English

The effect of extrinsic paramagnetic probes on NMR relaxation rates for surface mapping of proteins and other biopolymers is a widely investigated and powerful NMR technique. Here we describe a new application of those probes. It relies on the setting of the relaxation delay to generate magnetization equilibrium and off-equilibrium conditions, in order to tailor the extent of steady state signal recovery with and without the water-soluble nitroxide Tempol. With this approach it is possible to identify signals whose relaxation is affected by exchange processes and, from the relative assignments, to map the protein residues involved in association or conformational interconversion processes on a micro-to-millisecond time scale. This finding is confirmed by the comparison with the results obtained from relaxation dispersion measurements. This simple and convenient method allows preliminary inspection to highlight regions where structural or chemical exchange events are operative, in order to focus on quantitative subsequent determinations by transverse relaxation dispersion experiments or analogous NMR relaxation studies, and/or to gain insights into the predictions of calculations. This presented approach can be advantageously applied to the characterization of the interface in protein-protein and protein-nanoparticles interactions and to study oligomerization of proteins.

Biophysics of protein conformational dynamics by NMR spectroscopy / Yamanappa Hunashal , 2021 Feb 25. 33. ciclo, Anno Accademico 2019/2020.

Biophysics of protein conformational dynamics by NMR spectroscopy

HUNASHAL, YAMANAPPA
2021-02-25

Abstract

English
25-feb-2021
The effect of extrinsic paramagnetic probes on NMR relaxation rates for surface mapping of proteins and other biopolymers is a widely investigated and powerful NMR technique. Here we describe a new application of those probes. It relies on the setting of the relaxation delay to generate magnetization equilibrium and off-equilibrium conditions, in order to tailor the extent of steady state signal recovery with and without the water-soluble nitroxide Tempol. With this approach it is possible to identify signals whose relaxation is affected by exchange processes and, from the relative assignments, to map the protein residues involved in association or conformational interconversion processes on a micro-to-millisecond time scale. This finding is confirmed by the comparison with the results obtained from relaxation dispersion measurements. This simple and convenient method allows preliminary inspection to highlight regions where structural or chemical exchange events are operative, in order to focus on quantitative subsequent determinations by transverse relaxation dispersion experiments or analogous NMR relaxation studies, and/or to gain insights into the predictions of calculations. This presented approach can be advantageously applied to the characterization of the interface in protein-protein and protein-nanoparticles interactions and to study oligomerization of proteins.
NMR; β2-microglobulin; D76N β2m; Gold nanoparticles
Biophysics of protein conformational dynamics by NMR spectroscopy / Yamanappa Hunashal , 2021 Feb 25. 33. ciclo, Anno Accademico 2019/2020.
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11390/1206968
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