Abstract
A pulse scheme resulting in improved sensitivity in TROSY-based 1HN-detected triple resonance experiments is presented. The approach minimizes relaxation losses which occur during the transfer of transverse magnetization from 15N to 1HN immediately prior to detection. The utility of the method is demonstrated on a complex of methyl protonated, highly deuterated maltose binding protein (MBP, 370 residues) and β- cyclodextrin. Sensitivity gains relative to previous TROSY schemes of approximately 10 and 20% are noted in HNCO spectra of MBP recorded at 25 and 5 °C, respectively, corresponding to molecular correlation times of 23 and 46 ns.
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Bax, A. (1994) Curr. Opin. Struct. Biol., 4, 738–744.
Boyd, J. and Scoffe, N. (1989) J. Magn. Reson., 85, 406–413.
Delaglio, F., Grzesiek, S., Vuister, G.W., Zhu, G., Pfeifer, J. and Bax, A. (1995) J. Biomol. NMR, 6, 277–293.
Farmer, B.T., Constantine, K.L., Goldfarb, V., Friedrichs, M.S., Wittekind, M., Yanchunas, J., Robertson, J.G. and Mueller, L. (1996) Nat. Struct. Biol., 3, 995–997.
Farrow, N.A., Muhandiram, R., Singer, A.U., Pascal, S.M., Kay, C.M., Gish, G., Shoelson, S.E., Pawson, T., Forman-Kay, J.D. and Kay, L.E. (1994) Biochemistry, 33, 5984–6003.
Gardner, K.H. and Kay, L.E. (1997) J. Am. Chem. Soc., 119, 7599–7600.
Gardner, K.H., Zhang, X., Gehring, K. and Kay, L.E. (1998) J. Am. Chem. Soc., 120, 11738–11748.
Garrett, D.S., Powers, R., Gronenborn, A.M. and Clore, G.M. (1991) J. Magn. Reson., 95, 214–220.
Garrett, D.S., Seok, Y.J., Liao, D.I., Peterkofsky, A., Gronenborn, A.M. and Clore, G.M. (1997) Biochemistry, 36, 2517–2530.
Kay, L.E. and Gardner, K.H. (1997) Curr. Opin. Struct. Biol., 7, 722–731.
Kay, L.E., Ikura, M., Tschudin, R. and Bax, A. (1990) J. Magn. Reson., 89, 496–514.
Kay, L.E., Keifer, P. and Saarinen, T. (1992) J. Am. Chem. Soc., 114, 10663–10665.
Kay, L.E., Xu, G.Y. and Yamazaki, T. (1994) J. Magn. Reson., A109, 129–133.
Marion, D., Ikura, M., Tschudin, R. and Bax, A. (1989) J. Magn. Reson., 85, 393–399.
Palmer, A.G., Cavanagh, J., Wright, P.E. and Rance, M. (1991) J. Magn. Reson., 93, 151–170.
Patt, S.L. (1992) J. Magn. Reson., 96, 94–102.
Pervushin, K., Riek, R., Wider, G. and Wüthrich, K. (1997) Proc. Natl. Acad. Sci. USA, 94, 12366–12371.
Pervushin, K., Riek, R., Wider, G. and Wüthrich, K. (1998a) J. Am. Chem. Soc., 120, 6394–6400.
Pervushin, K., Wider, G. and Wüthrich, K. (1998b) J. Biomol. NMR, 12, 345–348.
Schleucher, J., Sattler, M. and Griesinger, C. (1993) Angew. Chem. Int. Ed. Engl., 32, 1489–1491.
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.
Tessari, M., Vis, H., Boelens, R., Kaptein, R. and Vuister, G. (1997) J. Am. Chem. Soc., 119, 8985–8990.
Tjandra, N., Szabo, A. and Bax, A. (1996) J. Am. Chem. Soc., 118, 6986–6991.
Tjandra, N. and Bax, A. (1997) J. Am. Chem. Soc., 119, 8076–8082.
Venters, R.A., Farmer, B.T., Fierke, C.A. and Spicer, L.D. (1996) J. Mol. Biol., 264, 1101–1116.
Wüthrich, K. (1998) In Third European Conference on Stable Isotope Aided NMR of Biomolecules (Eds. Campbell, I., Laue, E. and Roberts, G.), St. John's College, Oxford.
Yu, L., Petros, A.M., Schnuchel, A., Zhong, P., Severin, J.M., Walter, K., Holzman, T.F. and Fesik, S.W. (1997) Nat. Struct. Biol., 4, 483–489.
Zhu, G. and Bax, A. (1990) J. Magn. Reson., 90, 405–410.
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Yang, D., Kay, L.E. Improved 1HN-detected triple resonance TROSY-based experiments. J Biomol NMR 13, 3–10 (1999). https://doi.org/10.1023/A:1008329230975
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DOI: https://doi.org/10.1023/A:1008329230975