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Probing biomolecular dynamics and intercations by combining chemical synthesis and NMR spectroscopy

Authors
  • Mestdach, Emeline
Publication Date
Sep 26, 2022
Source
HAL-Descartes
Keywords
Language
English
License
Unknown
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Abstract

My thesis focused on the development of novel biomolecular NMR methodologies to characterise dynamics and interactions of biological macromolecules, by exploring novel horizons of biology, chemistry and NMR spectroscopy. My work first focused on micro-RNAs (miRNAs), key genetic regulators, capable to block messenger RNA (mRNA) translation. The let-7 miRNA and its targets were chemically synthesized to incorporate 19F probes, allowing to describe the interaction between let-7 and its target mRNAs by NMR. The second project was centered on a transfer RNA (tRNA), an essential component of the translation. Modulation of tRNA dynamic was probed using NMR 15N spin relaxation, in the context of tRNA maturation. Here, multiple-field relaxation rates, were measured and analysed to capture the fast dynamics of this system. The third study aimed at characterising a protein involved in the T5 bacteriophage infection of bacteria, and probe its interaction with its bacteria receptor protein. Resonance assignment of LLP the T5 protein involved in phage infection has been performed, as well as the 3D structure characterisation. Spin relaxation was used to probe LLP dynamics and titration experiments with the bacteria receptor allowed to reveal the binding interface. An additional project (not detailed) involved the dynamic study of an aquaporin, a membrane protein acting as water channel, by solid-state NMR at multiple fields. All these examples exemplify how NMR spectroscopy can probe complex dynamic processes in various contexts.

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