Overbeck, Jan H. and Kremer, Werner and Sprangers, Remco (2020) A suite of F-19 based relaxation dispersion experiments to assess biomolecular motions. JOURNAL OF BIOMOLECULAR NMR, 74 (12). pp. 753-766. ISSN 0925-2738, 1573-5001
Full text not available from this repository. (Request a copy)Abstract
Proteins and nucleic acids are highly dynamic bio-molecules that can populate a variety of conformational states. NMR relaxation dispersion (RD) methods are uniquely suited to quantify the associated kinetic and thermodynamic parameters. Here, we present a consistent suite of F-19-based CPMG, on-resonance R-1 rho and off-resonance R-1 rho RD experiments. We validate these experiments by studying the unfolding transition of a 7.5 kDa cold shock protein. Furthermore we show that the F-19 RD experiments are applicable to very large molecular machines by quantifying dynamics in the 360 kDa half-proteasome. Our approach significantly extends the timescale of chemical exchange that can be studied with F-19 RD, adds robustness to the extraction of exchange parameters and can determine the absolute chemical shifts of excited states. Importantly, due to the simplicity of F-19 NMR spectra, it is possible to record complete datasets within hours on samples that are of very low costs. This makes the presented experiments ideally suited to complement static structural information from cryo-EM and X-ray crystallography with insights into functionally relevant motions.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | COLD-SHOCK PROTEIN; NUCLEAR-MAGNETIC-RESONANCE; MOLECULAR-WEIGHT PROTEINS; TIME-SCALE DYNAMICS; METHYL-GROUP PROBES; CHEMICAL-EXCHANGE; EXCITED-STATES; R-1-RHO RELAXATION; NMR-SPECTROSCOPY; CAVITY MUTANT; Fluorine; Large complexes; Protein folding; Relaxation dispersion; Structural dynamics |
| Subjects: | 500 Science > 570 Life sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Biophysik und physikalische Biochemie Biology, Preclinical Medicine > Institut für Biophysik und physikalische Biochemie > Prof. Dr. Remco Sprangers |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 10 Mar 2021 13:10 |
| Last Modified: | 10 Mar 2021 13:10 |
| URI: | https://pred.uni-regensburg.de/id/eprint/43736 |
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