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DOI: 10.1039/A901371G (Paper) Reference Section for: J. Chem. Soc., Perkin Trans. 1, 1999, 1015-1022

Total chemical synthesis of deglycosylated human erythropoietin

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Nicola Robertson and Robert Ramage

Abstract

For the first time the total chemical synthesis of deglycosylated human erythropoietin is described. Synthesis of this 166 amino acid protein, molecular weight 18 389 daltons, was achieved by automated synthesis on the solid phase by stepwise elongation with fluoren-9-ylmethoxycarbonyl amino acids. A novel coupling reagent ethyl 1-hydroxy-(1H)-1,2,3-triazole-4-carboxylate in conjunction with diisopropylcarbodiimide was employed for carboxy activation and tetrabenzo[a,c,g,i[hair space]]fluoren-17-ylmethoxycarbonyl, a hydrophobic, chromatographic probe which has an affinity for charcoal, aided the purification immensely.

References

  1. E. Goldwasser, Fed. Proc., 1975, 34, 2285 Search PubMed.
  2. S. E. Graber and S. B. Krantz, Annu. Rev. Med., 1978, 29, 51 CAS.
  3. A. J. Erslev, J. Wilson and J. Caro, J. Lab. Clin. Med., 1987, 109, 429 CAS.
  4. L. Jiang, A. Davison, G. Tennant and R. Ramage, Tetrahedron, 1998, 54, 14233 CrossRef CAS.
  5. N. Robertson, L. Jiang and R. Ramage, Tetrahedron, 1998, in press Search PubMed.
  6. I. Clark-Lewis, R. Aebersold, H. Ziltener, J. W. Schrader, L. E. Hood and S. B. H. Kent, Science, 1986, 231, 134 CAS.
  7. R. Ramage, J. Green, T. W. Muir, O. M. Ogunjobi, S. Love and K. Shaw, Biochem. J., 1994, 299, 151 CAS.
  8. R. Ramage and G. Raphy, Tetrahedron Lett., 1992, 33, 385 CrossRef CAS.
  9. A. R. Brown, S. L. Irving and R. Ramage, Tetrahedron Lett., 1993, 34, 7129 CrossRef CAS.
  10. K. Jacobs, C. Shoemaker, R. Rudersdorf, S. D. Neill, R. J. Kaufman, A. Mufson, J. Seehra, S. S. Jones, R. Hewick, E. F. Fritsch, M. Kawakita, T. Shmizu and T. Miyake, Nature, 1985, 313, 806 CAS.
  11. R. B. Merrifield, J. Am. Chem. Soc., 1963, 85, 2149 CrossRef CAS.
  12. S. S. Wang, J. Am. Chem. Soc., 1973, 95, 1328 CrossRef CAS.
  13. L. A. Carpino and G. Y. Han, J. Org. Chem., 1972, 37, 3404 CrossRef CAS.
  14. E. Atherton, C. J. Logan and R. C. Sheppard, J. Chem. Soc., Perkin Trans. 1., 1981, 538 RSC.
  15. K. M. Otteson, R. L. Nobel, P. D. Hoeprich, Jr., K. T. Shaw and R. Ramage, Applied Biosystems Res. News, June 1993 Search PubMed.
  16. H. Edeloch, Biochemistry, 1967, 6, 1948 CrossRef CAS.
  17. J.-P. Boissel, W. R. Lee, S. R. Presnell, F. E. Cohen and H. F. Bunn, J. Biol. Chem., 1993, 268, 15983 CAS.
  18. M. Le Maire, A. Viel and J. Moller, Anal. Biochem., 1989, 177, 50 CrossRef.
  19. H. Towbin, T. Staehelin and J. Gordon, Proc. Natl. Acad. Sci. USA, 1979, 76, 4350 CAS.
  20. U. K. Laemmli, Nature, 1970, 227, 680 CAS.
  21. J. D. Hayes, L. A. Kerr and A. D. Cronshaw, Biochem. J., 1989, 264, 437 CAS.
  22. C. H. Li, D. Yamashiro, D. Gospodarowicz, S. L. Kaplan and G. van Vliet, Proc. Natl. Acad. Sci. USA, 1983, 80, 2216 CAS.
  23. W. F. Heath and R. B. Merrifield, Proc. Natl. Acad. Sci. USA, 1986, 83, 6367 CAS.
  24. I. Clark-Lewis, B. Moser, A. Walz, M. Baggiolini, G. J. Scott and R. Abersold, Biochemistry, 1991, 30, 3128 CrossRef CAS.
  25. R. F. Nutt, S. F. Brady, P. L. Darke, T. M. Ciccarone, D. Colton, E. M. Nutt, J. A. Rodkey, C. D. Bennett, L. H. Waxman, I. S. Sigal, P. S. Anderson and A. F. Veber, Proc. Natl. Acad. Sci. USA, 1988, 85, 7129 CAS.