific activator of a variety of human neutrophil responses, including chemokinesis, chemotaxis, and aggregation (Ford-Hutchinson et al., 1980; Goetzl and Pickett, 1981; Malmsten et al., 1980).


Human Leukocyte Epoxide Hydrolase Microsomal Epoxide Hydrolase Arachidonate Metabolism Synthetic LTB4 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Borgeat, P., and Samuelsson, B., 1979a, Arachidonic acid metabolism in polymorphonuclear leukocytes: Effects of ionophore A23187, Proc. Natl. Acad. Sci. U.S.A. 76:2148–2152.PubMedCrossRefGoogle Scholar
  2. Borgeat, P., and Samuelsson, B., 1979b, Metabolism of arachidonic acid in polymorphonuclear leukocytes. Structural analysis of novel hydroxylated compounds, J. Biol. Chem. 254:7865–7869.PubMedGoogle Scholar
  3. Borgeat, P., de LaClos, B.F., Picard, S., Drapeau, J., Vallerand, P., and Corey, E.J., 1982, Studies on the mechanism of formation of the 5S, 12S-dihydroxy-6, 8, 10, 14(E, Z, E, Z)-icosatetraenoic acid in leukocytes, Prostaglandins 23:713–724.PubMedCrossRefGoogle Scholar
  4. Clancy, R.M., Dahinden, C.A., and Hugli, T.E., 1983, 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. 80:7200–7204.PubMedCrossRefGoogle Scholar
  5. Corey, E.J., Marfat, A., Munroe, J., Kim, K.S., Hopkins, P.B., and Brion, F., 1981, A stereocontrolled and effective synthesis of leukotriene B, Tetrahedron Lett. 22:1077–1080.CrossRefGoogle Scholar
  6. Fitzpatrick, F., Haeggstrom, J., Granstrom, E., and Samuelsson, B., 1983, Metabolism of leukotriene A4 by an enzyme in blood plasma: A possible leukotactic mechanism, Proc. Natl. Acad. Sci. U.S.A. 80:5425–5429.PubMedCrossRefGoogle Scholar
  7. Ford-Hutchinson, A.W., Bray, M.A., Doig, M.V., Shipley, M.E., and Smith, M.J.H., 1980, Leukotriene B, a potent chemokinetic and aggregating substance released from polymorphonuclear leukocytes, Nature 286:264–265.PubMedCrossRefGoogle Scholar
  8. Ford-Hutchinson, A.W., Bray, M.A., Cunningham, F.M., Davidson, E.M., and Smith, M.J.H., 1981, Isomers of leukotriene B4 possess different biological properties, Prostaglandins 21:143–152.PubMedCrossRefGoogle Scholar
  9. Ford-Hutchinson, A.W., Rackham, A., Zamboni, R., Rokach, J., and Roy, S., 1983, Comparative biological properties of synthetic leukotriene B4 and its co-oxidation products, Prostaglandins 25:29–37.PubMedCrossRefGoogle Scholar
  10. Galliard, T., and Chan, H. W.-S., 1980, Lipoxygenases, in: The Biochemistry of Plants, Vol. 4, Academic Press, New York, pp. 131–161.Google Scholar
  11. Gill, S.S., and Hammock, B.D., 1980, Distribution and properties of a mammalian soluble epoxide hydrase, Biochem. Pharmacol. 29:389–395.PubMedCrossRefGoogle Scholar
  12. Goetzl, E.J., and Pickett, W.C., 1981, Novel structural determinants of the human neutrophil chemotactic activity of leukotriene B, J. Exp. Med. 153:482–487.PubMedCrossRefGoogle Scholar
  13. Hammock, B.D., and Hasagawa, L.S., 1983, Differential substrate selectivity of murine hapatic cytosolic and microsomal epoxide hydrolases, Biochem. Pharmacol. 32:1155–1164.PubMedCrossRefGoogle Scholar
  14. Hansson, G., Lindgren, J.A., Dahlen, S.-E., Hedquist, P., and Samuelsson, B., 1981, Identification and biological activity of novel ω-oxidized metabolites of leukotriene B4 from human leukocytes, FEBS Lett. 130:107–112.PubMedCrossRefGoogle Scholar
  15. Henson, P.M., 1971, The immunological release of constituents from neutrophil leukocytes. II. Mechanisms of release during phagocytosis, and adherence to non-phagocytosable surfaces, J. Immunol. 107:1547–1557.PubMedGoogle Scholar
  16. Jakschik, B.A., and Kuo, C.G., 1983, Subcellular localisation of leukotriene-forming enzymes, Adv. Prostaglandin Thromboxane Leukotriene Res. 11:141–145.Google Scholar
  17. Jakschik, B.A., Sun, F.F., Lee, L.-H., and Steinhoff, M.M., 1980a, Calcium stimulation of a novel lipoxygenase, Biochem. Biophys. Res. Commun. 95:103–110.PubMedCrossRefGoogle Scholar
  18. Jakschik, B.A., Sams, A.R., Sprecher, H., and Needleman, P., 1980b, Fatty acid structural requirements for leukotriene biosynthesis, Prostaglandins 20:401–410.PubMedCrossRefGoogle Scholar
  19. Koshihara, Y., Murota, S.-I., Petasis, N.A., and Nicolaou, K.C., 1982, Selective inhibition of 5-lipoxygenase by 5,6-methanoleukotriene A4, a stable analogue of leukotriene A4, FEBS Lett. 143:13–16.CrossRefGoogle Scholar
  20. Kreisle, R.A., and Parker, C.W., 1983, Specific binding of leukotriene B4 to a receptor on human polymorphonuclear leukocytes, J. Exp. Med. 157:628–641.PubMedCrossRefGoogle Scholar
  21. Lindgren, J.-A., Hansson, G., and Samuelsson, B., 1981, Formation of novel hydroxylated eicosatetraenoic acids in preparations of human polymorphonuclear leukocytes, FEBS Lett. 128:329–335.PubMedCrossRefGoogle Scholar
  22. Maduskie, T. P., and Willard, A. K., 1983, 17th Middle Atlantic Regional Meeting of the American Chemical Society. White Haven, PA, p. 162.Google Scholar
  23. Malmsten, C., Palmblad, J., Uden, A.-M., Radmark, O., Engstedt, L., and Samuelsson, B., 1980, Leukotriene B4: A highly potent and stereospecific factor stimulating migration of polymorphonuclear leukocytes, Acta Physiol. Scand. 110:449–451.PubMedCrossRefGoogle Scholar
  24. Maycock, A.L., Anderson, M.S., DeSousa, D.M., and Kuehl, F.A., 1982, Leukotriene A4: Preparation and enzymatic conversion in a cell-free system to leukotriene B4, J. Biol. Chem. 257:13911–13914.PubMedGoogle Scholar
  25. Murphy, R.C., Hammarström, S., and Samuelsson, B., 1979, Leukotriene C: A slow reacting substance (SRS) from mouse mastocytoma cells, Proc. Natl. Acad. Sci. U.S.A. 76:4275–4279.PubMedCrossRefGoogle Scholar
  26. Narumiya, S., Salmon, J. A., Flower, R. J., and Vane, J. R., 1982, Purification and properties of arachidonate-15-lipoxygenase from rabbit peritoneal polymorphonuclear leukocytes, in: Leukotrienes and Other Lipoxygenase Products (B. Samuelsson and R. Paoletti, eds.), Raven Press, New York, pp. 77-82.Google Scholar
  27. Nugteren, D.H., 1975, Arachidonate lipoxygenase in blood platelets, Biochim. Biophys. Acta 380:299–307.PubMedCrossRefGoogle Scholar
  28. Ochi, K., Yoshimoto, T., Yamamoto, S., Taniguchi, K., and Miyamoto, T., 1983, Arachidonate 5-lipoxygenase of guinea pig peritoneal polymorphonuclear leukocytes. Activation by adenosine 5′-triphosphate, J. Biol. Chem. 258:5574–5578.Google Scholar
  29. Oliw, E.H., Guengerich, F.P., and Oates, J.A., 1982, Oxygenation of arachidonic acid by hepatic monooxygenases. Isolation and metabolism of four epoxide intermediates, J. Biol. Chem. 257:3771–3781.PubMedGoogle Scholar
  30. Ota, K., and Hammock, B.D., 1980, Cytosolic and microsomal epoxide hydrolases: Differential properties in mammalian liver, Science 207:1479–1480.PubMedCrossRefGoogle Scholar
  31. Palmer, R.M.J., and Salmon, J.A., 1983, Release of leukotriene B4 from human neutrophils and its relationship to degranulation induced by N-formyl-methionyl-leucyl-phenylalanine, serum-treated zymosan and the ionophore A23187, Immunology 50:65–73.PubMedGoogle Scholar
  32. Parker, C.W., and Aykent, S., 1982, Calcium stimulation of the 5-lipoxygenase from RBL-1 cells, Biochem. Biophys. Res. Commun. 109:1011–1016.PubMedCrossRefGoogle Scholar
  33. Powell, W.S., 1984, Properties of leukotriene B4 20-hydroxylase from polymorphonuclear leukocytes, J. Biol. Chem. 259:3082–3089.PubMedGoogle Scholar
  34. Rae, S.A., Davidson, E.M., and Smith, M.J.H., 1982, Leukotriene B4, an inflammatory mediator in gout, Lancet 2:1122–1123.PubMedCrossRefGoogle Scholar
  35. Rokach, J., Zamboni, R., Lau, C.-K., and Gwriden, Y., 1981, The stereospecific synthesis of leukotriene A4 (LTA4), 5-epi-LTA4, 6-epi-LTA4 and 5-epi, 6-epi-LTA4, Tetrahedron Lett. 22:2759–2762.CrossRefGoogle Scholar
  36. Samuelsson, B., 1983, Leukotrienes: A new class of mediators of immediate hypersensitivity reactions and inflammation, Adv. Prostaglandin Thromboxane Leukotriene Res. 11:1–13.Google Scholar
  37. Smith, M.J.H., 1981, Leukotriene B4, Gen. Pharmacol. 12:211–216.PubMedCrossRefGoogle Scholar
  38. Yee, Y. K., 1984, A short and efficient synthesis of (2S, 3R)-oxido-5-cis-undecen-1-al. A total synthesis of LTB4. 14th Northeast Regional Meeting of the American Chemical Society. Google Scholar
  39. Yoshimoto, T., Miyamoto, Y., Ochi, K., and Yamamoto, S., 1982, Arachidonate 12-lipoxygenase of porcine leukocytes with activity for 5-hydroxyeicosatetraenoic acid, Biochim. Biophys. Acta 713:638–646.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1985

Authors and Affiliations

  • Rodger M. McMillan
    • 1
  • David J. Masters
    • 1
  • Wayne W. Sterling
    • 1
  • Peter R. Bernstein
    • 2
  1. 1.ICI Pharmaceuticals DivisionMeresideMacclesfield, CheshireUK
  2. 2.Medicinal Chemistry Department, Stuart PharmaceuticalsDivision of ICI Americas Inc.WilmingtonUSA

Personalised recommendations