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Automated NMR resonance assignment strategy for RNA via the phosphodiester backbone based on high-dimensional through-bond APSY experiments

Krähenbühl, Barbara ; El Bakkali, Issam ; Schmidt, Elena ; Güntert, Peter ; Wider, Gerhard

In: Journal of Biomolecular NMR, 2014, vol. 59, no. 2, p. 87-93

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    Titre
    • Automated NMR resonance assignment strategy for RNA via the phosphodiester backbone based on high-dimensional through-bond APSY experiments
    Auteur
    • Krähenbühl, Barbara. Institute of Molecular Biology and Biophysics, ETH Zurich, 8093, Zurich, Switzerland
    • El Bakkali, Issam. Institute of Molecular Biology and Biophysics, ETH Zurich, 8093, Zurich, Switzerland
    • Schmidt, Elena. Institute of Biophysical Chemistry, Goethe University Frankfurt am Main, 60438, Frankfurt am Main, Germany
    • Güntert, Peter. Institute of Biophysical Chemistry, Goethe University Frankfurt am Main, 60438, Frankfurt am Main, Germany
    • Wider, Gerhard. Institute of Molecular Biology and Biophysics, ETH Zurich, 8093, Zurich, Switzerland
    Type de document
    • Postprint
    Langue
    • Anglais
    Publié dans
    • Journal of Biomolecular NMR, 2014, vol. 59, no. 2, p. 87-93. Springer Netherlands
    Autre version électronique
    Identifiant OAI-PMH
    • oai:doc.rero.ch:325253
    Summary
    A fast, robust and reliable strategy for automated sequential resonance assignment for uniformly [13C, 15N]-labeled RNA via its phosphodiester backbone is presented. It is based on a series of high-dimensional through-bond APSY experiments: a 5D HCP-CCH COSY, a 4D H1′C1′CH TOCSY for ribose resonances, a 5D HCNCH for ribose-to-base connection, a 4D H6C6C5H5 TOCSY for pyrimidine resonances, and a 4D H8C8(C)C2H2 TOCSY for adenine resonances. The utilized pulse sequences are partially novel, and optimized to enable long evolution times in all dimensions. The highly precise APSY peak lists derived with these experiments could be used directly for reliable automated resonance assignment with the FLYA algorithm. This approach resulted in 98% assignment completeness for all 13C-1H, 15N1/9 and 31P resonances of a stem-loop with 14 nucleotides.