Post-doc position: New biosensors for spatio-temporal detection of phagosomal ROS production
Neutrophils are professional phagocytes of our innate immune system that trap pathogens in intracellular vesicles called phagosomes where they can deliver all components necessary for their killing. In particular, the phagocyte is able to activate two enzymatic systems NADPH oxidase (NOX2) and myeloperoxidase (MPO) that produce in the phagosome large amounts of reactive oxygen species (ROS) (scheme). [1] Little is known about their quantity and the timing of their production mainly because of the lack of analytical tools compatible with the observation of a closed and highly dynamic compartment like the phagosome. [1] Nevertheless we already proved that fluorescence microscopy techniques offer the possibility to follow individual phagosomes, access to spatial and temporal details and eventually offer quantitative analysis. [2] To go further this post doc fellowship aims to develop new genetically encoded ROS biosensors based on cyan variants (CFPs) of the Green Fluorescent Protein (GFPav). Taking advantage of our in depth knowledge of the cyan variants photophysic and reactivity, CFPs will be rationally engineered toward a biosensor with the appropriate properties. An identical workflow recently allowed us to produce a new bright CFP with improved performances. [3] This new biosensor that could be directly expressed by the pathogen itself will also be compatible with the monitoring of NOX2 assembly by microscopy techniques. The combined observation of ROS and NOX2 assembly will finally give new insights on the role of neutrophils in the development of infectious diseases.
The candidate will have a strong background in molecular biology and biochemistry as well as skills in fluorescence spectroscopy and microscopy. The candidate will also have basic knowledge in cell biology. Some expertise on the structure-function relationship of Fluorescent Protein of the GFPav family will be appreciated. The candidate will also show interest for working in a multidisciplinary field with two complementary teams on the same campus.
The Post-doc position is funded by the “Fondation pour la Recherche Médicale” for 2 year (http://www.frm.org/) starting from November 1st, 2013.
Applications containing a detailed CV including an abstract of the PhD thesis, a description of research experience and interests and names of two references should be send by email to:
Marie Erard (LCP, biosensor and fluorescent protein): marie.erard@u-psud.fr
Oliver Nüsse (UMR S1174, phagocytes and cell biology): oliver.nusse@u-psud.fr
Web site: https://www.scicf.universite-paris-saclay.fr and http://www.lcp.u-psud.fr
[1] Nüsse O Thescientificworldjournal 2011 (11) 2364
[2] Tlili A Free Rad Bio Med 2011 (50) 438. Tlili A J Leuk Biol 2012 (91) 83
[3] Fredj A Plos One 2012 (7) e49149, Erard M Mol BioSyst 2013 (9) 258