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The structure of mammalian 15-lipoxygenase reveals similarity to the lipases and the determinants of substrate specificity

A Correction to this article was published on 01 March 1998

Abstract

Here we report the first structure of a mammalian 15-lipoxygenase. The protein is composed of two domains; a catalytic domain and a previously unrecognized β-barrel domain. The N-terminal β-barrel domain has topological and sequence identity to a domain in the mammalian lipases, suggesting that these domains may have similar functions in vivo. Within the C-terminal domain, the lipoxygenase substrate binding site is a hydrophobic pocket defined by a bound inhibitor. Arachidonic acid can be docked into this deep hydrophobic pocket with the methyl end extending down into the bottom of the pocket and the acid end tethered by a conserved basic residue on the surface of the enzyme. This structure provides a unifying hypothesis for the positional specificity of mammalian lipoxygenases.

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References

  1. Samuelsson, B., Dahlén, S.-E., Lingren, J.Å., Rouzer, C.A. & Serhan, C.N. Leukotrienes and Lipoxins: Structures, Biosynthesis, and Biological Effects. Science 237, 1171–1176 (1987).

    Article  CAS  Google Scholar 

  2. Yokomizo, T., Izumi, T., Chang, K., Takuwa, Y. & Shimizu, T. A G-protein coupled receptor for leukotriene B4 that mediates chemotaxis. Nature 387, 620–624 (1997).

    Article  CAS  Google Scholar 

  3. Devchand, P.R. et al. The PPARa-Leukotriene B4 pathway to inflammation control. Nature 384, 39–43 (1996).

    Article  CAS  Google Scholar 

  4. Forman, B.M., Chen, J. & Evans, R.M. Hypolipidemic drugs, polyunsaturated fatty acids, and eicosanoids are ligands for peroxisome proliferator-activated receptors alpha and delta. Proc. Nat. Acad. Sci. USA 94, 4312–4317 (1997).

    Article  CAS  Google Scholar 

  5. Israel, E., Cohn, J., Dubé, L. & Drazen, J.M. Effect of treatment with Zileuton, a 5-lipoxygenase inhibitor, in patients with asthma. J. Am. Med. Assn. 275, 931–936 (1996).

    Article  CAS  Google Scholar 

  6. Shannon, V.R., Stenson, W.F. & Holtzman, M.J. Induction of epithelial arachidonate 12-lipoxygenase at active sites of inflammatory bowel disease. Am. J. Physiol. 264, G104–111 (1993).

    CAS  PubMed  Google Scholar 

  7. Steinberg, D., Parthasarathy, S., Carew, T.E., Khoo, J.C. & Witztum, M.D. Beyond cholesterol: Modifications of low-density lipoprotein that increase its atherogenicity. N. Engl. J. Med. 320, 915–924 (1989).

    Article  CAS  Google Scholar 

  8. Yla-Herttuala, S. et al. Gene expression in macrophage-rich human atherosclerotic lesions. 15-lipoxygenase and acetyl low density lipoprotein receptor messenger RNA colocalize with oxidation specific lipid-protein adducts. J. Clin. Invest. 87, 1146–1152 (1991).

    Article  CAS  Google Scholar 

  9. Kühn, J., Belkner, J., Zaiss, S., Fahrenklemper, T. & Wohlfeil, S. Involvement of 15-lipoxygenase in early stages of atherosclerosis. J. Exp. Med. 179, 1903–1911 (1994).

    Article  Google Scholar 

  10. Ford-Hutchinson, A.W. Arachidonate 15-lipoxygenase; characteristics and potential biological significance. Eicosanoids 4, 65–74 (1991).

    CAS  PubMed  Google Scholar 

  11. The CCP4 Suite:Programs for protein crystallography. Acta Crystallogr. D50, 760–763 (1994).

  12. Minor, W. et al. Crystal structure of soybean lipoxygenase-1 at 1.4Å resolution. Biochemistry 35, 10687–10701 (1996).

    Article  CAS  Google Scholar 

  13. Boyington, J.C., Gaffney, B.J. & Amzel, L.M. The three-dimensional structure of an arachidonic acid 15-lipoxygenase. Science 260, 1482–1486 (1993).

    Article  CAS  Google Scholar 

  14. Funk, C.D. The molecular biology of mammalian lipoxygenases and the quest for eicosanoid functions using lipoxygenase-deficient mice. Biochem. Biophys. Acta 1304, 65–84 (1996).

    Article  Google Scholar 

  15. Prigge, S.T., Boyington, J.C., Gaffney, B.J. & Amzel, L.M. Structure conservation in lipoyxgenases: structural analysis of soybean lipoxygenase-1 and modeling of human lipoxygenases. Proteins 24, 275–291 (1996).

    Article  CAS  Google Scholar 

  16. Sigal, E. et al. Molecular cloning and primary structure of human 15-lipoxygenase. Biochem. Biophys. Res. Comm. 157, 457–464 (1988).

    Article  CAS  Google Scholar 

  17. Winkler, F.W., D′Arcy, A. & Hunziker, W. Structure of human pancreatic lipase. Nature 343, 771–774 (1990).

    Article  CAS  Google Scholar 

  18. Williams, S.E. et al. The carboxyl-terminal domain of lipoprotein lipase binds to the low density lipoprotein receptor related protein/a2-macroglobulin receptor (LRP) and mediates binding of normal very low density lipoproteins to LRP. J. Biol. Chem. 269, 8653–8658 (1994).

    CAS  PubMed  Google Scholar 

  19. Van Tilbeurgh, H., Sarda, L., Verger, R. & Cambillau, C. Structure of the pancreatic lipase-procolipase complex. Nature 359, 159–163 (1992).

    Article  CAS  Google Scholar 

  20. Carriere, F. et al. Pancreatic lipase structure-funtion relationships by domain exchange. Biochemistry 36, 239–248 (1997).

    Article  CAS  Google Scholar 

  21. Miller, D.K. et al. Identification and isolation of a membrane protein necessary for leukotriene production. Nature 343, 278–281 (1990).

    Article  CAS  Google Scholar 

  22. Dixon, R.A.F. et al. Requirement of a 5-lipoxygenase activating protein for leukotriene synthesis. Nature 343, 282–284 (1990).

    Article  CAS  Google Scholar 

  23. Zhang, Y., Gan, Q.-F., Pavel, E.G., Sigal, E. & Solomon, E.I. EPR definition of the non-heme ferric active sites of mammalian 15-lipoxygenase: major spectral difference relative to human 5-lipoxygenase and plant lipoxygenase and their ligand field origin. J. Am. Chem. Soc. 117, 7422–7427 (1995).

    Article  CAS  Google Scholar 

  24. Gorins, G., Kuhnert, L., Johnson, C.R. & Marnett, L.J. (Carboxyalkyl)benzyl propargyl ethers as selective inhibitors of leukocyte-type 12-lipoxygenases. J. Med. Chem. 39, 4871–4878 (1996).

    Article  CAS  Google Scholar 

  25. Corey, E.J. & Nagata, R. Evidence in favor of an organoiron-mediated pathway for lipoxygenation of fatty acids by soybean lipoxygenase. J. Am. Chem. Soc. 109, 8107–8108 (1987).

    Article  CAS  Google Scholar 

  26. de Groot, J.J. et al. Demonstration by EPR spectroscopy of the functional role of iron in soybean lipoxygenase-1. Biochim. Biophys. Acta 377, 71–79 (1975).

    Article  CAS  Google Scholar 

  27. Glickman, M.H. & Klinman, J.P. Lipoxygenase reaction mechanism: demonstration that hydrogen abstraction from substrate predeces dioxygen binding during catalytic turnover. Biochemistry 35, 12881–12892 (1996).

    Article  Google Scholar 

  28. Gan, Q.F., Browner, M.F., Sloane, D.L. & Sigal, E. Defining the arachidonic acid binding site of human 15-lipoxygenase. Molecular modeling and mutagenesis. J. Biol. Chem. 271, 25412–25418 (1996).

    Article  CAS  Google Scholar 

  29. Sloane, D.L., Leung, R., Craik, C.S. & Sigal, E. A primary determinant for lipoxygenase positional specificity. Nature 354, 149–152 (1991).

    Article  CAS  Google Scholar 

  30. Chen, X.S. & Funk, C.D. Structure-function properties of human platelet 12-lipoxygenase: chimeric enzyme and in vitro mutagenesis studies. Faseb J. 7, 694–701 (1993).

    Article  CAS  Google Scholar 

  31. Borngräber, S., Kuban, R.-J., Anton, M. & Kühn, H. Phenylalanine 353 is a primary determinant for the positional specificity of Mammalian 15-lipoxygenases. J. Mol. Biol. 264, 1145–1153 (1996).

    Article  Google Scholar 

  32. Brash, A.R., Boeglin, W.E. & Chang, M.S. Discovery of a second 15S-lipoxygenase in humans. Proc. Natl. Acad. Sci. USA 94, 6148–6152 (1997).

    Article  CAS  Google Scholar 

  33. Schewe, T., Rapoport, S.M. & Kuhn, H. Enzymology and Physiology of reticulocyte lipoxygenase: comparison with other lipoxygenases. in Advances in enzymology Vol. 58 (ed. Meister, A.) 191–272 (John Wiley and Sons, New York, 1986).

    Google Scholar 

  34. Picot, D., Loll, P.J. & Garavito, R.M. The X-ray crystal structure of the membrane protein prostaglandin H2 synthase-1. Nature 367, 243–249 (1994).

    Article  CAS  Google Scholar 

  35. Luong, C. et al. Flexibility of the NSAID binding site in the structure of human cyclooxygenase-2. Nature Struct. Biol. 3, 927–933 (1996).

    Article  CAS  Google Scholar 

  36. Li, H. & Poulos, T.L. The structure of the cytochrome p450BM-3 haem domain complexed with the fatty acid substrate palmitoleic acid. Nature Struct. Biol. 4, 140–146 (1997).

    Article  CAS  Google Scholar 

  37. Sloane, D.L. et al. Purification and crystallization of 15-lipoxygenase from rabbit reticulocytes. Biochem. Biophys. Res. Comm. 173, 507–513 (1990).

    Article  CAS  Google Scholar 

  38. Otwinowski, Z. Data collection and processing. in Proceedings of the CCP4 Study Weekend: Data Collection and Processing (eds Sawyer, L., Isaacs, N. & Baily, S.) 56–62 (Science and Engineering Research Council, Daresbury Laboratory, Daresbury, England, 1993).

    Google Scholar 

  39. Furey, W. & Swamintha, S. PHASES. in Macromolecular crystallography Vol. 276 (eds Carter, C. & Sweet, R.) (Academic Press, Orlando, 1996).

    Google Scholar 

  40. Muller, K. MOLOC: A molecular modeling program. Bull. Soc. Chim. 97, 655–667 (1988).

    Article  Google Scholar 

  41. Brünger, A.T. X-PLOR: version 3.1 (Yale Press, New Haven, 1987).

    Google Scholar 

  42. Holm, L. & Sander, C. Dali ver. 2.0. J. Mol. Biol. 233, 123–138 (1993).

    Article  CAS  Google Scholar 

  43. Nicholls, A., Sharp, K. & Honig, B. Protein folding and association: insights from the interfacical and thermodynamic properties of hydrocarbons. Proteins 11, 281–296 (1991).

    Article  CAS  Google Scholar 

Download references

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Gillmor, S., Villaseñor, A., Fletterick, R. et al. The structure of mammalian 15-lipoxygenase reveals similarity to the lipases and the determinants of substrate specificity. Nat Struct Mol Biol 4, 1003–1009 (1997). https://doi.org/10.1038/nsb1297-1003

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