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


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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Supplementary Figure (PDF 79 KB)


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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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