Biochemistry (Moscow)

, 74:75 | Cite as

High human GLUT1, GLUT2, and GLUT3 expression in Schizosaccharomyces pombe

  • Yuxin Yang
  • Zongli Hu
  • Zhizhao Liu
  • Yi Wang
  • Xuqing Chen
  • Guoping ChenEmail author


In this study, three subfamily members of the human 12-transmembrane-domain cell-surface receptors GLUT1, GLUT2, and GLUT3 were heterologously expressed in the fission yeast Schizosaccharomyces pombe utilizing GST-GLUT fusion proteins. These fusion proteins were driven by the full-length nmt1 promoter (Pnmt1) derived from S. pombe. The transcription levels of the GST-GLUT fusion proteins were very high upon induction by removing thiamine from the media. One-step purification of the recombinant fusion proteins was achieved by GST-affinity chromatography. Approximately 300 µg of highly purified fusion protein were obtained from 3 g of wet cell paste (1 liter of cell culture), indicating that human membrane proteins can be efficiently expressed and purified in the fission yeast. With its available extensive genetic information and ease of genetic manipulation, the fission yeast is potentially a highly efficient host to express eukaryotic membrane proteins.

Key words

GLUT Schizosaccharomyces pombe membrane protein expression GST-affinity chromatography 





Edinburgh minimal media


facilitated glucose transporters


glutathione S-transferase


polyvinylidene difluoride


yeast extract plus supplements medium


  1. 1.
    Miroux, B., and Walker, J. E. (1996) J. Mol. Biol., 260, 289–298.PubMedCrossRefGoogle Scholar
  2. 2.
    Sarramegna, V., Talmont, F., Demange, P., and Milon, A. (2003) Cell Mol. Life Sci., 60, 1529–1546.PubMedCrossRefGoogle Scholar
  3. 3.
    Dobrovetsky, E., Menendez, J., Edwards, A. M., and Koth, C. M. (2007) Methods, 41, 381–387.PubMedCrossRefGoogle Scholar
  4. 4.
    Niegowski, D., Hedren, M., Nordlund, P., and Eshaghi, S. (2006) Int. J. Biol. Macromol., 39, 83–87.PubMedCrossRefGoogle Scholar
  5. 5.
    Olson, A. L., and Pessin, J. E. (1996) Annu. Rev. Nutr., 16, 235–256.PubMedCrossRefGoogle Scholar
  6. 6.
    Mueckler, M. (1994) Eur. J. Biochem., 219, 713–725.PubMedCrossRefGoogle Scholar
  7. 7.
    Marger, M. D., and Saier, M. H. (1993) Trends Biochem. Sci., 18, 13–20.PubMedCrossRefGoogle Scholar
  8. 8.
    Brown, G. K. (2000) J. Inherit. Metab. Dis., 23, 237–246.PubMedCrossRefGoogle Scholar
  9. 9.
    Tohda, H., Okada, H., Giga-Hama, Y., Okayama, H., and Kumagai, H. (1994) Gene, 150, 275–280.PubMedCrossRefGoogle Scholar
  10. 10.
    Kaufer, N. F., Simanis, V., and Nurse, P. (1985) Nature, 318, 78–80.PubMedCrossRefGoogle Scholar
  11. 11.
    Giga-Hama, Y., Tohda, H., Okada, H., Owada, M. K., Okayama, H., and Kumagai, H. (1994) Biotechnology, 12, 400–404.PubMedCrossRefGoogle Scholar
  12. 12.
    Stoscheck, C. M. (1990) Meth. Enzymol., 182, 50–68.PubMedCrossRefGoogle Scholar
  13. 13.
    Maundrell, K. (1989) J. Biol. Chem., 265, 10857–10864.Google Scholar
  14. 14.
    Lu, Q., Bauer, J. C., and Greener, A. (1997) Gene, 200, 135–144.PubMedCrossRefGoogle Scholar
  15. 15.
    Weiss, H. M., and Grisshammer, R. (2002) Eur. J. Biochem., 269, 82–92.PubMedCrossRefGoogle Scholar
  16. 16.
    Tschantz, W. R., Pfeifer, N. D., Meade, C. L., Wang, L., Lanzetti, A., Kamath, A. V., Berlioz-Seux, F., and Hashim, M. F. (2008) Prot. Exp. Purif., 57, 163–171.CrossRefGoogle Scholar
  17. 17.
    Jidenko, M., Lenoir, G., Fuentes, J. M., le Maire, M., and Jaxel, C. (2006) Prot. Exp. Purif., 48, 32–42.CrossRefGoogle Scholar
  18. 18.
    Tate, C. G. (2001) FEBS Lett., 504, 94–98.PubMedCrossRefGoogle Scholar
  19. 19.
    Kunji, E. R., Chan, K. W., Slotboom, D. J., Floyd, S., O’Connor, R., and Monne, M. (2005) Curr. Opin. Biotechnol., 16, 546–551.PubMedCrossRefGoogle Scholar
  20. 20.
    Kjaerulff, S., and Jensen, M. R. (2005) Biochem. Biophys. Res. Commun., 336, 974–982.PubMedCrossRefGoogle Scholar
  21. 21.
    Russell, P., and Nurse, P. (1986) Cell, 45, 781–782.PubMedCrossRefGoogle Scholar
  22. 22.
    Brys, R., Nelles, L., van der Schueren, E., Silvestre, N., Huylebroeck, D., and Remacle, J. E. (1998) DNA Cell. Biol., 17, 349–358.PubMedCrossRefGoogle Scholar
  23. 23.
    Remacle, J. E., Albrecht, G., Brys, R., Braus, G. H., and Huylebroeck, D. (1997) EMBO J., 16, 5722–5729.PubMedCrossRefGoogle Scholar
  24. 24.
    Moreno, S., Sanchez, Y., and Rodriguez, L. (1990) Biochem. J., 267, 697–702.PubMedGoogle Scholar
  25. 25.
    Chappell, T. G., and Warren, G. J. (1989) Cell Biol., 109, 2693–2702.CrossRefGoogle Scholar
  26. 26.
    Parodi, A. J. (1999) Biochim. Biophys. Acta, 1426, 287–295.PubMedGoogle Scholar
  27. 27.
    Bureik, M., Schiffler, B., Hiraoka, Y., Vogel, F., and Bernhardt, R. (2002) Biochemistry, 41, 2311–2321.PubMedCrossRefGoogle Scholar
  28. 28.
    Tabuchi, M., Yoshida, T., Takegawa, K., and Kishi, F. (1999) Biochem. J., 344, 211–219.PubMedCrossRefGoogle Scholar
  29. 29.
    Tiede, A., Schubert, J., Nischan, C., Jensen, I., Westfall, B., Taron, C. H., Orlean, P., and Schmidt, R. E. (1998) Biochem. J., 334, 609–616.PubMedGoogle Scholar
  30. 30.
    Bayele, H. K., Eisenthal, R. S., and Towner, P. (2000) J. Biol. Chem., 275, 14217–14222.PubMedCrossRefGoogle Scholar
  31. 31.
    Moreno, S., Klar, A., and Nurse, P. (1991) Meth. Enzymol., 194, 795–823.PubMedCrossRefGoogle Scholar
  32. 32.
    Reinhard, G. (2006) Curr. Opin. Biotechnol., 17, 337–340.CrossRefGoogle Scholar
  33. 33.
    Schuck, S., Honsho, M., Ekroos, K., Shevchenko, A., and Simons, K. (2003) Proc. Natl. Acad. Sci. USA, 100, 5795–5800.PubMedCrossRefGoogle Scholar
  34. 34.
    Koronakis, V., Sharff, A., Koronakis, E., Luisi, B., and Hughes, C. (2000) Nature, 40, 5914–5919.Google Scholar
  35. 35.
    Lemieux, M. J., Song, J., Kim, M. J., Huang, Y., Villa, A., Auer, M., Li, X. D., and Wang, D. N. (2003) Protein Sci., 12, 2748–2756.PubMedCrossRefGoogle Scholar

Copyright information

© Pleiades Publishing, Ltd. 2009

Authors and Affiliations

  • Yuxin Yang
    • 1
  • Zongli Hu
    • 1
  • Zhizhao Liu
    • 1
  • Yi Wang
    • 1
  • Xuqing Chen
    • 1
    • 2
  • Guoping Chen
    • 1
    Email author
  1. 1.College of BioengineeringChongqing UniversityShapingbaChina
  2. 2.Beijing Research Center of Agro-BiotechnologyBeijingChina

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