Article

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 GelisAffiliated withBuchmann Institute for Molecular Life Sciences, Institute of Organic Chemistry and Chemical Biology, Goethe University FrankfurtDepartment of Chemistry, University of South Florida
  • , Veronika VitzthumAffiliated withInstitut des Sciences et Ingénierie Chimiques, Ecole Polytechnique Fédérale de Lausanne
  • , Neha DhimoleAffiliated withBuchmann Institute for Molecular Life Sciences, Institute of Organic Chemistry and Chemical Biology, Goethe University Frankfurt
  • , Marc A. CaporiniAffiliated withInstitut des Sciences et Ingénierie Chimiques, Ecole Polytechnique Fédérale de Lausanne
  • , Andreas SchedlbauerAffiliated withBuchmann Institute for Molecular Life Sciences, Institute of Organic Chemistry and Chemical Biology, Goethe University FrankfurtStructural Biology Unit, CIC bioGUNE, Bizkaia Technology Park
  • , Diego CarnevaleAffiliated withInstitut des Sciences et Ingénierie Chimiques, Ecole Polytechnique Fédérale de Lausanne
  • , Sean R. ConnellAffiliated withBuchmann Institute for Molecular Life Sciences, Institute of Organic Chemistry and Chemical Biology, Goethe University FrankfurtStructural Biology Unit, CIC bioGUNE, Bizkaia Technology ParkIKERBASQUE, Basque Foundation for Science
  • , Paola FuciniAffiliated withBuchmann Institute for Molecular Life Sciences, Institute of Organic Chemistry and Chemical Biology, Goethe University FrankfurtStructural Biology Unit, CIC bioGUNE, Bizkaia Technology ParkIKERBASQUE, Basque Foundation for Science Email author 
  • , Geoffrey BodenhausenAffiliated withInstitut des Sciences et Ingénierie Chimiques, Ecole Polytechnique Fédérale de LausanneDépartement de Chimie, Ecole Normale SupérieureUniversité Pierre-et-Marie CurieLaboratoire des Biomolécules, UMR 7203, CNRS/UPMC/ENS Email author 

Rent the article at a discount

Rent now

* Final gross prices may vary according to local VAT.

Get Access

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