Journal of Biomolecular NMR

, Volume 20, Issue 2, pp 135–147

Improved 3D triple resonance experiments, HNN and HN(C)N, for HN and 15N sequential correlations in (13C, 15N) labeled proteins: Application to unfolded proteins

  • Sanjay C. Panchal
  • Neel S. Bhavesh
  • Ramakrishna V. Hosur
Article

Abstract

Two triple resonance experiments, HNN and HN(C)N, are presented which correlate HN and 15N resonances sequentially along the polypeptide chain of a doubly (13C, 15N) labeled protein. These incorporate several improvements over the previously published sequences for a similar purpose and have several novel features. The spectral characteristics enable direct identification of certain triplets of residues, which provide many starting points for the sequential assignment procedure. The experiments are sensitive and their utility has been demonstrated with a 22 kDa protein under unfolding conditions where most of the standard triple resonance experiments such as HNCA, CBCANH etc. have limited success because of poor amide, Cα and Cβ chemical shift dispersions.

HN(C)N HNN sequential 15N correlations triple resonance experiments triplets unfolded proteins 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Bax, A. and Ikura, M. (1991) J. Biomol. NMR, 1, 99-104.Google Scholar
  2. Bax, A. and Grzesiek, S. (1993) Acc. Chem. Res., 26, 131-138.Google Scholar
  3. Booth, D.R., Sunde, M., Bellotti, V., Robinson, C.V., Hutchinson, W.L., Fraser, P.E., Hawkins, P.N., Dobson, C.M., Radford, S.E., Blake, C.C.F. and Pepys, M.B. (1997) Nature, 385, 787-793.Google Scholar
  4. Bracken, C., Palmer III, A.G. and Cavanagh, J. (1997) J. Biomol. NMR, 9, 94-100.Google Scholar
  5. Dill, K.A. and Shortle, D. (1991) Annu. Rev. Biochem., 60, 795-825.Google Scholar
  6. Grzesiek, S. and Bax, A. (1992a) J. Magn. Reson., 99, 201-207.Google Scholar
  7. Grzesiek, S. and Bax, A. (1992b) J. Am. Chem. Soc., 114, 6291-6293.Google Scholar
  8. Grzesiek, S., Anglister, J., Ren, H. and Bax, A. (1993) J. Am. Chem. Soc., 115, 4369-4370.Google Scholar
  9. Hennig, M., Bermel, W., Spencer, A., Dobson, C.M., Smith, L.J. and Schwalbe, H. (1999) J. Mol. Biol., 288, 705-723.Google Scholar
  10. Ikegami, T., Sato, S., Wälchli, M., Kyogoku, Y. and Shirakawa, M. (1997) J. Magn. Reson., 124, 214-217.Google Scholar
  11. Kay, L.E., Ikura, M., Tschudin, R. and Bax, A. (1990) J. Magn. Reson., 89, 496-514.Google Scholar
  12. Kay, L.E., Keifer, P. and Saarinen, T. (1992) J. Am. Chem. Soc., 114, 10663-10665.Google Scholar
  13. Kay, L.E. and Gardner, K.H. (1997) Curr. Opin. Struct. Biol., 7, 722-731.Google Scholar
  14. Kong, X.M., Kong, H.S. and Zhu, G. (1999) J. Biomol. NMR, 14, 133-140.Google Scholar
  15. Kupce, E., Matsuo, H. and Wagner, G. (1999) In Biological Magnetic Resonance: Modern Techniques in Protein NMR, Vol. 16 (Eds, Krishna, N.R. and Berliner, L.J.), Kluwer Academic/Plenum Publishers, Dordrecht, pp. 149-193.Google Scholar
  16. Lin, Y. and Wagner, G. (1999) J. Biomol. NMR, 15, 227-239.Google Scholar
  17. Liu, A., Riek, R., Wider, G., von Schroetter, C., Zahn, R. and Wüthrich, K. (2000) J. Biomol. NMR, 16, 127-138.Google Scholar
  18. Löhr, F., Pfeiffer, S., Lin, Y., Hartleib, J., Klimmek, O. and Rüterjans, H. (2000) J. Biomol. NMR, 18, 337-346.Google Scholar
  19. Logan, T.M., Theriault, Y. and Fesik, S.W. (1994) J. Mol. Biol., 236, 637-648.Google Scholar
  20. Matsuo, H., Kupce, E., Li, H. and Wagner, G. (1996) J. Magn. Reson., B111, 194-198.Google Scholar
  21. Muhandiram, D.R. and Kay, L.E. (1994) J. Magn. Reson., B103, 203-216.Google Scholar
  22. Penkett, C.J., Redfield, C., Jones, J.A., Dodd, J., Hubbard, J., Smith, R.A.G., Smith, L.J. and Dobson, C.M. (1998) Biochemistry, 37, 17054-17067.Google Scholar
  23. Pervushin, K., Riek, R., Wider, G. and Wüthrich, K. (1997) Proc. Natl. Acad. Sci. USA, 94, 12366-12371.Google Scholar
  24. Schwalbe, H., Fiebig, K.M., Buck, M., Jones, J.A., Grimshaw, S.B., Spencer, A., Glaser, S.J., Smith, L.J. and Dobson, C.M. (1997) Biochemistry, 36, 8977-8991.Google Scholar
  25. Shaka, A.J. (1985) Chem. Phys. Lett., 120, 201-205.Google Scholar
  26. Thomas, P.J., Qu, B.H. and Pederson, P.L. (1995) Trends Biochem. Sci., 20, 456-459.Google Scholar
  27. Weisemann, R., Rüterjans, H. and Bermel, W. (1993) J. Biomol. NMR, 3, 113-120.Google Scholar

Copyright information

© Kluwer Academic Publishers 2001

Authors and Affiliations

  • Sanjay C. Panchal
    • 1
  • Neel S. Bhavesh
    • 1
  • Ramakrishna V. Hosur
    • 1
  1. 1.Department of Chemical SciencesTata Institute of Fundamental ResearchMumbaiIndia

Personalised recommendations