Summary
Experiments and procedures are described that greatly alleviate the sequential assignment process of uniformly 13C/15N-enriched proteins by determining the type of amino acid from experiments that correlate side chain with backbone amide resonances. A recently proposed 3D NMR experiment, CBCA(CO)NH, correlates Cα and Cβ resonances to the backbone amide 1H and 15N resonances of the next residue (Grzesiek, S. and Bax, A. (1992) J. Am. Chem. Soc., 114, 6291–6293). An extension of this experiment is described which correlates the proton Hβ and Hα resonances to the amide 1H and 15N resonances of the next amino acid, and a detailed product operator description is given. A simple 2D-edited constant-time HSQC experiment is described which rapidly identifies Hβ and Cβ resonances of aromatic or Asn/Asp residues. The extent to which combined knowledge of the Cα and Cβ chemical shift values determines the amino acid type is investigated, and it is demonstrated that the combined Cα and Cβ chemical shifts of three or four adjacent residues usually are sufficient for defining a unique position in the protein sequence.
Article PDF
Similar content being viewed by others
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. and Grzesiek, S. (1993) Acc. Chem. Res., in press.
Boucher, W., Laue, E.D., Campbell-Burk, S. and Domaille, P.J. (1992) J. Am. Chem. Soc., 114, 2262–2264.
Burum, D.P. and Ernst, R.R. (1980) J. Magn. Reson., 39, 163–168.
Clore, G.M., Bax, A., Driscoll, P.C., Wingfield, P.T. and Gronenborn, A.M. (1990) Biochemistry, 29, 8172–8184.
Clubb, R.T., Thanabal, V. and Wagner, G. (1992) J. Magn. Reson., 97, 213–217.
Ernst, R.R., Bodenhausen, G. and Wokaun, A. (1987) Principles of Magnetic Resonance in One and Two Dimensions, Claredon Press, Oxford pp. 25–29.
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. (1992a) J. Magn. Reson., 96, 432–440.
Grzesiek, S. and Bax, A. (1992b) J. Magn. Reson., 99, 201–207.
Grzesiek, S. and Bax, A. (1992c) J. Am. Chem. Soc., 114, 6291–6293.
Grzesiek, S., Döbeli, H., Gentz, R., Garotta, G., Labhardt, A.M. and Bax, A. (1992) Biochemistry, 31, 8180–8190.
Hansen, P.E. (1991) Biochemistry, 30, 10457–10466.
Howarth, O.W. and Lilley, D.M.J. (1978) Proc. NMR Spectrosc., 12, 1–40.
Ikura, M., Kay, L.E. and Bax, A. (1990) Biochemistry, 29, 4659–4667.
Ikura, M., Kay, L.E., Krinks, M. and Bax, A. (1991) Biochemistry, 30, 5498–5504.
Ikura, M., Clore, G.M., Gronenborn, A.M., Zhu, G., Klee, C.B. and Bax, A. (1992) Science 256, 632–638.
Kay, L.E., Ikura, M., Tschudin, R. and Bax, A. (1990a) J. Magn. Reson. 89, 496–514.
Kay, L.E., Clore, G.M., Bax, A. and Gronenborn, A.M. (1990b) Science, 249, 411–414.
Kay, L.E., Ikura, M. and Bax, A. (1990c) J. Am. Chem. Soc., 112, 888–889.
Kay, L.E., Wittekind, M., McCoy, M.A., Friedrichs, M.S. and Mueller, L. (1992) J. Magn. Reson., 98, 443–450.
Marion, D., Driscoll, P.C., Kay, L.E., Wingfield, P.T., Bax, A., Gronenborn, A.M. and Clore, G.M. (1989a) Biochemistry, 28, 6150–6156.
Marion, D., Kay, L.E., Sparks, S.W., Torchia, D.A. and Bax, A. (1989b) J. Am. Chem. Soc., 111, 1515–1517.
Marion, D., Ikura, M., Tschudin, R. and Bax, A. (1989c) J. Mag. Res., 85, 393–399.
McCoy, M.A. and Mueller, L. (1992) J. Magn. Reson., 99, 18–36.
Meador, W.E., Means, A.R. and Quiocho, F.A. (1992) Science, 257, 1251–1255.
Oh, B.H., Westler, W.M., Darba, P. and Markley, J.L. (1988) Science, 240, 908–911.
Palmer III, A.G., Fairbrother, W.J., Cavanagh, J., Wright, P.E. and Rance, M. (1992) J. Biomol. NMR, 2, 103–108.
Pelton, J.G., Torchia, D.A., Meadow, N.D., Wong, C.-Y. and Roseman, S. (1991) Biochemistry, 30, 10043–10057.
Powers, R., Gronenborn, A.M., Clore, G.M. and Bax, A. (1991) J. Magn. Reson. 94, 209–213.
Santoro, J. and King, G.C. (1992) J. Magn. Reson. 97, 202–207.
Shaka, A.J., Lee, C.J. and Pines, A. (1988) J. Magn. Reson., 77, 274–293.
Spera, S. and Bax, A. (1991) J. Am. Chem. Soc., 113, 5490–5492.
van de Ven, F.J.M. and Philippens, M.E.P. (1992)J. Magn. Reson., 97, 637–644.
Vuister, G.W. and Bax, A. (1992) J. Magn. Reson., 98, 428–435.
Wagner, G. and Brühweiler, D. (1986), Biochemistry, 25, 5839–5843.
Wagner, G., Schneider, P. and Thanabal, V. (1991) J. Magn. Reson. 93, 436–440.
Wishart, D.S., Sykes, B.D. and Richards, F.M. (1991) J. Mol. Biol. 222, 311–333.
Wittekind, M., Görlach, M., Friedrichs, M., Dreyfuss, G. and Mueller, L. (1992) Biochemistry, 31, 6254–6265.
Wüthrich, K. (1986) NMR of Proteins and Nucleic Acids, Wiley, New York.
Zhu, G. and Bax, A. (1990) J. Magn. Reson., 90, 405–410.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Grzesiek, S., Bax, A. Amino acid type determination in the sequential assignment procedure of uniformly 13C/15N-enriched proteins. J Biomol NMR 3, 185–204 (1993). https://doi.org/10.1007/BF00178261
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00178261