Boosting the resolution of low-field [Formula: see text] relaxation experiments on intrinsically disordered proteins with triple-resonance NMR.
Zuzana JaseňákováVojtěch ZapletalPetr PadrtaMilan ZachrdlaNicolas Bolik-CoulonThorsten MarquardsenJean-Max TyburnLukáš ŽídekFabien FerragePavel KadeřávekPublished in: Journal of biomolecular NMR (2020)
Improving our understanding of nanosecond motions in disordered proteins requires the enhanced sampling of the spectral density function obtained from relaxation at low magnetic fields. High-resolution relaxometry and two-field NMR measurements of relaxation have, so far, only been based on the recording of one- or two-dimensional spectra, which provide insufficient resolution for challenging disordered proteins. Here, we introduce a 3D-HNCO-based two-field NMR experiment for measurements of protein backbone [Formula: see text] amide longitudinal relaxation rates. The experiment provides accurate longitudinal relaxation rates at low field (0.33 T in our case) preserving the resolution and sensitivity typical for high-field NMR spectroscopy. Radiofrequency pulses applied on six different radiofrequency channels are used to manipulate the spin system at both fields. The experiment was demonstrated on the C-terminal domain of [Formula: see text] subunit of RNA polymerase from Bacillus subtilis, a protein with highly repetitive amino-acid sequence and very low dispersion of backbone chemical shifts.
Keyphrases
- single molecule
- high resolution
- amino acid
- magnetic resonance
- bacillus subtilis
- smoking cessation
- solid state
- human milk
- cross sectional
- optical coherence tomography
- magnetic resonance imaging
- ultrasound guided
- protein protein
- high frequency
- catheter ablation
- computed tomography
- atrial fibrillation
- ionic liquid
- quantum dots
- room temperature
- low birth weight
- contrast enhanced