Abstract
NMR spectra of ubiquitin in the presence of bicelles at a concentration of 25% w/v have been recorded under sample spinning conditions for different angles of rotation. For an axis of rotation equal to the magic angle, the 1H/15N HSQC recorded without any 1H decoupling in the indirect dimension corresponds to the classical spectrum obtained on a protein in an isotropic solution and allows the measurement of scalar J-couplings 1 J NH. For an angle of rotation smaller than the magic angle, the bicelles orient with their normal perpendicular to the spinning axis, whereas for an angle of rotation greater than the magic angle the bicelles orient with their normal along the spinning axis. This bicelle alignment creates anisotropic conditions that give rise to the observation of residual dipolar couplings in ubiquitin. The magnitude of these dipolar couplings depends directly on the angle that the rotor makes with the main magnetic field. By changing this angle in a controlled manner, residual dipolar couplings can be either scaled up or down thus offering the possibility to study simultaneously a wide range of dipolar couplings in the same sample.
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References
Barbara, T.M. (1994) J. Magn. Reson., 109, 265-269.
Bax, A. and Tjandra, N. (1997) J. Biomol. NMR, 10, 289-292.
Bax, A., Kontaxis, G. and Tjandra, N. (2001) Meth. Enzymol., 339, 127-174.
Clore, G.M., Starich, M.R. and Gronenborn, A.M. (1998) J. Am. Chem. Soc., 120, 10571-10572.
Cornilescu, G., Marquardt, J.L., Ottiger, M. and Bax, A. (1998) J. Am. Chem. Soc., 120, 6836-6837.
Courtieu, J., Alderman, D.W., Grant, D.M. and Bayles, J.P. (1982) J. Chem. Phys., 77, 723-730.
Courtieu, J., Bayle, J.P. and Fung, B.M. (1994) Prog. Nucl. Magn. Reson. Spectrosc., 26, 141-169.
Emsley, J.W. (1996) In Encyclopedia of Nuclear Magnetic Resonance, Vol. 4, Grant, D.M. and Harris, R.K. (eds.), Wiley, Chichester, pp. 2788-2799.
Emsley, J.W. (2002) In Solid-State NMR Spectroscopy Principles and Applications, Durer, M.J. (Ed.), Blackwell Science, Oxford, pp. 512-562.
Fung, B.M. (1996) In Encyclopedia of nuclear magnetic resonance, Vol. 4, Grant, D.M. and Harris, R.K. (Eds), Wiley, Chichester, pp. 2744-2751.
Glaubitz, C., Carravetta, M., Eden, M. and Levitt, M.H. (2001) In Perspectives on Solid State NMR, Kiihne, S.R. and de Groot, H.J.M. (Eds.), Kluwer Academic Publishers, Dordrecht, pp. 71-81.
Hansen, M.R., Mueller, L. and Pardi, A. (1998a) Nat. Struct. Biol., 5, 1065-1074.
Hansen, M.R., Rance, M. and Pardi, A. (1998b) J. Am. Chem. Soc., 120, 11210-11211.
Kay, L.E., Keifer, P. and Saarinen, T. (1992) J. Am. Chem. Soc., 114, 10663-10665.
Koenig, B.W., Hu, J.-S., Ottiger, M., Bose, S., Hendler, R.W. and Bax, A. (1999) J. Am. Chem. Soc., 121, 1385-1386.
Kooi, C.W.V., Kupce, E., Zuiderweg, E.R.P. and Pellecchia, M. (1999) J. Biomol. NMR, 15, 335-338.
Lipari, G. and Szabo, A. (1982) J. Am. Chem. Soc., 104, 4546-4559.
Lippens, G., Bourdonneau, M., Dhalluin, C., Warras, R., Richert, T., Seetharaman, C., Boutillon, C. and Piotto, M. (1999) Curr. Org. Chem., 3, 147-169.
Prestegard, J.H., Al-Hashimi, H.M. and Tolman, J.R. (2000) Q. Rev. Biophys., 33, 371-424.
Prestegard, J.H., Tolman, J.R., Al-Hashimi, H.M. and Andrec, M. (1999) In Biological Magnetic Resonance, Vol. 17, Krishna, N.R. and Berliner, L.J. (Eds.), Kluwer Academic Publishers, London, pp. 311-355.
Ruckert, M. and Otting, G. (2000) J. Am. Chem. Soc., 122, 7793-7797.
Sanders, C.R. and Schwonek, J.P. (1992) Biochemistry, 31, 8898-8905.
Sass, H.-J., Musco, G., Stahl, S.J., Wingfield, P.T. and Grzesiek, S. (2000) J. Biomol. NMR, 18, 303-309.
Sass, J., Cordier, F., Hoffmann, A., Rogowski, M., Cousin, A., Omichinski, J.G., Lowen, H. and Grzesiek, S. (1999) J. Am. Chem. Soc., 121, 2047-2055.
Saupe, A. (1968) Angew. Chem. Int. Ed. Engl., 7, 97.
Schmidt-Rohr, K. and Spiess, H.W. (1999) Multidimensional Solid-State NMR and Polymers, Academic Press, London.
Tian, F., Losonczi, J.A., Fischer, M.W.F. and Prestegard, J.H. (1999) J. Biomol. NMR, 15, 145-150.
Tjandra, N. and Bax, A. (1997) Science, 278, 1111-1114.
Tjandra, N., Grzesiek, S. and Bax, A. (1996a) J. Am. Chem. Soc., 118, 6264-6272.
Tjandra, N., Grzesiek, S. and Bax, A. (1996b) J. Am. Chem. Soc., 118, 9279-9287.
Tolman, J.R., Flanagan, J.M., Kennedy, M.A. and Prestegard, J.H. (1995) Proc. Natl. Acad. Sci. USA, 92, 9279.
Trempe, J.-F., Morin, F., Xia, P., Marchessault, R. and Gehring, K. (2002) J. Biomol. NMR, 22, 83-87.
Tycko, R., Blanco, F.J. and Ishii, Y. (2000) J. Am. Chem. Soc., 122, 9340-9341.
Zandomeneghi, G., Tomaselli, M., v. Beek, J.D. and Meier, B.H. (2001) J. Am. Chem. Soc., 123, 910-913.
Zandomeneghi, G., Tomaselli, M., Williamson, P.T.F. and Meier, B.H. (2003a) J. Biomol. NMR, 25, 113-123.
Zandomeneghi, G., Williamson, P.T.F., Hunkeler, A. and Meier, B.H. (2003b) J. Biomol. NMR, 25, 125-132.
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Lancelot, N., Elbayed, K., Bianco, A. et al. Measurement of Scaled Residual Dipolar Couplings in Proteins Using Variable-angle Sample Spinning. J Biomol NMR 29, 259–269 (2004). https://doi.org/10.1023/B:JNMR.0000032548.60663.1f
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DOI: https://doi.org/10.1023/B:JNMR.0000032548.60663.1f