Abstract
NMR experiments are presented which allow backbone resonance assignment, secondary structure identification, and in favorable cases also molecular fold topology determination from a series of two-dimensional 1H-15N HSQC-like spectra. The 1H-15N correlation peaks are frequency shifted by an amount ± ωX along the 15N dimension, where ωX is the Cα, Cβ, or Hα frequency of the same or the preceding residue. Because of the low dimensionality (2D) of the experiments, high-resolution spectra are obtained in a short overall experimental time. The whole series of seven experiments can be performed in typically less than one day. This approach significantly reduces experimental time when compared to the standard 3D-based methods. The here presented methodology is thus especially appealing in the context of high-throughput NMR studies of protein structure, dynamics or molecular interfaces.
Similar content being viewed by others
References
Arnesano, F., Banci, L., Bertini, I., Ciofi-Baffoni, S., Molteni, E., Huffman, D.L. and O’Halloran, T.V. (2002) Genome Res., 12, 255-271.
Banci, L., Bertini, I., Ciofi-Baffoni, S., Huffman, D.L. and O’Halloran, T.V. (2001) J. Biol. Chem., 276, 8415-8426.
Bax, A., Kontaxis, G. and Tjandra, N. (2001) Meth. Enzymol., 339, 127-174.
Bodenhausen, G. and Ernst R.R. (1982) J. Am. Chem. Soc., 104, 1304-1309.
Brutscher, B., Morelle, N., Cordier, F. and Marion, D. (1995) J. Magn. Reson., B109, 238-242.
Brutscher, B., Simorre, J.-P., Caffrey, M. and Marion, D. (1994) J. Magn. Reson., B105, 77-82.
Brutscher, B. (2001) J. Magn. Reson., 151, 332-338.
Brutscher, B. (2002) J. Magn. Reson., 156, 155-159.
Ding, K. and Gronenborn, A. (2002) J. Magn. Reson., 156, 262-268.
Dosset, P., Hus J.-C. and Blackledge, M. (2001) J. Biomol. NMR, 20, 927-936.
Kim, S. and Szyperski, T. (2003) J. Am. Chem. Soc., 125, 1385-1393.
Montelione, G.T., Zhen, T., Huang, Y.J., Gunsalus, K.C. and Szyperski, T. (2000) Nat. Struct. Biol., 7, 982-984.
Morelle, N., Brutscher, B., Simorre, J.-P. and Marion, D. (1995) J. Biomol. NMR, 5, 154-160.
Nietlispach, D., Yutaka, I. and Laue, E. (2002) J. Am. Chem. Soc., 124, 11199-11207.
Ottiger, M., Delaglio, F. and Bax, A. J. Magn. Reson., 131, 373-378.
Palmer III, A.G., Cavanagh, J., Wright, P. and Rance, M. (1991) J. Magn. Reson., 93, 151-170.
Permi, P. (2002) J. Biomol. NMR, 23, 201-209.
Richarz, R. and Wüthrich, K. (1978) Biopolymers, 17, 2133-2141.
Rückert, M. and Otting, G. (2000) J. Am. Chem. Soc., 122, 7793-7797.
Simorre, J.-P., Brutscher, B., Caffrey, M. and Marion, D. (1994) J. Biomol. NMR, 4, 325-333.
Szyperski, T., Pellecchia, M. and Wüthrich, K. (1994) J. Magn. Reson., B105, 188-191.
Szyperski, T., Wider, G., Bushweller, J.H. and Wüthrich, K. (1993) J. Am. Chem. Soc., 115, 9307-9308.
Wang, A.C. and Bax, A. (1993) J. Biomol. NMR, 3, 715-720.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Bersch, B., Rossy, E., Covès, J. et al. Optimized set of two-dimensional experiments for fast sequential assignment, secondary structure determination, and backbone fold validation of 13C/15N-labelled proteins. J Biomol NMR 27, 57–67 (2003). https://doi.org/10.1023/A:1024746306675
Issue Date:
DOI: https://doi.org/10.1023/A:1024746306675