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Boosting the resolution of low-field \(^{15}\hbox {N}\) relaxation experiments on intrinsically disordered proteins with triple-resonance NMR

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Abstract

Improving our understanding of nanosecond motions in disordered proteins requires the enhanced sampling of the spectral density function obtained from relaxation at low magnetic fields. High-resolution relaxometry and two-field NMR measurements of relaxation have, so far, only been based on the recording of one- or two-dimensional spectra, which provide insufficient resolution for challenging disordered proteins. Here, we introduce a 3D-HNCO-based two-field NMR experiment for measurements of protein backbone \(^{15}\hbox {N}\) amide longitudinal relaxation rates. The experiment provides accurate longitudinal relaxation rates at low field (0.33 T in our case) preserving the resolution and sensitivity typical for high-field NMR spectroscopy. Radiofrequency pulses applied on six different radiofrequency channels are used to manipulate the spin system at both fields. The experiment was demonstrated on the C-terminal domain of \(\delta\) subunit of RNA polymerase from Bacillus subtilis, a protein with highly repetitive amino-acid sequence and very low dispersion of backbone chemical shifts.

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Acknowledgements

This work was supported by Czech Science Foundation Grant Nos. GA18-04197Y and ANR-18-CE29-0003 provided by agence nationale de la recherche. Short scientific mission of PK to perform measurements at the two-field NMR spectrometer was supported by STSM Grant from the EURELAX COST Action CA15209.

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Author notes

  1. Z. Jaseňáková, V. Zapletal and P. Padrta contributed equally to this study.

Authors and Affiliations

  1. National Centre for Biomolecular Research, Faculty of Science and Central European Institute of Technology, Masaryk University, Kamenice 5, 625 00, Brno, Czech Republic

    Zuzana Jaseňáková, Vojtěch Zapletal & Lukáš Žídek

  2. Central European Institute of Technology, Masaryk University, Kamenice 5, 625 00, Brno, Czech Republic

    Petr Padrta & Pavel Kadeřávek

  3. Laboratoire des Biomolécules, LBM, Département de chimie, École normale supérieure, PSL University, Sorbonne Université, CNRS, 75005, Paris, France

    Milan Zachrdla, Nicolas Bolik-Coulon & Fabien Ferrage

  4. Bruker BioSpin GmbH, Silberstreifen 4, 76287, Rheinstetten, Germany

    Thorsten Marquardsen

  5. Bruker BioSpin, 34 rue de l’Industrie BP 10002, 67166, Wissembourg Cedex, France

    Jean-Max Tyburn

Authors
  1. Zuzana Jaseňáková
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  2. Vojtěch Zapletal
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Corresponding authors

Correspondence to Fabien Ferrage or Pavel Kadeřávek.

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Thorsten Marquardsen and Jean-Max Tyburn are employees of the Bruker BioSpin. The authors declare no other conflict of interest.

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Jaseňáková, Z., Zapletal, V., Padrta, P. et al. Boosting the resolution of low-field \(^{15}\hbox {N}\) relaxation experiments on intrinsically disordered proteins with triple-resonance NMR. J Biomol NMR 74, 139–145 (2020). https://doi.org/10.1007/s10858-019-00298-6

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