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Journal of Biomolecular NMR

, Volume 26, Issue 1, pp 79–83 | Cite as

Triple resonance experiments for the simultaneous correlation of H6/H5 and exchangeable protons of pyrimidine nucleotides in 13C,15N-labeled RNA applicable to larger RNA molecules

  • Jens WöhnertEmail author
  • Matthias Görlach
  • Harald Schwalbe
Article

Abstract

Triple-resonance two-dimensional H6/H5(C4N)H and C6/C5(C4N)H experiments are described that provide through-bond H6/H5 or C6/C5 to imino/amino correlations in pyrimidine bases in 13C,15N-labeled RNA. The experiments simultaneously transfer H6/H5 magnetization by an INEPT step to the C6/C5 nuclei and by homonuclear CC- and heteronuclear CN-TOCSY steps via the intervening C4 nucleus to the N3/N4 nuclei and then by a reverse INEPT step to the imino/amino hydrogens. The sensitivity of these experiments is high as demonstrated using a 30-nucleotide pyrimidine rich RNA at a concentration of 0.9 mM at temperatures of 10°C and 25°C. This indicates the general applicability of the experiments and the possibility to obtain correlations for imino resonances in non-canonical regions of the target RNA.

assignment 13C,15N-labeling NMR pyrimidine bases RNA TOCSY 

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Supplementary material

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References

  1. Fiala, R., Jiang, F. and Patel, D.J. (1996) J. Am. Chem. Soc., 118, 689-690.CrossRefGoogle Scholar
  2. Meiboom, S. and Gill, D. (1958) Rev. Sci. Instrum., 29, 688.CrossRefADSGoogle Scholar
  3. Mueller, L., Legault, P. and Pardi, A. (1995) J. Am. Chem. Soc., 117, 11043-11048.CrossRefGoogle Scholar
  4. Piotto, M., Saudek, V. and Sklenar, V. (1992) J. Biomol. NMR, 6, 661-665.CrossRefGoogle Scholar
  5. Shaka, A.J., Barker, P. and Freeman, R. (1985) J. Magn. Reson., 64, 547-557.Google Scholar
  6. Shaka, A.J., Lee, C.J. and Pines, A.J. (1988) J. Magn. Reson., 77, 274-293.Google Scholar
  7. Simorre, J.P., Zimmermann, A., Mueller, L. and Pardi, A. (1996a) J. Biomol. NMR, 7, 153-156.CrossRefGoogle Scholar
  8. Simorre, J.P., Zimmermann, A., Mueller, L. and Pardi, A. (1996b) J. Am. Chem. Soc., 118, 5316-5317.CrossRefGoogle Scholar
  9. Simorre, J.P., Zimmermann, A., Pardi, A., Farmer II, B.T. and Mueller, L. (1995) J. Biomol. NMR, 6, 427-432.CrossRefGoogle Scholar
  10. Sklenar, V., Diekmann, T., Butcher, S.E. and Feigon, J. (1996) J. Biomol. NMR, 7, 83-87.CrossRefGoogle Scholar
  11. States, D.J., Haberkorn, R.A. and Ruben, D.J. (1982) J. Magn. Reson., 48, 286-292.Google Scholar
  12. Stoldt, M., Wöhnert, J., Görlach, M. and Brown, L.R. (1998) EMBO J., 17, 6377-6384.CrossRefGoogle Scholar
  13. Varani, G., Aboul-Ela, F. and Allain, F.H. (1996) Prog. Nucl. Magn. Reson. Spectrosc., 29, 51-127.CrossRefGoogle Scholar
  14. Wijmenga, S.S. and van Buuren, B.N.M. (1998) Prog. Nucl. Magn. Reson. Spectrosc., 32, 287-387.CrossRefGoogle Scholar
  15. Wöhnert, J., Ramachandran, R., Görlach, M. and Brown, L.R. (1999) J. Magn. Reson., 139, 430-433.CrossRefADSGoogle Scholar

Copyright information

© Kluwer Academic Publishers 2003

Authors and Affiliations

  • Jens Wöhnert
    • 1
    Email author
  • Matthias Görlach
    • 2
  • Harald Schwalbe
    • 1
  1. 1.Institut für Organische Chemie und Chemische Biologie, Center for Biomolecular Magnetic ResonanceJohann Wolfgang Goethe-Universität FrankfurtFrankfurt/MGermany
  2. 2.Institut für Molekulare BiotechnologieJenaGermany

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