Applications

Solid-State DNP-NMR Spectrometer 263 GHz AVANCE™ III

The world’s first commercially available solid-state DNP-NMR spectrometer.

DNP-Enhanced Experiments on Large Biological Systems

Dynamic Nuclear Polarization (DNP) experiments at 263 GHz/ 400 MHz provide a large gain in sensitivity and dramatic reduction in signal averaging time. This enhanced sensitivity enables researchers to study samples that otherwise would have been inaccessible to NMR such as certain membrane proteins or samples which are available only in very limited amounts or dilute concentration. A range of samples have been successfully enhanced on the new 263 GHz Bruker DNP spectrometer including small peptides, soluble proteins, membrane proteins, and large biological complexes. The DNP samples are prepared by adding a polarizing agent (such as e.g. TOTAPOL biradical to a shared solvent or alternatively by utilizing a native radical on the sample of interest. The samples are measured under MAS while at low temperatures, typically 100–120 K. Unmodified NMR experiments are performed while benefitting from continuously DNP-enhanced signal intensity through CW microwave irradiation.

DNP-Enhanced CPMAS of 13C-Proline

DNP-Enhanced CPMAS of 13C-Proline in Glycerol/Water with 20 mM TOTAPOL:
25 μl sample, 1.5 mg U-13C-15N Proline, 8 kHz MAS, CPMAS with Spinal 64 decoupling, 110 k sample temperature, 8 seconds acquisition time for both microwaves on and off spectra

DNP-enhanced proton driven spin diffusion 13C-13C correlation experiment

DNP-enhanced proton driven spin diffusion 13C-13C correlation experiment on Barnase ribosome nascent chain complex with 10 mM TOTAPOL (nascent chain 13C labelled). 13 kHz MAS at 110 K. Experiment time: 17 hours.

Sample courtesy of Hartmut Oschkinat, Leibnizinstitut fuer Molekulare Pharmakologie.

DNP-enhanced NMR experiments on 15N Proteorhodopsin

DNP-enhanced NMR experiments on 15N Proteorhodopsin (WHYIF-reversely labeled) with 10 mM TOTAPOL. 6 mg of sample in 3.2 mm rotor. 8 kHz MAS at 105 K. Figure A: 15N CPMAS experiment showing single Schiff base resonance (SB) in 512 scans, 2 second recycle delay. Figure B: Twodimensional
15N-13C NCA correlation experiment to natural abundance 13C. Experiment time: 40 hours.

Sample courtesy of Vladimir Ladizhansky, University of Guelph.