Summary
We recently proposed a novel four-dimensional (4D) NMR strategy for the assignment of backbone nuclei in spectra of 13C/15N-labelled proteins (Boucher et al. (1992) J. Am. Chem. Soc., 114, 2262–2264 and J. Biomol. NMR, 2, 631–637). In this paper we extend this approach with a new constant time 4D HCC(CO)NNH experiment that also correlates the chemical shifts of the aliphatic sidechain (1H and 13C) and backbone (1H, 13Cα and 15N) nuclei. It separates the sidechain resonances, which may heavily overlap in spectra of proteins with large numbers of similar residues, according to the backbone nitrogen and amide proton chemical shifts. When used in conjunction with a 4D HCANNH or HNCAHA experiment it allows, in principle, complete assignment of aliphatic sidechain and backbone resonances with just two 4D NMR experiments.
Article PDF
Avoid common mistakes on your manuscript.
References
Bax, A., Clore, G.M., Driscoll, P.C., Gronenborn, A.M., Ikura, M. and Kay, L.E. (1990a) J. Magn. Reson., 87, 620–627.
Bax, A., Clore, G.M. and Gronenborn, A.M. (1990b) J. Magn. Reson., 88, 425–431.
Bax, A., Ikura, M., Kay, L.E. and Zhu, G. (1991) J. Magn. Reson., 91, 174–178.
Boucher, W., Raine, A.R.C. and Laue, E.D. (1991) NATO ASI series, 225, 87–103.
Boucher, W., Laue, E.D., Campbell-Burk, S.L. and Domaille, P.J. (1992a) J. Am. Chem. Soc., 114, 2262–2264.
Boucher, W., Laue, E.D., Campbell-Burk, S.L. and Domaille, P.J. (1992b) Biomol. NMR, 2, 631–637.
Burum, D.P. and Ernst, R.R. (1980) J. Magn. Reson., 39, 163–168.
Clore, G.M., Kay, L.E., Bax, A. and Gronenborn, A.M. (1991) Biochemistry, 30, 12–18.
Fesik, S.W., Eaton, H.L., Olejniczak, E.T., Zuiderweg, E.R.P., McIntosh, L.P. and Dahlquist, F.W. (1990) J. Am. Chem. Soc., 112, 886–888.
Grzesiek, S. and Bax, A. (1992) J. Am. Chem. Soc., 114, 6291–6293.
Hurd, R.E. (1990) J. Magn. Reson., 87, 422–428.
Ikura, M., Kay, L.E. and Bax, A. (1991a) J. Biomol. NMR, 1, 299–304.
Ikura, M., Spera, S., Barbato, G., Kay, L.E., Krinks, M. and Bax, A. (1991b) Biochemistry, 30, 9216–9228.
Johns, E.W. (1982) The HMG Chromosomal Proteins, Academic Press, London.
Kay, L.E., Clore, G.M., Bax, A. and Gronenborn, A.M. (1990a) Science, 249, 411–414.
Kay, L.E., Ikura, M., Tschudin, R. and Bax, A. (1990b) J. Magn. Reson., 89, 496–514.
Kay, L.E., Ikura, M., Grey, A.A. and Muhandiram, D.R. (1992a) J. Magn. Reson., 99, 652–659.
Kay, L.E., Wittekind, M., McCoy, M.A., Friedrichs, M.S. and Mueller, L. (1992b) J. Magn. Reson., 98, 443–450.
Laue, E.D., Mayger, M.R., Skilling, J. and Staunton, J. (1986) J. Magn. Reson., 68, 14–29.
Logan, T.M., Olejniczak, E.T., Xu, R.X. and Fesik, S.W. (1992) FEBS Lett., 314, 413–418.
Marion, D., Ikura, M. and Bax, A. (1989a) J. Magn. Reson., 84, 425–430.
Marion, D., Ikura, M., Tschudin, R. and Bax, A. (1989b) J. Magn. Reson., 85, 393–399.
Messerle, B.A., Wider, G., Otting, G., Weber, C. and Wüthrich, K. (1989) J. Magn. Reson., 85, 608–613.
Montelione, G.T., Lyons, B.A., Emerson, S.D. and Tashiro, M. (1992) J. Am. Chem. Soc., 114, 10974–10975.
Morris, G.A. and Freeman, R. (1979) J. Am. Chem. Soc., 101, 760–762.
Rance, M. (1987) J. Magn. Reson., 74, 557–564.
Spera, S. and Bax, A. (1991) J. Am. Chem. Soc., 113, 5490–5492.
Vuister, G.W. and Bax, A. (1992) J. Magn. Reson., 98, 428–435.
Wüthrich, K. (1986) NMR of Proteins and Nucleic Acids, Wiley, New York.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Clowes, R.T., Boucher, W., Hardman, C.H. et al. A 4D HCC(CO)NNH experiment for the correlation of aliphatic side-chain and backbone resonances in 13C/15N-labelled proteins. J Biomol NMR 3, 349–354 (1993). https://doi.org/10.1007/BF00212520
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00212520