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Biochemistry (Moscow)

, Volume 74, Issue 1, pp 54–60 | Cite as

Isolation and crystallization of heterotrimeric translation initiation factor 2 from Sulfolobus solfataricus

  • E. A. StolboushkinaEmail author
  • O. S. Nikonov
  • M. B. Garber
Article

Abstract

The structure of the intact heterotrimeric translation initiation factor 2 (e/aIF2) is of great interest due to its key role in the initiator tRNA delivery to the ribosome and in translation initiation regulation in eukaryotes and archaea. We have chosen aIF2 from the hyperthermophilic archaeobacterium Sulfolobus solfataricus (SsoIF2) as an object for crystallization and structural investigations. Genes of the SsoIF2 subunits α, β, and γ were cloned and superexpressed. A method for heterotrimer SsoIF2αβγ purification was elaborated with at least 95% purity. Highly ordered crystals of the full-sized SsoIF2, reflecting X-rays at the resolution up to 2.8 Å, were obtained for the first time.

Key words

translation initiation e/aIF2 heterotrimer Sulfolobus solfataricus SsoIF2 crystals 

Abbreviations

DTT

1,4-dithiothreitol

e/aIF2

eukaryotic/archaean heterotrimeric translation initiation factor 2

PEG

polyethylene glycol

PEGMME

polyethylene glycol monomethyl ether

SsoIF2

aIF2 from the hyperthermophilic archaeobacterium Sulfolobus solfataricus

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References

  1. 1.
    Meunier, S., Spurio, R., Czisch, M., Wechselberger, R., Guenneugues, M., Gualerzi, C. O., and Boelens, R. (2000) EMBO J., 19, 1918–1926.PubMedCrossRefGoogle Scholar
  2. 2.
    Laursen, B. S., Mortensen, K. K., Sperling-Petersen, H. U., and Hoffman, D. W. (2003) J. Biol. Chem., 278, 16320–16328.PubMedCrossRefGoogle Scholar
  3. 3.
    Laursen, B. S., Kjaergaard, A. C., Mortensen, K. K., Hoffman, D. W., and Sperling-Petersen, H. U. (2004) Protein Sci., 13, 230–239.PubMedCrossRefGoogle Scholar
  4. 4.
    Wienk, H., Tomaselli, S., Bernard, C., Spurio, R., Picone, D., Gualerzi, C. O., and Boelens, R. (2005) Protein Sci., 14, 2461–2468.PubMedCrossRefGoogle Scholar
  5. 5.
    Kimball, S. R., Everson, W. V., Myers, L. M., and Jefferson, L. S. (1987) J. Biol. Chem., 262, 2220–2227.PubMedGoogle Scholar
  6. 6.
    Yatime, L., Schmitt, E., Blanquet, S., and Mechulam, Y. (2004) J. Biol. Chem., 279, 15984–15993.PubMedCrossRefGoogle Scholar
  7. 7.
    Pedulla, N., Palermo, R., Hasenohrl, D., Blasi, U., Cammarano, P., and Londei, P. (2005) Nucleic Acids Res., 33, 1804–1812.PubMedCrossRefGoogle Scholar
  8. 8.
    Das, A., Bagchi, M. K., Ghosh-Dastidar, P., and Gupta, N. K. (1982) J. Biol. Chem., 257, 1282–1288.PubMedGoogle Scholar
  9. 9.
    Erickson, F. L., and Hannig, E. M. (1996) EMBO J., 15, 6311–6320.PubMedGoogle Scholar
  10. 10.
    Colthurst, D. R., Campbell, D. G., and Proud, C. G. (1987) Eur. J. Biochem., 166, 357–363.PubMedCrossRefGoogle Scholar
  11. 11.
    Hinnebusch, A. G. (1996) in Translational Control of Gene Expression (Sonenberg, N., Hershey, J. W. B., and Mathews, M. B., eds.) Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, pp. 199–244.Google Scholar
  12. 12.
    Tahara, M., Ohsawa, A., Saito, S., and Kimura, M. (2004) J. Biochem. (Tokyo), 135, 479–485.Google Scholar
  13. 13.
    Thompson, G. M., Pacheco, E., Melo, E. O., and Castilho, B. A. (2000) J. Biochem., 347, 703–709.CrossRefGoogle Scholar
  14. 14.
    Cho, S., and Hoffman, D. W. (2002) Biochemistry, 41, 5730–5742.PubMedCrossRefGoogle Scholar
  15. 15.
    Das, S., Maiti, T., Das, K., and Maitra, U. (1997) J. Biol. Chem., 272, 31712–31718.PubMedCrossRefGoogle Scholar
  16. 16.
    Asano, K., Krishnamoorthy, T., Phan, L., Pavitt, G. D., and Hinnebusch, A. G. (1999) EMBO J., 18, 1673–1688.PubMedCrossRefGoogle Scholar
  17. 17.
    Das, S., and Maitra, U. (2000) Mol. Cell. Biol., 20, 3942–3950.PubMedCrossRefGoogle Scholar
  18. 18.
    Huang, H. K., Yoon, H., Hannig, E. M., and Donahue, T. F. (1997) Genes Dev., 11, 2396–2413.PubMedCrossRefGoogle Scholar
  19. 19.
    Schmitt, E., Blanquet, S., and Mechulam, Y. (2002) EMBO J., 21, 1821–1832.PubMedCrossRefGoogle Scholar
  20. 20.
    Nonato, M. C., Widom, J., and Clardy, J. (2002) J. Biol. Chem., 277, 17057–17061.PubMedCrossRefGoogle Scholar
  21. 21.
    Dhaliwal, S., and Hoffman, D. W. (2003) J. Mol. Biol., 334, 187–195.PubMedCrossRefGoogle Scholar
  22. 22.
    Roll-Mecak, A., Alone, P., Cao, C., Dever, T. E., and Burley, S. K. (2004) J. Biol. Chem., 279, 10634–10642.PubMedCrossRefGoogle Scholar
  23. 23.
    Ito, T., Marintchev, A., and Wagner, G. (2004) Structure, 12, 1693–1704.PubMedCrossRefGoogle Scholar
  24. 24.
    Gutierrez, P., Osborne, M. J., Siddiqui, N., Trempe, J. F., Arrowsmith, C., and Gehring, K. (2004) Protein Sci., 13, 659–667.PubMedCrossRefGoogle Scholar
  25. 25.
    Nikonov, O., Stolboushkina, E., Nikulin, A., Hasenohrl, D., Blasi, U., Manstein, D. J., Fedorov, D., Garber, M., and Nikonov, S. (2007) J. Mol. Biol., 373, 328–336.PubMedCrossRefGoogle Scholar
  26. 26.
    Yatime, L., Mechulam, Y., Blanquet, S., and Schmitt, E. (2006) Structure, 14, 119–128.PubMedCrossRefGoogle Scholar
  27. 27.
    Sokabe, M., Yao, M., Sakai, N., Toya, S., and Tanaka, I. (2006) Proc. Natl. Acad. Sci. USA, 103, 13016–13021.PubMedCrossRefGoogle Scholar
  28. 28.
    Yatime, L., Mechulam, Y., Blanquet, S., and Schmitt, E. (2007) Proc. Natl. Acad. Sci. USA, 104, 18445–18450.PubMedCrossRefGoogle Scholar
  29. 29.
    Studier, F. W., Rosenberg, A. H., Dunn, J. J., and Dubendorff, J. W. (1990) Meth. Enzymol., 185, 60–89.PubMedCrossRefGoogle Scholar
  30. 30.
    Laemmli, U. (1970) Nature, 227, 680–685.PubMedCrossRefGoogle Scholar
  31. 31.
    Davies, D., and Segal, D. (1971) Meth. Enzymol., 22, 266–269.CrossRefGoogle Scholar
  32. 32.
    Storoni, L. C., McCoy, A. J., and Read, R. J. (2004) Acta Crystallogr. Sect. D, 60, 432–438.CrossRefGoogle Scholar
  33. 33.
    Adams, P. D., Grosse-Kunstleve, R. W., Hung, L. W., Ioergew, T. R., McCoy, A. J., Moriarty, N. W., Read, R. J., Sacchettini, J. C., Sauter, N. K., and Terwilliger, T. C. (2002) Acta Crystallogr. Sect. D, 58, 1948–1954.CrossRefGoogle Scholar

Copyright information

© Pleiades Publishing, Ltd. 2009

Authors and Affiliations

  • E. A. Stolboushkina
    • 1
    Email author
  • O. S. Nikonov
    • 1
  • M. B. Garber
    • 1
  1. 1.Institute of Protein ResearchRussian Academy of SciencesPushchino Moscow RegionRussia

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