Dynamics of IDPs investigated using high-resolution relaxometry
| Autoři | |
|---|---|
| Rok publikování | 2024 |
| Druh | Vyžádané přednášky |
| Fakulta / Pracoviště MU | |
| Citace | |
| Popis | Introduction: Intrinsically disordered proteins (IDPs) are known for their extensive flexibility, which provides them with unique features necessary for their biological functions. Relaxation experiments are routinely used to probe the dynamics of proteins. Still, their information content is limited by the choice of magnetic field of NMR spectrometers used to conduct the experiments. Aims: We aimed to improve the description of IDP dynamics using the high-resolution relaxometry (HRR) technique. We also wanted to test the applicability of this method to challenging systems like IDPs, which are known for low chemical shift dispersion and high demands for high spectral resolution to obtain residue-specific information. Results: High-resolution relaxometry allowed us to measure relaxation at multiple fields using a single NMR spectrometer and to acquire data at very low magnetic fields. We were able to collect spectra even at fields as low as 0.1 T with sufficient sensitivity to extract reliable relaxation rates. While the sample was positioned at a low field for the relaxation delay, it was moved to the high-field center (at 14.1 T) for polarisation and detection. The HRR rates were measured using multidimensional spectra, providing the resolution required by the studied IDPs. The interpretation of the HRR rates requires consideration of cross-relaxation pathways active during the measurements. The analysis protocol was validated using measurements of accurate relaxation rates at low field (0.33 T) using a two-field NMR spectrometer. Conclusions: We showed that HRR is fully compatible with standard relaxation experiments used to study the motions of IDPs. The HRR data significantly improve the sampling of the spectral density functions and thus provide an opportunity to reveal new details of biomolecular motions. |
| Související projekty: |