Date : 02/12/2011

Laboratory
Laboratoire de Biologie Physico-Chimique des Protéines Membranaires
UMR 7099 CNRS
IBPC
13 rue Pierre et Marie Curie
75005 Paris
Website
Main discipline : Biophysics
Lab director : Bruno MIROUX

PhD Supervisor
Dror WARSCHAWSKI
email : This e-mail address is being protected from spam bots, you need JavaScript enabled to view it
phone: +33 1 58 41 51 11

Subjects
1.: microbiology
2.: membrane biogenesis
3.: structural biology

Tools and Methodologies
1.: Nuclear Magnetic Resonance
2.: membrane biochemistry
3.: membrane protein expression

Summary of lab's interests

Our laboratory gathers biologists, physicists and chemists who are interested in the genesis, structure, structural dynamics and physical chemistry of membrane proteins, either in vivo within a lipid bilayer, or in vitro after solubilization. The biochemistry has extensive knowledge in the challenging overproduction of a wide range of membrane proteins in an active form, such as the cytochrome b6f complex, ATP synthase, mitochondrial uncoupling proteins, mechanosensitive channels, G protein-coupled receptors or porins... Among the structural determination techniques, X-ray crystallography, solution-state and solid-state NMR are being developed.

Summary of project

In recent years, our team has lead the first structural study of a membrane protein by solid-state NMR in France (Abdine, J Magn Reson 2010). We have produced this protein in E. coli and by in vitro synthesis (Abdine, New Biotechnol 2011). The expertise we have gained leads us to develop a new and ambitious approach: in cell solid-state NMR (Fu, J Am Chem Soc 2011), using strains developed and patented in the laboratory by Bruno Miroux (Miroux, J Mol Biol 1996), which overexpress the inner membrane and the subunit b of the E. coli ATP synthase. Several strains should be tested under conditions that allow for isotopic labeling of the proteins. Solid-state NMR will be tested on whole cells or purified native membranes of these bacteria. The goal is to optimize the sample preparation, to obtain the best spectral resolution possible, which can provide structural or dynamic information on the observed membrane proteins. This approach will be improved with the advent of a new D ynamic Nuclear Polarization spectrometer at 800 MHz, that is to be installed at ENS and that allows the detection of very small amount of biomolecules. This approach can then be transposed to other proteins that we study in the laboratory, such as mechanosensitive channels or GPCRs (G protein-coupled receptors, Catoire, J Am Chem Soc 2010).

Interdisciplinarity of the project

This project is both an advanced microbiology project (genetic improvement of bacterial strains, membrane protein structures...), and a solid-state NMR project, a technique that is currently growing in popularity. Those two skills are not easy to find in a student that is usually either trained in biochemistry or in physical-chemistry. The interdisciplinary partnership between our laboratory, the teams of Jean-Michel Betton (Institut Pasteur) and Geoffrey Bodenhausen (Ecole Normale Supérieure) will provide the ideal environment for such a project and for the student to develop both types of skills, including the first applications of high-field DNP in biological solids.

Do you have an available funding for this project ?

This project is eligible for an ANR grant that will be submitted early 2012, together with Jean-Michel Betton (Institut Pasteur).