Journal of Biomolecular NMR

, Volume 56, Issue 2, pp 85–93

Solid-state NMR enhanced by dynamic nuclear polarization as a novel tool for ribosome structural biology

  • Ioannis Gelis
  • Veronika Vitzthum
  • Neha Dhimole
  • Marc A. Caporini
  • Andreas Schedlbauer
  • Diego Carnevale
  • Sean R. Connell
  • Paola Fucini
  • Geoffrey Bodenhausen
Article

Abstract

The impact of Nuclear Magnetic Resonance (NMR) on studies of large macromolecular complexes hinges on improvements in sensitivity and resolution. Dynamic nuclear polarization (DNP) in the solid state can offer improved sensitivity, provided sample preparation is optimized to preserve spectral resolution. For a few nanomoles of intact ribosomes and an 800 kDa ribosomal complex we demonstrate that the combination of DNP and magic-angle spinning NMR (MAS-NMR) allows one to overcome current sensitivity limitations so that homo- and heteronuclear 13C and 15N NMR correlation spectra can be recorded. Ribosome particles, directly pelleted and frozen into an NMR rotor, yield DNP signal enhancements on the order of ~25-fold and spectra that exhibit narrow linewidths, suitable for obtaining site-specific information. We anticipate that the same approach is applicable to other high molecular weight complexes.

Keywords

Ribosome structure Magic-angle spinning NMR (MAS-NMR) Solid-state NMR Dynamic nuclear polarization (DNP) TOTAPOL 

Supplementary material

10858_2013_9721_MOESM1_ESM.pdf (1.3 mb)
Supplementary material 1 (PDF 1363 kb)

Copyright information

© Springer Science+Business Media Dordrecht 2013

Authors and Affiliations

  • Ioannis Gelis
    • 1
    • 2
  • Veronika Vitzthum
    • 3
  • Neha Dhimole
    • 1
  • Marc A. Caporini
    • 3
  • Andreas Schedlbauer
    • 1
    • 5
  • Diego Carnevale
    • 3
  • Sean R. Connell
    • 1
    • 5
    • 6
  • Paola Fucini
    • 1
    • 5
    • 6
  • Geoffrey Bodenhausen
    • 3
    • 4
    • 7
    • 8
  1. 1.Buchmann Institute for Molecular Life Sciences, Institute of Organic Chemistry and Chemical BiologyGoethe University FrankfurtFrankfurt am MainGermany
  2. 2.Department of ChemistryUniversity of South FloridaTampaUSA
  3. 3.Institut des Sciences et Ingénierie ChimiquesEcole Polytechnique Fédérale de LausanneLausanneSwitzerland
  4. 4.Département de ChimieEcole Normale SupérieureParis Cedex 05France
  5. 5.Structural Biology Unit, CIC bioGUNEBizkaia Technology ParkDerioSpain
  6. 6.IKERBASQUEBasque Foundation for ScienceBilbaoSpain
  7. 7.Université Pierre-et-Marie CurieParisFrance
  8. 8.Laboratoire des Biomolécules, UMR 7203CNRS/UPMC/ENSParisFrance

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