Skip to main content
Log in

Minimisation of sensitivity losses due to the use of gradient pulses in triple-resonance NMR of proteins

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

Summary

The use of pulsed field gradients in multiple-pulse NMR experiments has many advantages, including the possibility of obtaining excellent water suppression without the need for selective presaturation. In such gradient experiments the water magnetization is dephased deliberately; exchange between the saturated protons of the solvent water and the NH protons of a protein transfers this saturation to the protein. As the solvent is in large excess and relaxes relatively slowly, the result is a reduction in the sensitivity of the experiment due to the fact that the NH proton magnetization is only partially recovered. These effects can be avoided by ensuring that the water magnetization remains intact and is returned to the +z-axis at the start of data acquisition. General procedures for achieving this aim in any triple-resonance experiment are outlined and two specific examples are given. Experimental results confirm the sensitivity advantage of the modified sequences.

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

Access this article

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

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Bax, A., Sklenář, V., Clore, G.M. and Gronenborn, A.M. (1987) J. Am. Chem. Soc., 109, 6511–6513.

    Google Scholar 

  • Bax, A. and Pochapsky, S.S. (1992) J. Magn. Reson., 99, 638–643.

    Google Scholar 

  • Boucher, W., Laue, E.D., Campbell-Burk, S.L. and Domaille, P.J. (1992) J. Biomol. NMR, 2, 631–637.

    Google Scholar 

  • Cavanagh, J. and Rance, M. (1990) J. Magn. Reson., 88, 72–85.

    Google Scholar 

  • Clubb, R.T., Thanabal, V. and Wagner, G. (1992) J. Biomol. NMR, 2, 203–210.

    Google Scholar 

  • FarmerII, B.T., Venters, R.A., Spicer, L.D., Wittekind, M.G. and Müller, L. (1992) J. Biomol. NMR, 2, 195–202.

    Google Scholar 

  • Grzesiek, S. and Bax, A. (1992) J. Magn. Reson., 96, 432–440.

    Google Scholar 

  • Grzesiek, S. and Bax, A. (1993) J. Am. Chem. Soc., 115, 12593–12594.

    Google Scholar 

  • Ikura, M., Kay, L.E. and Bax, A. (1990) Biochemistry, 29, 4659–4667.

    Google Scholar 

  • Kay, L.E., Ikura, M. and Bax, A. (1990) J. Magn. Reson., 89, 496–514.

    Google Scholar 

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

    Google Scholar 

  • Kay, L.E., Wittekind, M., McCoy, M.A., Friedrichs, M.S. and Mueller, L. (1992b) J. Magn. Reson., 98, 443–450.

    Google Scholar 

  • Kay, L.E., Xu, G., Singer, A.U., Muhandiram, D.R. and Forman-Kay, J.D. (1993) J. Magn. Reson. Ser. B, 101, 333–337.

    Google Scholar 

  • Kay, L.E., Yi Xu, G. and Yamazaki, T. (1994) J. Magn. Reson. Ser. B, 109, 129–133.

    Google Scholar 

  • Kontaxis, G., Stonehouse, J., Laue, E.D. and Keeler, J. (1994) J. Magn. Reson. Ser. A, 111, 70–76.

    Google Scholar 

  • Li, Y. and Montelione, G.T. (1993) J. Magn. Reson. Ser. B, 101, 315–319.

    Google Scholar 

  • Marion, D. and Wüthrich, K. (1983) Biochem. Biophys. Res. Commun., 113, 967–974.

    Google Scholar 

  • Muhandiram, D.R. and Kay, L.E. (1994) J. Magn. Reson. Ser. B, 103, 203–216.

    Google Scholar 

  • Otting, G. and Wüthrich, K. (1988) J. Magn. Reson., 76, 569–574.

    Google Scholar 

  • Otting, G. (1994) J. Magn. Reson. Ser. B, 103, 288–291.

    Google Scholar 

  • PalmerIII, A.G., Cavanagh, J., Wright, P.E. and Rance, M. (1991) J. Magn. Reson., 93, 151–170.

    Google Scholar 

  • Shaka, A.J., Lee, C.J. and Pines, A. (1988) J. Magn. Reson., 77, 274–293.

    Google Scholar 

  • Sklenář, V., Tschudin, R. and Bax, A. (1987) J. Magn. Reson., 75, 352–357.

    Google Scholar 

  • Smallcombe, S.H. (1993) J. Am. Chem. Soc., 115, 4776–4785.

    Google Scholar 

  • States, D.J., Haberkorn, R.A. and Ruben, D.J. (1982) J. Magn. Reson., 48, 286–292.

    Google Scholar 

  • Stonehouse, J., Shaw, G.L., Keeler, J. and Laue, E.D. (1994a) J. Magn. Reson. Ser. A, 107, 178–184.

    Google Scholar 

  • Stonehouse, J., Shaw, G.L. and Keeler, J. (1994b) J. Biomol. NMR, 4, 799–805.

    Google Scholar 

  • Vuister, G.W., Clore, G.M., Gronenborn, A.M., Powers, R., Garrett, D.S., Tschudin, R. and Bax, A. (1993) J. Magn. Reson. Ser. B, 101, 210–213.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Stonehouse, J., Clowes, R.T., Shaw, G.L. et al. Minimisation of sensitivity losses due to the use of gradient pulses in triple-resonance NMR of proteins. J Biomol NMR 5, 226–232 (1995). https://doi.org/10.1007/BF00211750

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00211750

Keywords

Navigation