PhD position in NMR spectroscopy of photoswitchable fluorescent proteins at the Institute of Structural Biology (IBS) Grenoble, France
COMBINING IN-SITU ILLUMINATION AND OXYGEN PRESSURE CONTROL WITH HIGH RESOLUTION NMR SPECTROSCOPY FOR UNDERSTANDING AND IMPROVING THE PROPERTIES OF PHOTOSWITCHABLE FLUORESCENT PROTEINS
The recruited student will investigate the photophysical mechanisms of reversibly photoswitchable fluorescent proteins employing solution NMR spectroscopy coupled with in- situ illumination and variable oxygen pressure. The goal will be to contribute fundamental knowledge of these fluorescent markers and to design improved variants.
Advanced fluorescence imaging is essential to discover the secrets of life, and has largely benefited from the discovery of Fluorescent Proteins (FPs) and other protein-based fluorescent markers. Reversibly Switchable Fluorescent Proteins (RSFPs, Ref 1) are capable to switch between a fluorescent on-state and a nonfluorescent off-state under specific light illumination, and have fostered many types of imaging applications including super-resolution methods. Yet, RSFPs are still imperfect: e.g. their brightness is limited, their switching kinetics is perturbed by the involvement of multiple photophysical states, their resistance to irreversible photobleaching is lower than that of typical organic dyes.
Multidimensional NMR spectroscopy is particularly powerful in providing detailed information on conformational protein dynamics and the local chemistry (protonation states, H-bonding interactions) involved in the photophysics of the chomophore (Ref 2 and 3). In the proposed PhD project, we intend to further improve our NMR in-situ illumination device by adding capabilities such as additional wavelengths of emitting light sources, fluorescence detection, and oxygen pressure control. This will allow to directly correlate the conformational dynamics of various states with their photophysical properties, as well as the effect of oxygen on triplet state formation and photobleaching. We will apply this NMR methodology to several green and red model RSFPs, as well as FAST systems (Ref 4). The recruited student will work in close collaboration with another PhD student, who will approach the same questions by employing optical spectroscopy, X-ray crystallography and fluorescence microscopy.
Grenoble is situated in the middle of the beautiful French Alps, and the IBS provides a unique environment for state-of-the-art integrated cellular and structural biology (http://www.ibs.fr/).
Candidates should have a strong interest to work at the interface between physics, chemistry and biology. Some basic knowledge of NMR spectroscopy will be appreciated.
The project is financed by the French ANR (project: Photoswitch NMR), and the student will be employed by CEA (https://instn.cea.fr/en/theses-and-phds-at-the-cea/). Gross salary is ~2400€ per month.
Applications are now open. Please send a CV, a detailed motivation letter, your transcripts, and at least one reference letter to Bernhard Brutscher (bernhard.brutscher@ibs.fr). Selection will stop when a suitable candidate has been identified.
