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

• DOI: 10.1007/s10858-013-9721-2
• Publication Date: 2013-05-20T16:30:55Z
• Authors: Ioannis Gelis; Veronika Vitzthum; Neha Dhimole; Marc A. Caporini; Andreas Schedlbauer; Diego Carnevale; Sean R. Connell; Paola Fucini; Geoffrey Bodenhausen
• Keywords: Models, Molecular; 0301 basic medicine; 03 medical and health sciences; Freezing; Molecular Conformation; [CHIM.ORGA] Chemical Sciences/Organic chemistry; Nuclear Magnetic Resonance, Biomolecular; Ribosomes; 01 natural sciences; 0104 chemical sciences

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.