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. 1990 Oct;87(19):7477–7481. doi: 10.1073/pnas.87.19.7477

Cloning of the cDNA for human 12-lipoxygenase.

T Izumi 1, S Hoshiko 1, O Rådmark 1, B Samuelsson 1
PMCID: PMC54770  PMID: 2217179

Abstract

A full-length cDNA clone encoding 12-lipoxygenase (arachidonate:oxygen 12-oxidoreductase, EC 1.13.11.31) was isolated from a human platelet cDNA library by using a cDNA for human reticulocyte 15-lipoxygenase as probe for the initial screening. The cDNA had an open reading frame encoding 662 amino acid residues with a calculated molecular weight of 75,590. Three independent clones revealed minor heterogeneities in their DNA sequences. Thus, in three positions of the deduced amino acid sequence, there is a choice between two different amino acids. The deduced sequence from the clone plT3 showed 65% identity with human reticulocyte 15-lipoxygenase and 42% identity with human leukocyte 5-lipoxygenase. The 12-lipoxygenase cDNA recognized a 3.0-kilobase mRNA species in platelets and human erythroleukemia cells (HEL cells). Phorbol 12-tetradecanoyl 13-acetate induced megakaryocytic differentiation of HEL cells and 12-lipoxygenase activity and increased mRNA for 12-lipoxygenase. The identity of the cloned 12-lipoxygenase was assured by expression in a mammalian cell line (COS cells). Human platelet 12-lipoxygenase has been difficult to purify to homogeneity. The cloning of this cDNA will increase the possibilities to elucidate the structure and function of this enzyme.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Balcarek J. M., Theisen T. W., Cook M. N., Varrichio A., Hwang S. M., Strohsacker M. W., Crooke S. T. Isolation and characterization of a cDNA clone encoding rat 5-lipoxygenase. J Biol Chem. 1988 Sep 25;263(27):13937–13941. [PubMed] [Google Scholar]
  2. Brash A. R. A review of possible roles of the platelet 12-lipoxygenase. Circulation. 1985 Oct;72(4):702–707. doi: 10.1161/01.cir.72.4.702. [DOI] [PubMed] [Google Scholar]
  3. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  4. Claeys M., Kivits G. A., Christ-Hazelhof E., Nugteren D. H. Metabolic profile of linoleic acid in porcine leukocytes through the lipoxygenase pathway. Biochim Biophys Acta. 1985 Oct 23;837(1):35–51. doi: 10.1016/0005-2760(85)90083-9. [DOI] [PubMed] [Google Scholar]
  5. Dixon R. A., Jones R. E., Diehl R. E., Bennett C. D., Kargman S., Rouzer C. A. Cloning of the cDNA for human 5-lipoxygenase. Proc Natl Acad Sci U S A. 1988 Jan;85(2):416–420. doi: 10.1073/pnas.85.2.416. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fleming J., Thiele B. J., Chester J., O'Prey J., Janetzki S., Aitken A., Anton I. A., Rapoport S. M., Harrison P. R. The complete sequence of the rabbit erythroid cell-specific 15-lipoxygenase mRNA: comparison of the predicted amino acid sequence of the erythrocyte lipoxygenase with other lipoxygenases. Gene. 1989 Jun 30;79(1):181–188. doi: 10.1016/0378-1119(89)90103-0. [DOI] [PubMed] [Google Scholar]
  7. Funk C. D., Gunne H., Steiner H., Izumi T., Samuelsson B. Native and mutant 5-lipoxygenase expression in a baculovirus/insect cell system. Proc Natl Acad Sci U S A. 1989 Apr;86(8):2592–2596. doi: 10.1073/pnas.86.8.2592. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Grossi I. M., Fitzgerald L. A., Umbarger L. A., Nelson K. K., Diglio C. A., Taylor J. D., Honn K. V. Bidirectional control of membrane expression and/or activation of the tumor cell IRGpIIb/IIIa receptor and tumor cell adhesion by lipoxygenase products of arachidonic acid and linoleic acid. Cancer Res. 1989 Feb 15;49(4):1029–1037. [PubMed] [Google Scholar]
  9. Hamberg M., Samuelsson B. Prostaglandin endoperoxides. Novel transformations of arachidonic acid in human platelets. Proc Natl Acad Sci U S A. 1974 Sep;71(9):3400–3404. doi: 10.1073/pnas.71.9.3400. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hansbrough J. R., Takahashi Y., Ueda N., Yamamoto S., Holtzman M. J. Identification of a novel arachidonate 12-lipoxygenase in bovine tracheal epithelial cells distinct from leukocyte and platelet forms of the enzyme. J Biol Chem. 1990 Jan 25;265(3):1771–1776. [PubMed] [Google Scholar]
  11. Kozak M. Compilation and analysis of sequences upstream from the translational start site in eukaryotic mRNAs. Nucleic Acids Res. 1984 Jan 25;12(2):857–872. doi: 10.1093/nar/12.2.857. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Long M. W., Heffner C. H., Williams J. L., Peters C., Prochownik E. V. Regulation of megakaryocyte phenotype in human erythroleukemia cells. J Clin Invest. 1990 Apr;85(4):1072–1084. doi: 10.1172/JCI114538. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Matsumoto T., Funk C. D., Rådmark O., Hög J. O., Jörnvall H., Samuelsson B. Molecular cloning and amino acid sequence of human 5-lipoxygenase. Proc Natl Acad Sci U S A. 1988 Jan;85(1):26–30. doi: 10.1073/pnas.85.1.26. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Nakao J., Ooyama T., Ito H., Chang W. C., Murota S. Comparative effect of lipoxygenase products of arachidonic acid on rat aortic smooth muscle cell migration. Atherosclerosis. 1982 Sep;44(3):339–342. doi: 10.1016/0021-9150(82)90008-9. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. Piomelli D., Volterra A., Dale N., Siegelbaum S. A., Kandel E. R., Schwartz J. H., Belardetti F. Lipoxygenase metabolites of arachidonic acid as second messengers for presynaptic inhibition of Aplysia sensory cells. Nature. 1987 Jul 2;328(6125):38–43. doi: 10.1038/328038a0. [DOI] [PubMed] [Google Scholar]
  17. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
  18. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Seed B. An LFA-3 cDNA encodes a phospholipid-linked membrane protein homologous to its receptor CD2. 1987 Oct 29-Nov 4Nature. 329(6142):840–842. doi: 10.1038/329840a0. [DOI] [PubMed] [Google Scholar]
  20. Shibata D., Steczko J., Dixon J. E., Andrews P. C., Hermodson M., Axelrod B. Primary structure of soybean lipoxygenase L-2. J Biol Chem. 1988 May 15;263(14):6816–6821. [PubMed] [Google Scholar]
  21. Sigal E., Craik C. S., Highland E., Grunberger D., Costello L. L., Dixon R. A., Nadel J. A. Molecular cloning and primary structure of human 15-lipoxygenase. Biochem Biophys Res Commun. 1988 Dec 15;157(2):457–464. doi: 10.1016/s0006-291x(88)80271-7. [DOI] [PubMed] [Google Scholar]
  22. Tabilio A., Rosa J. P., Testa U., Kieffer N., Nurden A. T., Del Canizo M. C., Breton-Gorius J., Vainchenker W. Expression of platelet membrane glycoproteins and alpha-granule proteins by a human erythroleukemia cell line (HEL). EMBO J. 1984 Feb;3(2):453–459. doi: 10.1002/j.1460-2075.1984.tb01827.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Takahashi Y., Ueda N., Yamamoto S. Two immunologically and catalytically distinct arachidonate 12-lipoxygenases of bovine platelets and leukocytes. Arch Biochem Biophys. 1988 Nov 1;266(2):613–621. doi: 10.1016/0003-9861(88)90294-9. [DOI] [PubMed] [Google Scholar]
  24. Turk J., Rand T. H., Maas R. L., Lawson J. A., Brash A. R., Roberts L. J., 2nd, Colley D. G., Oates J. A. Identification of lipoxygenase products from arachidonic acid metabolism in stimulated murine eosinophils. Biochim Biophys Acta. 1983 Jan 7;750(1):78–90. doi: 10.1016/0005-2760(83)90206-0. [DOI] [PubMed] [Google Scholar]
  25. Ueda N., Hiroshima A., Natsui K., Shinjo F., Yoshimoto T., Yamamoto S., Ii K., Gerozissis K., Dray F. Localization of arachidonate 12-lipoxygenase in parenchymal cells of porcine anterior pituitary. J Biol Chem. 1990 Feb 5;265(4):2311–2316. [PubMed] [Google Scholar]
  26. Yoshimoto T., Miyamoto Y., Ochi K., Yamamoto S. Arachidonate 12-lipoxygenase of porcine leukocyte with activity for 5-hydroxyeicosatetraenoic acid. Biochim Biophys Acta. 1982 Dec 13;713(3):638–646. [PubMed] [Google Scholar]
  27. Yoshimoto T., Suzuki H., Yamamoto S., Takai T., Yokoyama C., Tanabe T. Cloning and sequence analysis of the cDNA for arachidonate 12-lipoxygenase of porcine leukocytes. Proc Natl Acad Sci U S A. 1990 Mar;87(6):2142–2146. doi: 10.1073/pnas.87.6.2142. [DOI] [PMC free article] [PubMed] [Google Scholar]

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