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

Authors

  • Ioannis Gelis
    • Buchmann Institute for Molecular Life Sciences, Institute of Organic Chemistry and Chemical BiologyGoethe University Frankfurt
    • Department of ChemistryUniversity of South Florida
  • Veronika Vitzthum
    • Institut des Sciences et Ingénierie ChimiquesEcole Polytechnique Fédérale de Lausanne
  • Neha Dhimole
    • Buchmann Institute for Molecular Life Sciences, Institute of Organic Chemistry and Chemical BiologyGoethe University Frankfurt
  • Marc A. Caporini
    • Institut des Sciences et Ingénierie ChimiquesEcole Polytechnique Fédérale de Lausanne
  • Andreas Schedlbauer
    • Buchmann Institute for Molecular Life Sciences, Institute of Organic Chemistry and Chemical BiologyGoethe University Frankfurt
    • Structural Biology Unit, CIC bioGUNEBizkaia Technology Park
  • Diego Carnevale
    • Institut des Sciences et Ingénierie ChimiquesEcole Polytechnique Fédérale de Lausanne
  • Sean R. Connell
    • Buchmann Institute for Molecular Life Sciences, Institute of Organic Chemistry and Chemical BiologyGoethe University Frankfurt
    • Structural Biology Unit, CIC bioGUNEBizkaia Technology Park
    • IKERBASQUEBasque Foundation for Science
    • Buchmann Institute for Molecular Life Sciences, Institute of Organic Chemistry and Chemical BiologyGoethe University Frankfurt
    • Structural Biology Unit, CIC bioGUNEBizkaia Technology Park
    • IKERBASQUEBasque Foundation for Science
    • Institut des Sciences et Ingénierie ChimiquesEcole Polytechnique Fédérale de Lausanne
    • Département de ChimieEcole Normale Supérieure
    • Université Pierre-et-Marie Curie
    • Laboratoire des Biomolécules, UMR 7203CNRS/UPMC/ENS
Article

DOI: 10.1007/s10858-013-9721-2

Cite this article as:
Gelis, I., Vitzthum, V., Dhimole, N. et al. J Biomol NMR (2013) 56: 85. doi:10.1007/s10858-013-9721-2

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