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Automated NMR resonance assignment strategy for RNA via the phosphodiester backbone based on high-dimensional through-bond APSY experiments
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  • 作者:Barbara Kr?henbühl (1)
    Issam El Bakkali (1)
    Elena Schmidt (2)
    Peter Güntert (2)
    Gerhard Wider (1)
  • 关键词:Nucleic acids ; NMR ; Projection spectroscopy ; APSY ; Automated assignment ; FLYA ; Novel sampling methods
  • 刊名:Journal of Biomolecular NMR
  • 出版年:2014
  • 出版时间:June 2014
  • 年:2014
  • 卷:59
  • 期:2
  • 页码:87-93
  • 全文大小:
  • 参考文献:1. Aeschbacher T, Schubert M, Allain FH (2012) A procedure to validate and correct the 13C chemical shift calibration of RNA datasets. J Biomol NMR 52(2):179-90. doi:10.1007/s10858-011-9600-7 CrossRef
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  • 作者单位:Barbara Kr?henbühl (1)
    Issam El Bakkali (1)
    Elena Schmidt (2)
    Peter Güntert (2)
    Gerhard Wider (1)

    1. Institute of Molecular Biology and Biophysics, ETH Zurich, 8093, Zurich, Switzerland
    2. Institute of Biophysical Chemistry, Goethe University Frankfurt am Main, 60438, Frankfurt am Main, Germany
  • ISSN:1573-5001
文摘
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-sup class="a-plus-plus">1H, 15N1/9 and 31P resonances of a stem-loop with 14 nucleotides.

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