Summary
A new four-dimensional pulse scheme is described for the main-chain assignment of proteins by means of the J connectivity of the amide proton and nitrogen resonances of adjacent residues. Since the new experiment, 4D CP-HN(COCA)NH, involves heteronuclear cross-polarization for magnetization transfer from 13C=O to 15N via 13Cα, a relatively strong WALTZ-16 decoupling rf field is applied to 13Cα during magnetization transfer. Consequently, 13Cα is effectively decoupled from its attached 2H in the case of deuterated proteins, in the absence of a decoupling rf field for 2H. This efficiently improves the sensitivity of the experiment through 13C line narrowing. The experiment was performed on a randomly 60% deuterated protein, and the sensitivity of the final 4D spectrum was found to be excellent.
Article PDF
Avoid common mistakes on your manuscript.
References
Abragam, A. (1961) The Principles of Nuclear Magnetism, Clarendon Press, Oxford.
Bax, A. and Grzesiek, S. (1993) Acc. Chem. Res. 26, 131–138.
Boucher, W., Raine, A.R.C. and Laue, E.D. (1991) In Computational Aspects of the Study of Biological Macromolecules by Nuclear Magnetic Resonance Spectroscopy (Eds, Hoch, J.C., Poulsen, F.M. and Redfield, C.) NATO ASI Series, Vol. 225, Plenum Press, New York, NY, p. 87.
Ernst, M., Griesinger, C. and Ernst, R.R. (1991) Mol. Phys., 74, 219–252.
Grzesiek, S., Anglister, J., Ren, H. and Bax, A. (1993) J. Am. Chem. Soc., 115, 4369–4370.
Ikura, M., Kay, L.E. and Bax, A. (1990) Biochemistry, 29, 4659–4667.
Kay, L.E., Ikura, M., Tschudin, R. and Bax, A. (1990) J. Magn. Reson., 89, 496–514.
Kushlan, D.M. and LeMaster, D.M. (1993) J. Biomol. NMR, 3, 701–708.
Laue, E.D., Mayger, M.R., Skilling, J. and Staunton, J. (1986) J. Magn. Reson., 68, 14–29.
Majumdar, A., Wang, H., Morshauser, R.C. and Zuiderweg, E.R.P. (1993) J. Biomol. NMR, 3, 387–397.
Marion, D., Ikura, M. and Bax, A. (1989) J. Magn. Reson. 84, 425–430.
Piotto, M., Saudek, V. and Sklenář, V., J. Biomol. NMR, 2, 661–665.
Richardson, J.M., Clowes, R.T., Boucher, W., Domaille, P.J., Hardman, C.H., Keeler, J. and Laue, E.D. (1993) J. Magn. Reson. Ser. B, 101, 223–227.
Schleucher, J., Schwendinger, M., Sattler, M., Schmidt, P., Glaser, S.J., Sørensen, O.W. and Griesinger, C. (1994) J. Biomol. NMR, 4, 301–306.
Vincent, S.J.F., Zwahlen, C. and Bodenhausen, G. (1993) J. Am. Chem. Soc., 115, 9202–9209.
Yamazaki, T., Lee, W., Revington, M., Mattiello, D., Dahlquist, F.W., Arrowsmith, C.H. and Kay, L.E. (1994) J. Am. Chem. Soc., 116, 6464–6465.
Zuiderweg, E.R.P. (1990) J. Magn. Reson., 89, 533–542.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Shirakawa, M., Wälchli, M., Shimizu, M. et al. The use of heteronuclear cross-polarization for backbone assignment of 2H-, 15N- and 13C-labeled proteins: A pulse scheme for triple-resonance 4D correlation of sequential amide protons and 15N. J Biomol NMR 5, 323–326 (1995). https://doi.org/10.1007/BF00211761
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00211761