Abstract
A novel NMR experiment for obtaining sequential assignment of large proteins and protein complexes is described. The proposed method takes full advantage of transverse relaxation optimized spectroscopy (TROSY) and utilizes spin-state-selection to distinguish between intraresidual and sequential connectivities in the HNCA-TROSY-type correlation experiment. Thus, the intra- and interresidual cross peaks can be identified without relaying magnetization via carbonyl carbon, which relaxes very rapidly at the high magnetic fields where TROSY is most efficient. In addition, the presented method enables measurement of several scalar and residual dipolar couplings, which can potentially be used for structure determination of large proteins.
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Andersson, P., Weigelt, J. and Otting, G. (1998a) J. Biomol. NMR, 12, 435-441.
Andersson, P., Annila, A. and Otting, G. (1998b) J. Magn. Reson., 133, 364-367.
Astrof, N., Bracken, C., Cavanagh, J. and Palmer III, A.G. (1998) J. Biomol. NMR, 11, 451-456.
Gardner, K.H. and Kay, L.E. (1998) Annu. Rev. Biophys. Biomol. Struct., 27, 357-406.
Grzesiek, S. and Bax, A. (1993) J. Am. Chem. Soc., 115, 12593-12594.
Hu, J.S. and Bax, A. (1997) J. Am. Chem. Soc., 119, 6360-6368.
Konrat, R., Yang, D. and Kay, L.E. (1999) J. Biomol. NMR, 15, 309-313.
Loria, J.P., Rance, M. and Palmer III, A.G. (1999) J. Magn. Reson., 141, 180-184.
Marion, D., Ikura, M., Tschudin, R. and Bax, A. (1989) J. Magn. Reson., 85, 393-399.
Meissner, A., Duus, J.Ø. and Sø rensen, O.W. (1997) J. Biomol. NMR, 10, 89-94.
Meissner, A., Schulte-Herbrüggen, T. and Sø rensen, O.W. (1998a) J. Am. Chem. Soc., 120, 3803-3804.
Meissner, A., Schulte-Herbrüggen, T., Briand, J. and Sø rensen, O.W. (1998b) Mol. Phys., 95, 1137-1142.
Otomo, T., Teruya, K., Uegaki, K., Yamazaki, T. and Kyogoku, Y. (1999) J. Biomol. NMR, 14, 105-114.
Ottiger, M., Delaglio, F. and Bax, A. (1998) J. Magn. Reson., 131, 373-378.
Permi, P. and Annila, A. (2000) J. Biomol. NMR, 16, 221-227.
Permi, P., Sorsa, T., Kilpeläinen, I. and Annila, A. (1999a) J. Magn. Reson., 141, 44-51.
Permi, P., Heikkinen, S., Kilpeläinen, I. and Annila, A. (1999b) J. Magn. Reson., 140, 32-40.
Permi, P., Rosevear, P.R. and Annila, A. (2000) J. Biomol. NMR, 17, 43-54.
Pervushin, K., Riek, R., Wider, G. and Wüthrich, K. (1997) Proc. Natl. Acad. Sci. USA, 94, 12366-12371.
Pervushin, K.V., Wider, G. and Wüthrich, K. (1998) J. Biomol. NMR, 12, 345-348.
Rance, M., Loria, P. and Palmer III, A.G. (1999) J. Magn. Reson., 136, 92-101.
Salzmann, M., Pervushin, K., Wider, G., Senn, H. and Wüthrich, K. (2000) J. Am. Chem. Soc., 122, 7543-7548.
Shaka, A.J., Keeler, J., Frenkiel, T. and Freeman, R. (1983) J. Magn. Reson., 52, 335-338.
Shan, X., Gardner, K.H., Muhandiram, D.R., Rao, N.S., Arrowsmith, C.H. and Kay, L.E. (1996) J. Am. Chem. Soc., 118, 6570-6579.
Spezio, M., Wilson, D.B. and Karplus, P.A. (1993) Biochemistry, 32, 9906-9916.
Szyperski, T., Braun, D., Fernandez, C., Bartels, C. and Wüthrich, K. (1995) J. Magn. Reson., 108, 197-203.
Weigelt, J. (1998) J. Am. Chem. Soc., 120, 10778-10779.
Yamazaki, T., Lee, W., Revington, M., Mattiello, D.L., Dahlquist, F.W., Arrowsmith, C.H. and Kay, L.E. (1994) J. Am. Chem. Soc., 116, 6464-6465.
Yamazaki, T., Lee, W., Arrowsmith, C.H., Muhandiram, D.R. and Kay, L.E. (1994) J. Am. Chem. Soc., 116, 11655-11666.
Yamazaki, T., Otomo, T., Oda, N., Kyogoku, Y., Uegaki, K., Ito, N., Ishino, Y. and Nakamura, H. (1998) J. Am. Chem. Soc., 120, 5591-5592.
Yang, D. and Kay, L.E. (1999) J. Biomol. NMR, 13, 3-9.
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Permi, P., Annila, A. A new approach for obtaining sequential assignment of large proteins. J Biomol NMR 20, 127–133 (2001). https://doi.org/10.1023/A:1011208803036
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DOI: https://doi.org/10.1023/A:1011208803036