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Reference : UMR5075-JERGAU-004
Workplace : GRENOBLE
Date of publication : Thursday, January 21, 2021
Type of Contract : FTC Scientist
Contract Period : 24 months
Expected date of employment : 1 May 2021
Proportion of work : Full time
Remuneration : According to experience and from € 2,648 gross
Desired level of education : PhD
Experience required : 1 to 4 years
The recruited candidate will investigate the structural dynamics of fluorescent proteins (FPs) used in super resolution fluorescence microscopy, using NMR, kinetic X-ray crystallography and single-molecule imaging.
The candidate will be recruited in the context of a recently awarded grant by the French “ANR”. FPs are the markers of choice for many applications in advanced fluorescence microscopy. Yet, super-resolution techniques such as PhotoActivation Localization Microscopy (PALM) and single-particle-tracking PALM (sptPALM) have not yet reached maturity, notably due to the non-ideal photophysical performance of the used FP labels. The project aims at enhancing the photostability of green-to-red photoconvertible fluorescent proteins (PCFPs) by combining investigations of their structural dynamics in realistic environmental conditions with large-scale engineering and screening at the single-molecule (SM) level.
The recruited candidate will concentrate on the mechanistic, structural biology part of the project. He/she will work as a bridge between two teams at the IBS (D.Bourgeois, Pixel team ; B. Brutscher, Biomolecular NMR) team) to perform :
- Molecular cloning, expression, purification and photophysical characterization of the FPs of interest.
- The preparation and optimization of crystals and their analysis by (kinetic) X-ray crystallography.
- The preparation and optimization of NMR sample and their analysis by a state-of-the-art NMR toolbox that has been recently developed at the IBS, specifically adapted to the study of fluorescent proteins.
- The interpretation of the X-ray and NMR data in terms of structural and dynamic behavior of the FPs.
- The preparation and optimization of FP samples and their analysis by absorption and fluorescence optical spectroscopy and by in vitro single molecule imaging.
- The suggestion of relevant mutation sites to engineer enhanced PCFPs (FP engineering itself will be performed by our project partners in Bordeaux (J.B. Sibarita, M. Sainlos)).
Overall, the candidate will seek to provide key mechanistic insights into the “dark side” of PCFPs, which has remained largely overlooked thus far due to its intricate nature, pushing the knowledge of FPs beyond the state-of-the-art.
Technical and scientific skills :
- Strong background in structural biophysics
- Strong background in multidimensional NMR spectroscopy, X-ray protein crystallography
- Good knowledge of biochemistry and molecular biology
- Good knowledge of fluorescence microscopy
- Additional expertise in cell biology, super-resolution microscopy, single-molecule imaging and fluorescent proteins will be key advantages.
- Knowing how to work in a team
- Master the techniques of oral and written presentations, write reports of the experiments carried out, play a leading role in the writing of scientific articles
- Have the ability to work in an international and multidisciplinary environment
- Have initiative and organizational skills, be rigorous, participate in the collective tasks of the laboratory
- Minimum level of English B2 in comprehension and speaking
IBS is a research unit affiliated to the University Grenoble-Alpes, CNRS and CEA. It currently employs 280 people, including many students and post-doctoral fellows from different countries. It has high-performance and internationally competitive facilities in molecular and cell biology, biochemistry, biophysics and structural biology. The Institute is a founding member of the Structural Biology Partnership with prestigious pan-European institutes: the European Synchrotron Radiation Source (ESRF), the European Neutron Scattering Center (ILL), and the EMBL Grenoble station, all located nearby and offering a strong international environment.
Within IBS, the recruited person will join Dominique Bourgeois's « Pixel » team (https://www.ibs.fr/research/research-groups/integrated-imaging-of-stress-response-group/pixel-559/)
The successful candidate will be recruited for an initial period of 2 years, with a possibility of extension for 1 additional year.
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