post-doctoral researcher (M/F)

Job Description

Missions

The postdoctoral researcher will participate in the collaborative ANR-funded “FLIPPER” project, one of the objectives of which is to reconstitute in vitro the machinery responsible for the active transport of phospholipids. In cells, this transport is essential for maintaining the asymmetric distribution of phospholipids between the two leaflets of biological membranes. The main proteins involved in active lipid transport are lipid flippases, including the Drs2-Cdc50 complex.
It has been shown that the small GTPase Arl1 interacts with the N-terminal region of Drs2, while the Gea2 protein interacts with the C-terminal region of Drs2, and that both proteins stimulate lipid transport in vivo. The successful candidate will investigate in vitro, following reconstitution of the Drs2-Cdc50 complex into a model lipid system, the ability of Arl1 and Gea2 to activate lipid transport by Drs2-Cdc50. The candidate will also contribute to the production of a complex containing Gea2, Arl1, and Drs2-Cdc50 for subsequent structural characterization by cryo-electron microscopy.
The FLIPPER project brings together teams from the Institut de Génomique Fonctionnelle (IGF), Montpellier, Aarhus University (Denmark), and the Institut de Biologie Intégrative de la Cellule (I2BC), Gif-sur-Yvette.

Activity

Main activities
• Overexpression in S. cerevisiae and purification of the Drs2-Cdc50 complex and Gea2 protein using affinity chromatography and size-exclusion chromatography.
• Overexpression in E. coli and purification of the Arl1 protein using affinity chromatography and size-exclusion chromatography.
• Functional assays of purified proteins: measurement of ATP hydrolysis by Drs2-Cdc50 using a spectrophotometric assay, and measurement of nucleotide exchange activity by Gea2 using an assay based on the intrinsic fluorescence of Gea2.
• Reconstitution of the Drs2-Cdc50 complex into proteoliposomes.
• Measurement of Drs2-Cdc50-mediated lipid transport using fluorescently labeled lipids.
• Determination of the effects of Gea2 and Arl1 on phospholipid transport catalyzed by Drs2-Cdc50.
• Reconstitution of the Drs2-Cdc50-Gea2-Arl1 complex into nanodiscs for subsequent structural characterization by cryo-electron microscopy.
• Regular progress meetings with members of the research team and department, as well as with members of the FLIPPER consortium.

Your Profil

Skills

The successful candidate will hold a PhD in biochemistry or biophysics.
Essential technical skills
• Biochemistry and molecular biology, including cloning and site-directed mutagenesis.
Desirable technical skills
• Expression and purification of membrane proteins.
• Use of detergents for membrane protein solubilization.
• Reconstitution of membrane proteins into proteoliposomes.

The successful candidate should have excellent collaborative skills, be motivated and ambitious, and demonstrate the ability to take initiative and assume responsibilities within their working environment.

Your Work Environment

The successful candidate will join the Membrane Proteins and Systems (LPSM) team at I2BC. The LPSM investigates the molecular mechanisms underlying the transport of molecules and ions across biological membranes, using biochemical, biophysical, and molecular dynamics approaches. More specifically, the team studies the protein machinery responsible for maintaining the asymmetric distribution of lipids in biological membranes.
The LPSM is part of the Institut de Biologie Intégrative de la Cellule (I2BC), an interdisciplinary research institute comprising five scientific departments, approximately 60 research teams, and 15 high-level technological platforms. I2BC is located in Gif-sur-Yvette, approximately 25 km southwest of Paris, at the heart of Université Paris-Saclay.
The position is funded through a grant awarded by the French National Research Agency (ANR).

Constraints and risks

Some experimental protocols may require working outside regular working hours (7:00 a.m.–7:00 p.m.).

Compensation and benefits

Compensation

€3,132–€4,343 gross per month, depending on experience

Annual leave and RTT

44 jours

Remote Working practice and compensation

Pratique et indemnisation du TT

Transport

Prise en charge à 75% du coût et forfait mobilité durable jusqu’à 300€

About the offer

About the CNRS

The CNRS is a major player in fundamental research on a global scale. The CNRS is the only French organization active in all scientific fields. Its unique position as a multi-specialist allows it to bring together different disciplines to address the most important challenges of the contemporary world, in connection with the actors of change.

CNRS

The research professions