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Journal of Biomolecular NMR

, Volume 65, Issue 3–4, pp 121–126 | Cite as

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

  • Pascal Fricke
  • Deni Mance
  • Veniamin Chevelkov
  • Karin Giller
  • Stefan Becker
  • Marc Baldus
  • Adam LangeEmail author
Communication

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.

Keywords

Proteins Type III Secretion System Solid-State NMR DNP Dynamics PRE 

Notes

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.).

Supplementary material

10858_2016_44_MOESM1_ESM.pdf (557 kb)
Supplementary material 1 (PDF 557 kb)

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Copyright information

© Springer Science+Business Media Dordrecht 2016

Authors and Affiliations

  • Pascal Fricke
    • 1
  • Deni Mance
    • 2
  • Veniamin Chevelkov
    • 1
  • Karin Giller
    • 3
  • Stefan Becker
    • 3
  • Marc Baldus
    • 2
  • Adam Lange
    • 1
    • 4
    Email author
  1. 1.Department of Molecular BiophysicsLeibniz-Institut für Molekulare PharmakologieBerlinGermany
  2. 2.NMR Research Group, Bijvoet Center for Biomolecular ResearchUtrecht UniversityUtrechtThe Netherlands
  3. 3.Department of NMR-Based Structural BiologyMax Planck Institute for Biophysical ChemistryGöttingenGermany
  4. 4.Institut für BiologieHumboldt-Universität zu BerlinBerlinGermany

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