Abstract
A suite of multiple quantum (MQ) HCN-based pulse sequences has been developed for the purpose of collecting dipolar coupling data in labeled nucleic acids. All the pulse sequences are based on the robust MQ-HCN experiment which has been utilized for assignment purposes in labeled nucleic acids for a number of years and provides much-needed resolution for the dipolar coupling measurements. We have attempted to collect multiple couplings centered on the 13C1′ and 13C6/8 positions. Six pulse sequences are described, one each for measurement of one-bond 13C1′-1H1′ and 13C6/8-1H6/8 couplings, one for measurement of one-bond 13C1′-15N and two-bond 1H1′-15N couplings, one for measurement of one-bond13C6/8-15N and two-bond 1H6/8-15N couplings, one for measurement of one-bond 13C1′-13C2′ and two-bond 1H1′-13C2′ couplings, and one for measurement of one-bond 13C6-13C5 and two-bond 1H6-13C5 couplings in the bases of C and T. These sequences are demonstrated for a labeled 18 bp DNA duplex in a 47 kDa ternary complex of DNA, CBFβ, and the CBFα Runt domain, thus clearly demonstrating the robustness of the pulse sequences even for a very large complex.
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References
Bartels, C., Xia, T., Billeter, M., Guntert, P. and Wuthrich, K. (1995) J.Biomol. NMR, 6, 1–10.
Bayer, P., Varani, L. and Varani, G. (1999) J.Biomol. NMR, 14, 149–155.
Berardi, M., Sun, C., Zehr, M., Abildgaard, F., Peng, J., Speck, N.A. and Bushweller, J.H. (1999) Structure, 7, 1247–1256.
Brutscher, B., Boisbouvier, J., Pardi, A., Marion, D. and Simorre, J. (1998) J. Am. Chem. Soc.,120, 11845–11851.
Chen, X., Mariappan, S.V.S., Kelley, J.J., Bushweller, J.H., Bradbury, E.M. and Gupta, G. (1998) FEBS Lett., 436,372–376.
Farmer, B.T., Muller, L., Nikonowicz, E.P. and Pardi, A. (1993) J. Am. Chem. Soc., 115, 11040–11041.
Farmer, B.T., Muller, L., Nikonowicz, E.P. and Pardi, A. (1994) J. Biomol. NMR,4,129–133.
Fiala, R., Czernek, J. and Sklenar, V. (2000) J. Biomol. NMR,6, 291–302. 20
Fiala, R., Jiang, F. and Sklenar, V. (1998) J. Biomol. NMR, 12, 373–383.
Guntert, P., Dotsch, V., Wider, G. and Wuthrich, K. (1992) J. Biomol. NMR,2,619–629.
Kupce, E., Boyd, J. and Campbell, I.D. (1995) J. Magn. Reson., 106B,300.
Marino, J.P., Diener, J.L., Moore, P.B. and Griesinger, C. (1997) J. Am. Chem. Soc., 119,7361–7366.
Pervushin, K., Riek, R., Wider, G. and Wuthrich, K. (1998) J. Am. Chem. Soc.,120,6394–6400.
Sklenar, V., Dieckmann, T., Butcher, S.E. and Feigon, J. (1998) J. Magn. Reson.,130, 119–122.
Sklenar, V., Peterson, R.D., Rejante, M.R., Wang, E. and Feigon, J. (1993a) J. Am. Chem. Soc.,115, 12181–12182.
Sklenar, V., Peterson, R.D., Rejante, M.R. and Feigon, J. (1993b) J. Biomol. NMR,3,721–727.
Smallcombe, S.H., Patt, S.H. and Keifer, P.A. (1995) J. Magn. Reson., A117,295–303.
Tate, S., Ono, A. and Kainosho, M. (1994) J. Am. Chem. Soc., 116,5977–5978.
Tjandra, N. and Bax, A. (1997) J. Am. Chem. Soc.,119,8076–8082.
Tjandra, N., Tate, S.-I., Ono, A., Kainosho, M. and Bax, A. (2000) Science,278,1111–1114.
Yan, J. and Bushweller, J.H. (2001) Biochem. Biophys. Res. Commun., 284,295–300.
Yang, D., Venters, R. A., Mueller, G. A., Choy, W.Y. and Kay, L.E. (1999) J. Biomol. NMR,14, 333–343.
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Yan, J., Corpora, T., Pradhan, P. et al. MQ-HCN-based pulse sequences for the measurement of 13C1′-1H1′, 13C1′-15N, 1H1′-15N, 13C1′-13C2′, 1H1′-13C2′, 13C6/8-1H6/8, 13C6/8-15N, 1H6/8-15N, 13C6-13C5, 1H6-13C5 dipolar couplings in 13C, 15N-labeled DNA (and RNA). J Biomol NMR 22, 9–20 (2002). https://doi.org/10.1023/A:1013876105589
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DOI: https://doi.org/10.1023/A:1013876105589