Summary
The pulse sequence of a new constant-time 3D triple-resonance experiment, ct-HA[CAN]HN, is presented. This experiment delineates exclusively scalar connectivities and uses 13Cα−15N heteronuclear two-spin coherence to overlay the chemical shift evolution periods of the 13Cα and 15N nuclei, thereby providing the four resonance frequencies of the α-proton, the α-carbon, the amide nitrogen, and the amide proton of a given amino acid residue in three dimensions. This experiment promises to be a valid alternative to 4D experiments, providing the same information on intraresidue polypeptide backbone connectivities in 13C-15N-double-labeled proteins.
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Abbreviations
- 3D, 4D:
-
three-dimensional, four-dimensional
- TPPI:
-
time-proportional phase incrementation
- ct:
-
constant-time
- rf:
-
radiofrequency
- NOE:
-
nuclear Overhauser enhancement
- NOESY:
-
two-dimensional nuclear Overhauser enhancement spectroscopy
- glutaredoxin(C14S):
-
mutant E. coli glutaredoxin with the cysteine at position 14 replaced by serine
References
Bax, A., Ikura, M., Kay, L.E., Torchia, D.A. and Tschudin, R. (1990) J. Magn. Reson., 86, 304–318.
Bax, A. and Ikura, M. (1991) J. Biomol. NMR, 1, 99–104.
Boucher, W. and Laue, E.D. (1992) J. Am. Chem. Soc., 114, 2262–2264.
Bushweller, J.H., Åslund, F., Wüthrich, K. and Holmgren, A. (1992) Biochemistry, 31, 9288–9293.
Clubb, R.T., Thanabal, V. and Wagner, G. (1992) J. Biomol. NMR, 2, 203–210.
DeMarco, A. and Wüthrich, K. (1976) J. Magn. Reson., 24, 201–204.
Grzesiek, S. and Bax, A. (1992) J. Magn. Reson., 96, 432–440.
Güntert, P., Dötsch, V., Wider, G. and Wüthrich, K. (1992a) J. Biomol. NMR, 2, 619–629.
Güntert, P., Schäfer, N., Otting, G. and Wüthrich, K. (1992b) J. Magn. Res., in press.
Ikura, M., Bax, A. and Kay, L.E. (1990) Biochemistry, 29, 4659–4667.
Kay, L.E., Wittekind, M., McCoy, M.A., Friedrichs, M.S. and Mueller, L. (1992) J. Magn. Reson., 98, 443–450.
Marion, D., Ikura, M., Tschudin, R. and Bax, A. (1989) J. Magn. Reson., 85, 393–399.
Neri, D., Billeter, M., Wider, G. and Wüthrich, K. (1992) Science, 257, 1559–1563.
Otting, G. and Wüthrich, K. (1988) J. Magn. Reson., 76, 569–574.
Sodano, P., Bushweller, J.H., Björnberg, O., Holmgren, A., Billeter, M. and Wüthrich, K. (1991) J. Mol. Biol., 221, 1311–1324.
Shaka, A.J., Keeler, J., Frenkiel, T. and Freeman, R. (1983) J. Magn. Reson., 52, 335–338.
Sørensen, O.W., Eich, G.W., Levitt, M.H., Bodenhausen, G. and Ernst, R.R. (1983) Prog. Nucl. Magn. Reson. Spectrosc., 16, 163–192.
Torchia, D.A., Sparks, S.W. and Bax, A. (1989) Biochemistry, 28, 5509–5524.
Wüthrich, K. (1986) NMR of Proteins and Nucleic Acids, Wiley, New York.
Wüthrich, K., Wider, G., Wagner, G. and Braun, W. (1982) J. Mol. Biol., 155, 311–319.
Wüthrich, K., Spitzfaden, C., Memmert, K., Widmer, H. and Wider, G. (1991) FEBS Lett., 285, 237–247.
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Szyperski, T., Wider, G., Bushweller, J.H. et al. 3D 13C-15N-heteronuclear two-spin coherence spectroscopy for polypeptide backbone assignments in 13C-15N-double-labeled proteins. J Biomol NMR 3, 127–132 (1993). https://doi.org/10.1007/BF00242481
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DOI: https://doi.org/10.1007/BF00242481