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High-dimensionality 13C direct-detected NMR experiments for the automatic assignment of intrinsically disordered proteins

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Abstract

We present three novel exclusively heteronuclear 5D 13C direct-detected NMR experiments, namely (HN-flipN)CONCACON, (HCA)CONCACON and (H)CACON(CA)CON, designed for easy sequence-specific resonance assignment of intrinsically disordered proteins (IDPs). The experiments proposed have been optimized to overcome the drawbacks which may dramatically complicate the characterization of IDPs by NMR, namely the small dispersion of chemical shifts and the fast exchange of the amide protons with the solvent. A fast and reliable automatic assignment of α-synuclein chemical shifts was obtained with the Tool for SMFT-based Assignment of Resonances (TSAR) program based on the information provided by these experiments.

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Acknowledgments

This work was supported in part by the EC 7th Framework program BioNMR (contract 261863), by the EC Marie Curie ITN program IDPbyNMR (contract 264257) and by grant number IP2012 062772, funded by Polish Ministry of Science and Higher Education for years 2013–2014. AZK thanks the Foundation for Polish Science for support with the START and the POMOST programs.

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Authors and Affiliations

  1. Bruker BioSpin GmbH, Silberstreifen, 76287, Rheinstetten, Germany

    Wolfgang Bermel

  2. CERM, University of Florence, Via Luigi Sacconi 6, 50019, Sesto Fiorentino, Florence, Italy

    Isabella C. Felli, Leonardo Gonnelli, Alessandro Piai & Roberta Pierattelli

  3. Department of Chemistry “Ugo Schiff”, University of Florence, 50019, Sesto Fiorentino, Italy

    Isabella C. Felli & Roberta Pierattelli

  4. Faculty of Chemistry, Biological and Chemical Research Centre, University of Warsaw, Żwirki i Wigury 101, 02-089, Warsaw, Poland

    Wiktor Koźmiński & Anna Zawadzka-Kazimierczuk

Authors
  1. Wolfgang Bermel
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  2. Isabella C. Felli
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  3. Leonardo Gonnelli
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  4. Wiktor Koźmiński
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  5. Alessandro Piai
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  6. Roberta Pierattelli
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  7. Anna Zawadzka-Kazimierczuk
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Correspondence to Isabella C. Felli or Roberta Pierattelli.

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Bermel, W., Felli, I.C., Gonnelli, L. et al. High-dimensionality 13C direct-detected NMR experiments for the automatic assignment of intrinsically disordered proteins. J Biomol NMR 57, 353–361 (2013). https://doi.org/10.1007/s10858-013-9793-z

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  • DOI: https://doi.org/10.1007/s10858-013-9793-z

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