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Sequential assignments in uniformly 13C- and 15N-labelled RNAs: The HC(N,P) and HC(N,P)-CCH-TOCSY experiments

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Summary

An approach for the simultaneous acquisition of HCN and HCP as well as HCN-CCH-TOCSY and HCP-CCH-TOCSY triple resonance data sets for 13C-/15N-labelled RNAs is presented. The new HCN-CCH-TOCSY scheme unambiguously links all sugar resonances to the base nitrogen. In addition, simultaneous acquisition of HCN-CCH-TOCSY and HCP-CCH-TOCSY data sets provides sequential and base-type information in a single experiment, thereby saving data acquisition time as well as providing complementary data sets that are useful in clarifying ambiguous assignments. Virtually complete sequence-specific phosphate-ribose 1H, 31P, and base 15N1,9 assignments as well as partial 13C assignments could be obtained in a single experiment for a 0.5-mM sample of a 19-mer ribonucleotide.

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Authors and Affiliations

  1. Institut für Molekulare Biotechnologie, Postfach 100813, D-07708, Jena, Germany

    R. Ramachandran, C. Sich, M. Grüne, V. Soskic & L. R. Brown

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  1. R. Ramachandran
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  2. C. Sich
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  3. M. Grüne
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  4. V. Soskic
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  5. L. R. Brown
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Ramachandran, R., Sich, C., Grüne, M. et al. Sequential assignments in uniformly 13C- and 15N-labelled RNAs: The HC(N,P) and HC(N,P)-CCH-TOCSY experiments. J Biomol NMR 7, 251–255 (1996). https://doi.org/10.1007/BF00202042

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  • DOI: https://doi.org/10.1007/BF00202042

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