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High resolution observed in 800 MHz DNP spectra of extremely rigid type III secretion needles

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Abstract

The cryogenic temperatures at which dynamic nuclear polarization (DNP) solid-state NMR experiments need to be carried out cause line-broadening, an effect that is especially detrimental for crowded protein spectra. By increasing the magnetic field strength from 600 to 800 MHz, the resolution of DNP spectra of type III secretion needles (T3SS) could be improved by 22 %, indicating that inhomogeneous broadening is not the dominant effect that limits the resolution of T3SS needles under DNP conditions. The outstanding spectral resolution of this system under DNP conditions can be attributed to its low overall flexibility.

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Acknowledgments

We thank Brigitta Angerstein for technical help, and Maximilian Zinke and Eve Ousby for valuable discussions. This work was supported by the Leibniz-Institut für Molekulare Pharmakologie, the Max Planck Society, the European Research Council (ERC Starting Grant to A.L.), the German Research Foundation (Deutsche Forschungsgemeinschaft; Emmy Noether Fellowship to A.L.) and the Fonds der Chemischen Industrie (Kekulé Scholarship to P.F.). Work at the 800 MHz DNP instrument at Utrecht was supported by NWO (Grants 700.11.344 and 700.58.102 to M.B.).

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Authors and Affiliations

  1. Department of Molecular Biophysics, Leibniz-Institut für Molekulare Pharmakologie, 13125, Berlin, Germany

    Pascal Fricke, Veniamin Chevelkov & Adam Lange

  2. NMR Research Group, Bijvoet Center for Biomolecular Research, Utrecht University, 3584 CH, Utrecht, The Netherlands

    Deni Mance & Marc Baldus

  3. Department of NMR-Based Structural Biology, Max Planck Institute for Biophysical Chemistry, 37077, Göttingen, Germany

    Karin Giller & Stefan Becker

  4. Institut für Biologie, Humboldt-Universität zu Berlin, 10115, Berlin, Germany

    Adam Lange

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  1. Pascal Fricke
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  2. Deni Mance
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  7. Adam Lange
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Correspondence to Adam Lange.

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Fricke, P., Mance, D., Chevelkov, V. et al. High resolution observed in 800 MHz DNP spectra of extremely rigid type III secretion needles. J Biomol NMR 65, 121–126 (2016). https://doi.org/10.1007/s10858-016-0044-y

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