Semisynthetic fluorescent sensor proteins


Project description
This project aims at the development of fluorescent sensor proteins for key intracellular metabolites and lipids. Up to now, genetically encoded fluorescent sensor proteins have been mostly used to probe the concentration of key metabolites in living cells. However, the approaches currently used to generate such fluorescent sensor proteins lack generality, as they require a protein that undergoes a conformational change upon metabolite binding. We have recently developed an approach that overcomes this limitation. Our biosensors consist of SNAP-tag, a fluorescent protein and a metabolite-binding protein. SNAP-tag is specifically labeled with a synthetic molecule containing a ligand of the metabolite-binding protein and a fluorophore. In the labeled sensor, the metabolite of interest displaces the intramolecular ligand from the binding protein, thereby shifting the sensor protein from a closed to an open conformation. The readout is a concomitant ratiometric change in the fluorescence intensities of the fluorescent protein and the tethered fluorophore. The observed ratiometric changes compare favorably with those achieved in genetically encoded fluorescent sensor proteins. In this project, we would like to apply this concept for the development of key intracellular metabolites, in particular sphingolipids.

Further reading
Brun et al., 2011, J Am Chem Soc 133(40):16235-42
Brun et al., 2009, J Am Chem Soc 131(16):5873-84

Host institute
Institute of Chemical Sciences and Engineering
École Polytechnique Fédérale de Lausanne
CH-1015 Lausanne
Phone: +41 (0)21 6939356

The host institute is one of the top-ranked European Technical Universities, located at the shore of beautiful Lake Geneva. It possess an excellent infrastructure and a highly international faculty and student population.

•    We are looking for an ambitious and interactive individual with MSc degree in biochemistry, chemistry or chemical biology. An interest in both chemistry as well biology is essential.
•    The candidate will be part of a multidisciplinary team of chemists and biochemists. Good communication skills are essential.
•    Proactive participation in SPHINGONET’s research & training program is expected through presentations and critical thinking at internal scientific forums, requiring excellent English presentation and writing skills. The ability to work in a team is a must.
•    Candidates must fulfill the eligibility criteria for Marie Curie Initial Training Networks (FP7).
•    Candidates should submit the complete application package.
•    Suitable candidates will be invited for an interview by the supervisor of the project.

•    PhD student for 3 years. Extension of the contract to 4 years is possible.
•    Full time (42 hrs/week).