Journal of Biomolecular NMR

, Volume 63, Issue 1, pp 97–109

Dynamic nuclear polarization of nucleic acid with endogenously bound manganese

  • Patricia Wenk
  • Monu Kaushik
  • Diane Richter
  • Marc Vogel
  • Beatrix Suess
  • Björn Corzilius
Article

Abstract

We report the direct dynamic nuclear polarization (DNP) of 13C nuclei of a uniformly [13C,15N]-labeled, paramagnetic full-length hammerhead ribozyme (HHRz) complex with Mn2+ where the enhanced polarization is fully provided by the endogenously bound metal ion and no exogenous polarizing agent is added. A 13C enhancement factor of ε = 8 was observed by intra-complex DNP at 9.4 T. In contrast, “conventional” indirect and direct DNP experiments were performed using AMUPol as polarizing agent where we obtained a 1H enhancement factor of ε ≈ 250. Comparison with the diamagnetic (Mg2+) HHRz complex shows that the presence of Mn2+ only marginally influences the (DNP-enhanced) NMR properties of the RNA. Furthermore two-dimensional correlation spectra (15N–13C and 13C–13C) reveal structural inhomogeneity in the frozen, amorphous state indicating the coexistence of several conformational states. These demonstrations of intra-complex DNP using an endogenous metal ion as well as DNP-enhanced MAS NMR of RNA in general yield important information for the development of new methods in structural biology.

Keywords

Dynamic nuclear polarization (DNP) Solid-state NMR EPR RNA Transition metal Polarizing agent 

Supplementary material

10858_2015_9972_MOESM1_ESM.pdf (2.9 mb)
Supplementary material 1 (PDF 2987 kb)

Copyright information

© Springer Science+Business Media Dordrecht 2015

Authors and Affiliations

  • Patricia Wenk
    • 1
    • 3
  • Monu Kaushik
    • 1
  • Diane Richter
    • 1
  • Marc Vogel
    • 2
  • Beatrix Suess
    • 2
  • Björn Corzilius
    • 1
  1. 1.Institute of Physical und Theoretical Chemistry, Institute of Biophysical Chemistry und Center for Biomolecular Magnetic Resonance (BMRZ)Goethe UniversityFrankfurt am MainGermany
  2. 2.Department of BiologyTechnical University DarmstadtDarmstadtGermany
  3. 3.Werner Siemens Imaging Center and Department of Preclinical Imaging and RadiopharmacyUniversity of TübingenTübingenGermany

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