Skip to main content
SpringerLink
Log in

Measurement of residual dipolar couplings from 1Hα to 13Cα and 15N using a simple HNCA-based experiment

  • Published:
Journal of Biomolecular NMR Aims and scope Submit manuscript

Abstract

Novel NMR pulse schemes for simultaneous measurement of 1 D CαHαand 2 D NHαresidual dipolar couplings in proteins is presented. We show that 2 D NHαcoupling can be very useful for protein structure determination. The 2 D NHαcoupling can be measured from 15N dimension with good accuracy on a slowly relaxing TROSY resonance, utilizing HNCA-TROSY-based experiments, which concomitantly supply large 1 D CαHαcoupling. The dynamic range of 2 D NHαcoupling is comparable to 1 D NC′ coupling, but instead, it also serves non-redundant information on the course of protein backbone, thanks to rotational degree of freedom with respect to peptide bond. The HNCA-TROSY-based experiments are optimal for measuring residual dipolar couplings at high magnetic fields owing to absence of rapid transverse relaxation of carbonyl carbon. The reliability of the proposed approach was tested on 15N/13C human ubiquitin. A very good correlation with ubiquitin solution as well as crystal structure, for both 1 D CαHαand 2 D NHαcouplings, was obtained.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Annila, A., Aitio, H., Thulin, E. and Drakenberg, T. (1999) J. Biomol. NMR, 14, 223-230.

    Google Scholar 

  • Bax, A. and Tjandra, N. (1997) J. Biomol. NMR, 10, 289-292.

    Google Scholar 

  • Bax, A., Kontaxis, G. and Tjandra, N. (2001) Meth. Enzymol., 339, 127-174.

    Google Scholar 

  • Cai, M., Wang, H., Olejniczak, E.T., Meadows, R.P., Gunasekera, A.H., Xu, N. and Fesik, S. (1999) J. Magn. Reson., 139, 451-453.

    Google Scholar 

  • Carlomagno, T., Peti, W. and Griesinger, C. (2000) J. Biomol. NMR, 17, 99-109.

    Google Scholar 

  • Chou, J.J. and Bax, A. (2001) J. Am. Chem. Soc., 123, 3844-3845.

    Google Scholar 

  • Clore, G.M., Gronenborn, A.M. and Tjandra, N. (1998a) J. Magn. Reson., 131, 159-162.

    Google Scholar 

  • Clore, G.M., Gronenborn, A.M. and Bax, A. (1998b) J. Magn. Reson., 133, 216-221.

    Google Scholar 

  • Cornilescu, G., Marquardt, J.L., Ottiger, M. and Bax, A. (1998) J. Am. Chem. Soc., 120, 6836-6837.

    Google Scholar 

  • De Alba, E. and Tjandra, N. (2002) Prog. Nucl. Magn. Reson. Spectrosc., 40, 175-197.

    Google Scholar 

  • Hansen, M.R., Mueller, L. and Pardi, A. (1998) Nat. Struct. Biol., 5, 1065-1074.

    Google Scholar 

  • Kaikkonen, A. and Otting, G. (2001) J. Am. Chem. Soc., 123, 1770-1771.

    Google Scholar 

  • Kay, L.E., Keifer, P. and Saarinen, T. (1992) J. Am. Chem. Soc., 114, 10663-10665.

    Google Scholar 

  • Kontaxis, G. and Bax, A. (2001) J. Biomol. NMR, 20, 77-82.

    Google Scholar 

  • Logan, T.M., Olejniczak, E.T., Xu, X.R. and Fesik, S.W. (1993) J. Biomol. NMR, 3, 225-231.

    Google Scholar 

  • Loria, J.P., Rance, M. and Palmer III, A.G. (1999) J. Magn. Reson., 141, 180-184.

    Google Scholar 

  • Losonczi, J.A., Andrec, M., Fischer, M.W.F. and Prestegard, J.H. (1999) J. Magn. Reson., 138, 334-342.

    Google Scholar 

  • Marion, D., Ikura, M., Tschudin, R. and Bax, A. (1989) J. Magn. Reson., 85, 393-399.

    Google Scholar 

  • McCoy, M. and Mueller, L. (1992) J. Am. Chem. Soc., 114, 2108-2112.

    Google Scholar 

  • Meier, S., Hässinger, D., Jensen, P., Rogowski, M. and Grzesiek, S. (2003) J. Am. Chem. Soc., 125, 44-45.

    Google Scholar 

  • Meissner, A., Duus, J.Ø. and Sørensen, O.W. (1997) J. Biomol. NMR, 10, 89-94.

    Google Scholar 

  • Mittermaier, A. and Kay, L.E., (2001) J. Am. Chem. Soc., 123, 6892-6903.

    Google Scholar 

  • Ottiger, M., Delaglio, F. and Bax, A. (1998a) J. Magn. Reson., 131, 373-378.

    Google Scholar 

  • Ottiger, M., Delaglio, F., Marquardt, J.L., Tjandra, N. and Bax, A. (1998b) J. Magn. Reson., 134, 365-369.

    Google Scholar 

  • Otting, G., Rückert, M., Levitt, M.H. and Moshref, A. (2000) J. Biomol. NMR, 16, 343-346.

    Google Scholar 

  • Permi, P. (2001) J. Magn. Reson., 153, 267-272.

    Google Scholar 

  • Permi, P. (2002a) J. Biomol. NMR, 22, 27-35.

    Google Scholar 

  • Permi, P. (2002b) J. Biomol. NMR, 23, 201-209.

    Google Scholar 

  • Permi, P. and Annila, A. (2000) J. Biomol. NMR, 16, 221-227.

    Google Scholar 

  • Permi, P. and Annila, A. (2001b) J. Biomol. NMR, 20, 127-133.

    Google Scholar 

  • Permi, P. and Annila, A. (2002) J. Magn. Reson., 155, 123-130.

    Google Scholar 

  • Permi, P., Heikkinen, S, Kilpeläinen, I. and Annila, A. (1999) J. Magn. Reson., 139, 273-280.

    Google Scholar 

  • Permi, P., Rosevear, P.R. and Annila, A. (2000a) J. Biomol. NMR, 17, 43-54.

    Google Scholar 

  • Permi, P., Kilpeläinen, I. and Annila, A. (2000b) J. Magn. Reson., 146, 255-259.

    Google Scholar 

  • Pervushin, K., Riek, R., Wider, G. and Wüthrich, K. (1997) Proc. Natl. Acad. Sci. USA, 94, 12366-12371.

    Google Scholar 

  • Prestegard, J.H., Al-Hashimi, H.M. and Tolman, J.R. (2000) Quart. Rev. Biophys., 33, 371-424.

    Google Scholar 

  • Salzmann, M., Pervushin, K., Wider, G., Senn, H. and Wüthrich, K. (1998) Proc. Natl. Acad. Sci. USA, 15, 181-185.

    Google Scholar 

  • Seip, S., Balbach, J. and Kessler, H. (1992) Angew. Chem., 104, 1656-1658.

    Google Scholar 

  • Tian, F., Valafar, H. and Prestegard, J.H. (2001) J. Am. Chem. Soc., 123, 11791-11796.

    Google Scholar 

  • Tjandra, N. and Bax, A. (1997) Science, 278, 1111-1114.

    Google Scholar 

  • Tolman, J.R., Flanagan, J.M., Kennedy, M.A. and Prestegard, J.H. (1995) Proc. Natl. Acad. Sci. USA, 92, 9279-9283.

    Google Scholar 

  • Vijay-Kumar, S., Bugg, C.E. and Cook, W.J. (1987) J. Mol. Biol., 194, 531-544.

    Google Scholar 

  • Vögeli, B. and Pervushin, K. (2002) J. Biomol. NMR, 24, 291-300.

    Google Scholar 

  • Wand, A.C. and Bax, A. (1996) J. Am. Chem. Soc., 118, 2483-2494.

    Google Scholar 

  • Weigelt, J. (1998) J. Am. Chem. Soc., 120, 10778-10779.

    Google Scholar 

  • Wu, Z. and Bax, A. (2002) J. Am. Chem. Soc., 124, 9672-9673.

    Google Scholar 

  • Yang, D. and Nagayma, K. (1996) J. Magn. Reson., A118, 117-121.

    Google Scholar 

  • Yang, D. and Kay, L.E. (1999) J. Biomol. NMR, 14, 273-276.

    Google Scholar 

  • Yang, D., Tolman, J.R., Goto, N.K. and Kay, L.E. (1998) J. Biomol. NMR, 12, 325-332.

    Google Scholar 

  • Yang, D., Venters, R.A., Mueller, G.A., Choy, W.Y. and Kay, L.E. (1999) J. Biomol. NMR, 14, 333-343.

    Google Scholar 

  • Zweckstetter, M. and Bax, A. (2000) J. Am. Chem. Soc., 122, 3791-3792.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. NMR Laboratory, Structural Biology and Biophysics Programme, Institute of Biotechnology, University of Helsinki, P.O. Box 65, FIN-00014, Helsinki, Finland

    Perttu Permi

Authors
  1. Perttu Permi
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Perttu Permi.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Permi, P. Measurement of residual dipolar couplings from 1Hα to 13Cα and 15N using a simple HNCA-based experiment. J Biomol NMR 27, 341–349 (2003). https://doi.org/10.1023/A:1025866606252

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1025866606252

Search

Navigation