Boosting the resolution of low-field 15N relaxation experiments on intrinsically disordered proteins with triple-resonance NMR
| Autoři | |
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| Rok publikování | 2020 |
| Druh | Článek v odborném periodiku |
| Časopis / Zdroj | Journal of biomolecular NMR |
| Fakulta / Pracoviště MU | |
| Citace | |
| www | https://link.springer.com/article/10.1007%2Fs10858-019-00298-6 |
| Doi | http://dx.doi.org/10.1007/s10858-019-00298-6 |
| Klíčová slova | Nuclear magnetic resonance; Relaxation; Dynamics; Intrinsically disordered proteins; High-resolution relaxometry; Non-uniform sampling |
| Popis | 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 15Namide 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 delta subunit of RNA polymerase from Bacillus subtilis, a protein with highly repetitive amino-acid sequence and very low dispersion of backbone chemical shifts. |
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