Missions

Bring more insights into the precise enzyme regulation of the NAD kinases and especially the human cytoplasmic one.

Activités

1. Prepare variants of the NAD kinases under study by means of directed mutagenesis, over-expression, purification and biophysical characterization.
2. Determine liganded structures of the variants and characterize their enzymatic activity.
3. Suggest optimization of the current inhibitors and evaluate the potency of new compounds.
4. Collaborate with biologists on the activation mechanism of NADK kinases in cells.

Compétences

These studies will rely on various techniques including:

- Molecular Biology (plasmid constructions) including directed mutagenesis.
- Protein production in bacteria and eukaryotic cells.
- Protein-protein interaction assays (i.e: fluorescence anisotropy, ITC).
- Purification of proteins suitable for cryoEM analysis, SAXS and X-ray crystallography.
- Preparation and visualization of cryoEM grids.
- CryoEM data treatment.
- sample preparation for mass spectrometry, smFRET and/or EPR.
- in silico ligand docking and sequence optimization for stability.

The candidate must hold a PhD in the field of biology.

Contexte de travail

In-depth characterization of the NAD kinases is essential for understanding their role in cell biology. Bacterial NAD kinases have been shown to be essential and represent attractive therapeutic targets. More recent advances have suggested the role of human NAD kinases in cancer onset and metastasis.

In our group, we have defined the superfamily of enzymes to which NAD kinases belong. Since then we have solved numerous structure t suggests and then confirm an original substrate-based mechanism of catalysis shared by bacterial and human NAD kinases. We have also used an original fragment screening approach to design potent inhibitors.
More recently we have solved the first structure of the human cytoplasmic NAD kinase in its active form. We have demonstrated multiple levels of regulation by its N- and C-terminal extensions.
This post-doctoral project aims to bring more insights into the precise enzyme regulation of the NAD kinases and especially the human cytoplasmic one. These studies will rely on multiple techniques of structural biology such as X-ray crystallography, SAXS, cryo-electron microscopy, NMR and EPR. Time-resolved enzymology shall reveal important intermediate steps in the catalytic cycle. Allosteric effectors will be characterized as well as a potential regulation by membrane interactions. Conformational changes and flexibility will be explored by complementary techniques (SAXS, EPR, ensemble refinement, cryoEM, …).

Environment:
The CBS is a dynamic research institute with an outstanding infrastructure covering all techniques for structural biology and biophysics including NMR, X-ray crystallography, cryo-electron microscopy, advanced fluorescence and super-resolution microscopes, single-molecule manipulation, molecular and cell biology, and high-performance computation, allowing to carry out ambitious scientific projects by integrated and multi-scale approaches. Montpellier is a major biology area with a focus on basic research, biomedicine, plant biology and environmental science.

Through a position, funded by the Fondation pour la Recherche Médicale, the candidate will work in the “A.B.C.I.S.” group lead by Gilles Labesse and Corinne Lionne.
https://www.cbs.cnrs.fr/index.php/en/home-equipea4

Contraintes et risques

Interested candidates should upload their CV, a cover letter describing their research interests and motivation and the names of two people willing to write a recommendation letter.

Initial 12 month contract, renewable.

Informations complémentaires

Recent publications of the group related to this project:
Praharaj et al., Sci. Adv. 2025 doi: 10.1126/sciadv.ads2664