Journal of Biomolecular NMR

, Volume 33, Issue 4, pp 199–211 | Cite as

SOFAST-HMQC Experiments for Recording Two-dimensional Deteronuclear Correlation Spectra of Proteins within a Few Seconds



Fast multidimensional NMR with a time resolution of a few seconds provides a new tool for high throughput screening and site-resolved real-time studies of kinetic molecular processes by NMR. Recently we have demonstrated the feasibility to record protein 1H–15N correlation spectra in a few seconds of acquisition time using a new SOFAST-HMQC experiment (Schanda and Brutscher (2005) J. Am. Chem. Soc. 127, 8014). Here, we investigate in detail the performance of SOFAST-HMQC to record 1H–15N and 1H−13C correlation spectra of proteins of different size and at different magnetic field strengths. Compared to standard 1H–15N correlation experiments SOFAST-HMQC provides a significant gain in sensitivity, especially for fast repetition rates. Guidelines are provided on how to set up SOFAST-HMQC experiments for a given protein sample. In addition, an alternative pulse scheme, IPAP-SOFAST-HMQC is presented that allows application on NMR spectrometers equipped with cryogenic probes, and fast measurement of one-bond 1H–13C and 1H–15N scalar and residual dipolar coupling constants.


Ernst angle fast NMR HMQC longitudinal relaxation enhancement protein residual dipolar couplings 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Andersson, P., Weigelt, J., Otting, G. 1998J. Biomol. NMR12435441CrossRefGoogle Scholar
  2. Atreya, H.S., Szyperski, T. 2004Proc. Natl. Acad. Sci. USA10196429647CrossRefGoogle Scholar
  3. Blackledge, M. 2005Prog. Nucl. Magn. Reson. Spectrosc.462361Google Scholar
  4. Brutscher, B. 2000Concept. Magnetic Res.12207229Google Scholar
  5. Brutscher, B. 2004J. Biomol. NMR295764CrossRefGoogle Scholar
  6. Brutscher, B., Simorre, J.P., Caffrey, M.S., Marion, D. 1994J. Magn. Reson. B1057782Google Scholar
  7. Emsley, L., Bodenhausen, G. 1992J. Magn. Reson.97135148Google Scholar
  8. Ernst, R., Bodenhausen, G., Wokaun, G. 1987Principles of Nuclear Magnetic Resonance in One and Two DimensionsOxford University PressOxfordGoogle Scholar
  9. Frydman, L., Scherf, T., Lupulescu, A. 2002Proc. Natl. Acad. Sci. USA991585815862CrossRefGoogle Scholar
  10. Geen, H., Freeman, R. 1991J. Magn. Reson.9393141Google Scholar
  11. Hajduk, P.J., Augeri, D.J., Mack, J., Mendoza, R., Yang, J.G., Betz, S.F., Fesik, S.W. 2000J. Am. Chem. Soc.12278987904CrossRefGoogle Scholar
  12. Hoch, J.C., Stern, A.S. 2001Nucl. Mag. Reson. Biol. Macromol., Pt A338159178Google Scholar
  13. Kim, S., Szyperski, T. 2003J. Am. Chem. Soc.12513851393Google Scholar
  14. Kupčce, E., Boyd, J., Campbell, I.D. 1995J. Magn. Reson. B106300303Google Scholar
  15. Kupče, E., Freeman, R. 1994J. Magn. Reson. A108268273Google Scholar
  16. Kupče, E., Freeman, R. 1996J. Magn. Reson. A118299303Google Scholar
  17. Kupče, E., Freeman, R. 2004J. Am. Chem. Soc.12664296440Google Scholar
  18. Kupče, E., Nishida, T., Freeman, R. 2003Prog. Nucl. Magn. Reson. Spectrosc.4295122Google Scholar
  19. Kupče, E., Wagner, G. 1995J. Magn. Reson. B109329333Google Scholar
  20. Mandelshtam, V.A. 2000J. Magn. Reson.144343356CrossRefGoogle Scholar
  21. Ottiger, M., Delaglio, F., Bax, A. 1998J. Magn. Reson.131373378CrossRefGoogle Scholar
  22. Pervushin, K., Vogeli, B., Eletsky, A. 2002J. Am. Chem. Soc.1241289812902CrossRefGoogle Scholar
  23. Piotto, M., Saudek, V., Sklenar, V. 1992J. Biomol. NMR2661665CrossRefGoogle Scholar
  24. Rosen, M.K., Gardner, K.H., Willis, R.C., Parris, W.E., Pawson, T., Kay, L.E. 1996J. Mol. Biol.263627636CrossRefGoogle Scholar
  25. Ross, A., Salzmann, M., Senn, H. 1997J. Biomol. NMR10389396CrossRefGoogle Scholar
  26. Ruckert, M., Otting, G. 2000J. Am. Chem. Soc.12277937797CrossRefGoogle Scholar
  27. Schanda, P., Brutscher, B. 2005J. Am. Chem. Soc.12780148015CrossRefGoogle Scholar
  28. Szyperski, T., Wider, G., Bushweller, J.H., Wuthrich, K. 1993J. Am. Chem. Soc.11593079308CrossRefGoogle Scholar
  29. Tjandra, N., Bax, A. 1997Science27811111114CrossRefGoogle Scholar
  30. Tugarinov, V., Hwang, P.M., Ollerenshaw, J.E., Kay, L.E. 2003J. Am. Chem. Soc.1251042010428Google Scholar
  31. Melckebeke, H., Simorre, J.P., Brutscher, B. 2004J. Am. Chem. Soc.12695849591Google Scholar
  32. Weigelt, J. 1998J. Am. Chem. Soc.1201077810779Google Scholar

Copyright information

© Springer 2005

Authors and Affiliations

  • Paul Schanda
    • 1
  • Ēriks Kupče
    • 2
  • Bernhard Brutscher
    • 1
  1. 1.Institut de Biologie Structurale – Jean-Pierre EbelUMR5075 CNRS-CEA-UJFGrenoble CedexFrance
  2. 2.Varian Ltd.Walton-on-ThamesSurreyUnited Kingdom

Personalised recommendations