Journal of the Korean Magnetic Resonance Society (한국자기공명학회논문지)

Korean Magnetic Resonance Society (한국자기공명학회)
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Expression and Purification of Unstructured Protein, IMUP-1, using Chaperone Co-expression System for NMR Study

  • Received : 2013.03.21
  • Accepted : 2013.06.10
  • Published : 2013.06.20
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Abstract

Immortalization-upregulated protein-1 (IMUP-1) genes have been cloned and are known to be involved in SV40-mediated immortalization. IMUP-1 gene is highly expressed in various cancer cell lines and tumors, suggesting the possibility that they might be involved in tumorigenicity. Previously, there were several problems for overexpression of IMUP-1 in bacterial expression systems including low solubility and aggregation due to unstructured property. To investigate the structural properties, it is necessary to obtain lots of pure and soluble proteins. Accordingly, the co-expression systems of bacterial chaperone proteins, GroEL-GroES, were used to increase solubility of IMUP-1. From the analysis of NMR and CD experiment data, it is suggested that the protein adopt typical the random coil properties in solution.

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References

  1. Hayflick, L., Moorhead, P. S. Cell Res. 25, 585. (1961). https://doi.org/10.1016/0014-4827(61)90192-6
  2. Hayflick, L. Cell Res. 37, 614. (1965). https://doi.org/10.1016/0014-4827(65)90211-9
  3. Jha, K. K., Banga, S., Palejwala, V., Ozer, H. L. Exp. Cell Res. 245, 1. (1998). https://doi.org/10.1006/excr.1998.4272
  4. Wright, W. E., Pereira-Smith, O. M., Shay, J. W. Mol. Ell. Biol. 9, 3088. (1989).
  5. Reddel, R. R. Ann. New York Acad. Sci. 854, 8. (1998). https://doi.org/10.1111/j.1749-6632.1998.tb09887.x
  6. Powell, A. J., Darmon, A. J., Gonos, E. S., Lan, E. W., Peden, K. W., Jat, P. S. Oncogene. 18, 7343. (1999). https://doi.org/10.1038/sj.onc.1203154
  7. J. K. Kim, R. Roland, I. Yoshiko, K. Seishi. Gene. 257, 327. (2000). https://doi.org/10.1016/S0378-1119(00)00414-5
  8. Z. Y. Ryoo, B. K. Jung, S. R. Lee, M. O. Kim, S. H. Kim, H. J. Kim, J. Y. Ahn, T. H. Lee, Y. H. Cho, J. H. Park, J. K. Kim. Biochem Biophys Res Commun. 349, 995. (2006). https://doi.org/10.1016/j.bbrc.2006.08.137
  9. J. K. Kim, H. J. An, N. K. Kim, J. Y. Ahn, K. S. Kim, Y. J. Kang, J. J. Ko, D. Oh, C. Lee, S. J. Kim, K. Y. Cha. Anticancer Res. 23, 4709. (2003).
  10. Cortazzo, P., Cervenansky, C., Marin, M., Reiss, C., Ehrlich, R., & Deana, A. Biochemical and Biophysical Research Communications. 293, 537. (2002). https://doi.org/10.1016/S0006-291X(02)00226-7
  11. Johnson, J. L., & Craig, E. A. Cell. 90,201. (1997).. https://doi.org/10.1016/S0092-8674(00)80327-X
  12. Horwich, A. L., Low, K. B., Fenton, W. A., Hirshfield, I. N., & Furtak, K Cell. 74, 909. (1993). https://doi.org/10.1016/0092-8674(93)90470-B
  13. Hartl, F. U. Nature. 381, 571. (1996). https://doi.org/10.1038/381571a0
  14. Hartl, F. U., & hayer-Hartl, M. Science. 295, 1852. (2002). https://doi.org/10.1126/science.1068408
  15. Fenton, W. A., & Horwich, A. L. Science. 6, 743. (1997).
  16. Brain, K., Otwinowdki, Z., Hegde, R., Boisvert, D. C., Joachimiak, A., Horwich, A. L., et al. Nature. 371, 578. (1994). https://doi.org/10.1038/371578a0
  17. Hunt, J. F., Weaver, A. J., Landry, S. J., Gierash, L., & Deisenhofer, J. Nature. 379, 37. (1996). https://doi.org/10.1038/379037a0
  18. Xu, Z., Horwich, A. L., & Sigler, P. B. Nature. 388, 741. (1997). https://doi.org/10.1038/41944
  19. Jonathan, S. W., Hays, S. R., Wayne, A. F., Joseph, M. B., Arthur, L. H. Cell Press. 84, 481. (1996).
  20. Garnier, J., Garnier, J. F., Robson, B., R. F. Doolittle, Editor. 266, 540. (1996). https://doi.org/10.1016/S0076-6879(96)66034-0
  21. Dosztanyi, Z., Csizmok, V., Tompa, P. and Simon, I. J. Mol. Biol. 347, 827. (2005). https://doi.org/10.1016/j.jmb.2005.01.071
  22. Delaglio, F., et al., J. Biomol. NMR. 6(3), 277. (1995).
  23. Johnson, B. A. and R. A. Blevins, J. Biomol. Nmr. 4(5), 603. (1994). https://doi.org/10.1007/BF00404272
  24. P. Goloubinoff, A. A. Gatenby, and G. H. Lorimer, Nature. 337, 44. (1989). https://doi.org/10.1038/337044a0
  25. Nishihara, K., Kanemori, M., Yanagi, H., & Yura, Y. Applied and Environmental Microbiology. 66, 884. (2000). https://doi.org/10.1128/AEM.66.3.884-889.2000
  26. Gasteiger, E., Hoogland, C., Gattiker, A., Duvaud, S., Wilkins, Marc R., Appel, Ron. D., Bairoch, A., J. M. Walker, Editor. Humana Press. 52,571.(2005).

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