title: Solid-state NMR enhanced by dynamic nuclear polarization as a novel tool for ribosome structural biology.
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 (13)C and (15)N 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.
authors.affiliation: Buchmann Institute for Molecular Life Sciences, Institute of Organic Chemistry and Chemical Biology, Goethe University Frankfurt, Max-von-Laue-Str. 15, 60438 Frankfurt am Main, Germany.
authors.forename: Ioannis
authors.initials: I
authors.lastname: Gelis
authors.forename: Veronika
authors.initials: V
authors.lastname: Vitzthum
authors.forename: Neha
authors.initials: N
authors.lastname: Dhimole
authors.forename: Marc A
authors.initials: MA
authors.lastname: Caporini
authors.forename: Andreas
authors.initials: A
authors.lastname: Schedlbauer
authors.forename: Diego
authors.initials: D
authors.lastname: Carnevale
authors.forename: Sean R
authors.initials: SR
authors.lastname: Connell
authors.forename: Paola
authors.initials: P
authors.lastname: Fucini
authors.forename: Geoffrey
authors.initials: G
authors.lastname: Bodenhausen
mesh_terms: D005615:Freezing; D008958:Models, Molecular; D008968:Molecular Conformation; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods; D012270:Ribosomes / Q000737:chemistry*