Biosynthesis of Hypusine in eIF-4D Precursors

  • M. H. Park
  • E. C. Wolff
  • A. Abbruzzese
  • J. E. Folk
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 250)


An unusual basic amino acid hypusine was discovered in 1971 in extracts of bovine brain by Shiba and coworkers, who determined its chemical structure as Nε -(4-amino-2-hydroxybutyl)lysine(1). Its distribution in various animal tissues as the free amino acid (2) and as a component of protein (3) was subsequently reported. Initial attempts to isolate any specific protein that contained hypusine were unsucessful, however (3). Ten years after the discovery of hypusine, we observed radiolabeling of a single cellular protein (Mr 183000, pI 5.3) upon incubation of human peripheral lymphocytes with [1, 8- 3H] spermidine or [2, 3- 3H] putrescine in the presence of the mitogen, phytohemagglutinin and identified the polyamine-derived radioactive component of this protein as hypusine (4).


Protein Substrate Ammonium Sulfate Fraction Bold Face Type Human Peripheral Lymphocyte Break Arrow 
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  1. 1.
    T. Shiba, H. Mizote, T. Kaneko, T. Nakajima, Y. Kakimoto, and I. Sano, Hypusine, a new amino acid occurring in bovine brain: Isolation and structural determination, Biochim. Biophys. Acta 244: 523 (1971).PubMedCrossRefGoogle Scholar
  2. 2.
    T. Nakajima, T. Matsubayashi, Y. Kakimoto, and I. Sano, Distribution of hypusine, N-(4-amino-2-hydroxybutyl)-2-6-diaminohexanoic acid, in mammalian organs, Biochim. Biophys. Acta 252: 92 (1971).PubMedCrossRefGoogle Scholar
  3. 3.
    N. Imaoka, and T. Nakajima, Hypusine, N6-(4-amino-2-hydroxybutyl)-2-6-diaminohexanoic acid, in tissue proteins of mammals, Biochim. Biophys. Acta, 320: 97 (1973).PubMedCrossRefGoogle Scholar
  4. 4.
    M.H. Park, H.L. Cooper, and J.E. Folk, Identification of hypusine, an unusual amino acid, in a protein from human lymphocytes and of spermidine as its biosynthetic precursor, Proc. Natl. Acad. Sci. USA, 78: 2869 (1981).PubMedCrossRefGoogle Scholar
  5. 5.
    H.L. Cooper, M.H. Park, J.E. Folk, B. Safer, and R. Braverman, Identification of the hypusine-containing protein Hy+ as translation initiation factor eIF-4D, Proc. Natl. Acad. Sci. USA, 80: 1854 (1983).PubMedCrossRefGoogle Scholar
  6. 6.
    M.H. Park, S.I. Chung, H.L. Cooper, and J.E. Folk, The mammalian hypusine containing protein, eukaryotic initiation factor 4D: Structural homology of this protein from several species, J. Biol. Chem. 259: 4562 (1984).Google Scholar
  7. 7.
    M.H. Park, T.Y. Liu, S.H. Neece, and W.J. Swiggard, Eukaryotic initiation factor 4D: Purification from human red blood cells and the sequence of the amino acids around its single hypusine residue, J. Biol. Chem. 261: 14515 (1986).PubMedGoogle Scholar
  8. 8.
    E.D. Gordon, R. Mora, S.C. Meredith, C. Lee and S.L. Lindquist, Eukaryotic initiation factor 4D, the hypusine containing protein is conserved among eukaryotes. J. Biol. Chem. 262: 16585 (1987).PubMedGoogle Scholar
  9. 9.
    M.H. Park Unpublished results.Google Scholar
  10. 10.
    K.Y. Chen and Y.C. Yang, Spermidine labels a 21, 000-dalton protein in Neurospora Crassa both in vivo and in vitro, The FASEB J. 2: A1563 (1988).Google Scholar
  11. 11.
    H.L. Cooper, M.H. Park, and J.E. Folk, Posttranslational formation of hypusine in a single major protein occurs generally in growing cells and is associated with activation of lymphocyte growth, Cell 29: 791 (1982).PubMedCrossRefGoogle Scholar
  12. 12.
    K.Y. Chen, An 18, 000 dalton protein metabolically labeled by polyamines in various mammalian cell lines, Biochim. Biophys. Acta 856: 395 (1983).CrossRefGoogle Scholar
  13. 13.
    B.M. Torrelio, M.A. Paz, and P.M. Gallop, Cellular proliferation and hypusine synthesis, Exp. Cell Res. 154: 454 (1983).CrossRefGoogle Scholar
  14. 14.
    E.W. Gerner, P.S. Mamont, A. Bernhardt and M. Siat, Post-translational modification of the protein-synthesis initiation factor eIF-4D by spermidine in rat hepatoma cells, Biochem. J. 239: 379 (1986).PubMedGoogle Scholar
  15. 15.
    A.A.M. Thomas, R. Benne, and H.O. Voorma, Initiation of eukaryotic protein synthesis, FEBS Letters 128: 177 (1981).PubMedCrossRefGoogle Scholar
  16. 16.
    M.H. Park, H.L. Cooper, and J.E. Folk, The biosynthesis of protein-bound hypusine [Nε.-(4-amino-2-hydroxybutyl)lysine]: Lysine as the amino acid precursor and the intermediate role of deoxyhypusine [N-(4-amino-butyl)lysine], J. Biol. Chem. 257: 7217 (1982).PubMedGoogle Scholar
  17. 17.
    M.H. Park, D.J. Liberato, A.L. Yergey, and J.E. Folk, The biosynthesis of hypusine [Nε-4-amino-2-hydroxybutyl)lysine]: Alignment of the butylamine segment and source of the secondary amino nitrogen, J. Biol. Chem. 259: 12123 (1984).PubMedGoogle Scholar
  18. 18.
    M.H. Park, and J.E., Folk, Biosynthetic labeling of hypusine in mammalian cells: carbon-hydrogen bond fissions revealed by dual labeling. J. Biol. Chem. 261: 14108 (1986).PubMedGoogle Scholar
  19. 19.
    A. Abbruzzese, M.H. Park, and J.E. Folk, Deoxyhypusine hydroxylase from rat testis: Partial purification and characterization, J. Biol. Chem. 261: 3085 (1986).PubMedGoogle Scholar
  20. 20.
    K.I. Kivirikko, and R. Myllyla, Post-translation enzymes in the biosynthesis of collagen: Intracellular enzymes, Methods Enzymol. 82: 245 (1982).PubMedCrossRefGoogle Scholar
  21. 21.
    M.H. Park, Regulation of biosynthesis of hypusine in Chinese hamster ovary cells: Evidence for eIF-4D precursor polypeptides. J. Biol. Chem. 262: 12730 (1987).PubMedGoogle Scholar
  22. 22.
    R.F. Duncan and J.W.B. Hershey, Changes in eIF-4D hypusine modification or abundance are not correlated with translational repression in HeLa cells, J. Biol. Chem. 261: 12903 (1986).PubMedGoogle Scholar
  23. 23.
    B.W. Metcalf, P. Bey, C. Danzin, M.J. Jung, P. Casara, and J.P. Vevert, Catalytic irreversible inhibition of mammalian ornithine decarboxylase (E.C. by substrate and product analogues, J. Am. Chem. Soc. 100: 2552 (1978).CrossRefGoogle Scholar
  24. 24.
    M.H. Park, Identification of an eukaryotic initiation factor 4D precursor in spermidine-depleted Chinese hamster ovary cells. J. Biol. Chem. 263: 7447 (1988).PubMedGoogle Scholar
  25. 25.
    J.I. Garrels, Two-dimensional gel electrophoresis and computer analysis of proteins synthesized by clonal cell lines, J. Biol. Chem. 254: 7961 (1979).PubMedGoogle Scholar
  26. 26.
    R.J. Murphey and E.W. Gerner, Hypusine formation in protein by a two-step process in cell lysates, J. Biol. Chem. 262: 15033 (1987).PubMedGoogle Scholar
  27. 27.
    M.H. Park and E.C. Wolff, Cell-free synthesis of deoxyhypusine: Separation of protein substrate and enzyme and Identification of 1, 3-diami-nopropane as a product of spermidine cleavage. J. Biol. Chem. (in press).Google Scholar

Copyright information

© Plenum Press, New York 1988

Authors and Affiliations

  • M. H. Park
    • 1
  • E. C. Wolff
    • 1
  • A. Abbruzzese
    • 2
  • J. E. Folk
    • 1
  1. 1.National Institute of Dental ResearchNIHBethesdaUSA
  2. 2.Department of Biochemistry and BiophysicsUniversity of NaplesItaly

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