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

Ioannis Gelis , Veronika Vitzthum (1) , Neha Dhimole , Marc A Caporini , Andreas Schedlbauer , Diego Carnevale , Sean R Connell , Paola Fucini , Geoffrey Bodenhausen (2)

1 ISIC - Institut des Sciences et Ingénierie Chimiques
2 UPMC - Université Pierre et Marie Curie - Paris 6

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.

Domaines: Chimie organique

Ioannis Gelis, Veronika Vitzthum, Neha Dhimole, Marc A Caporini, Andreas Schedlbauer, et al.. Solid-state NMR enhanced by dynamic nuclear polarization as a novel tool for ribosome structural biology.. Journal of Biomolecular NMR, 2013, 56 (2), pp.85-93. ⟨10.1007/s10858-013-9721-2⟩. ⟨hal-00825135⟩