Description du sujet de thèse

This Ph.D. project aims at characterizing the structural mechanisms of a human receptor kinase protein involved in numerous diseases, such as cancers. The student will be involved in protein expression in bacteria and eukaryotic cells (insect and human cells), protein purification, structure determination (X-ray crystallography and cryo-EM), and biochemical characterization of protein function.

In our team, we are interested in a particular human receptor kinase that is involved in the regulation of cell growth. Its dysregulation leads to human diseases such as cancers, affecting millions of people worldwide. Therefore, this receptor is a very promising therapeutic target. However, the development of a new therapeutic solution targeting this receptor is limited by our current lack of understanding of its molecular mechanism, mainly due to the absence of structure for this receptor. The goal of the PhD student will be to solve the structure of the entire receptor or any of its isolated domains to shed light on its molecular mechanisms.

To this aim, the student will be involved in optimizing the recombinant protein expression system in eukaryotic cells. Different strategies for protein purification will be tested with several affinity tags, engineering protein constructs, and sampling orthologues. The student will purify soluble proteins and membrane proteins. Solubilization of membrane proteins will be performed in detergents or membrane-like systems (nanodiscs, amphipol). Protein samples will be screened for crystallization and cryogenic conditions to obtain atomic-resolution structures by X-Ray crystallography or cryo-EM methods. The structure obtained will be validated by functional assays in vitro (enzymology, ITC, SPR, TR-FRET, fluorescence polarization) and in cells (activity assay, binding assays, and cell proliferation assays).

Overall, the candidate will be trained to acquire vast expertise in integrative structural biology, biochemistry, biophysics, and cell biology.

Contexte de travail

This project will be carried out in collaboration with national and international academic laboratories whose objective is to study the molecular mechanisms of membrane receptors. New collaborations may be established during the development of the project.

Le poste se situe dans un secteur relevant de la protection du potentiel scientifique et technique (PPST), et nécessite donc, conformément à la réglementation, que votre arrivée soit autorisée par l'autorité compétente du MESR.