期刊
JOURNAL OF BIOMOLECULAR NMR
卷 56, 期 2, 页码 85-93出版社
SPRINGER
DOI: 10.1007/s10858-013-9721-2
关键词
Ribosome structure; Magic-angle spinning NMR (MAS-NMR); Solid-state NMR; Dynamic nuclear polarization (DNP); TOTAPOL
资金
- Swiss National Science Foundation (SNSF)
- Swiss Commission for Technology and Innovation (CTI)
- Deutsche Forschungsgemeinschaft [FU579 1-3]
- Cluster of Excellence 'Macromolecular complexes' at the Goethe University Frankfurt (DFG) [EXC 115]
- Human Frontiers of Science Program Young Investigators Award [HFSP67/07]
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 C-13 and N-15 NMR correlation spectra can be recorded. Ribosome particles, directly pelleted and frozen into an NMR rotor, yield DNP signal enhancements on the order of similar to 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.
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