Early stage researcher 3 (ESR3) project
Supervision: Dr. Thierry Durroux, Dr. Sébastien Granier
Host: CNRS 

I- Project proposal:

Aim:

1. Analysis of the pharmacological properties of homo/hetero-oligomers of CXCR4/ACKR3/CCR5

2. Impact of homo- or hetero-oligomerization on CXCR4 structural properties
   

 Methodology:

1. Pharmacological properties will be investigated using time-resolved FRET- or BRET-based strategies with fluorescent ligands or nanobodies. Binding experiments will be conducted using the Tag-Lite approach (TR-FRET based). Signalling properties will be examined, looking at G protein coupling using G protein BRET biosensors, arrestin association and arrestin-mediated signalling, internalisation and recycling TR-FRET assays.
2. Structural analysis of the complexes will be performed in collaboration with Gaetan Bellot, a biophysicists. Biochemical strategies (for receptor purification and electronic microscopy will be implemented. Electronic microscopy will be performed to investigate receptor structure

 
Planned secondments:

Cisbio Bioassays

II – Requirement candidate:

Required diploma: MSc in molecular/biochemical Life Sciences, pharmaceutical sciences or related Life-Science degree.

Required expertise: biochemistry, pharmacology and molecular biology. 

Recommended expertise: bases in molecular biology, biochemistry, cell culture, cell based assays, fluorescence microscopy.

Key publications:  

1. Vasiliauskaité-Brooks I, Healey RD, Rochaix P, Saint-Paul J, Sounier R, Grison C, Waltrich-Augusto T, Fortier M, Hoh F, Saied EM, Arenz C, Basu S, Leyrat C, Granier S. Structure of a human intramembrane ceramidase explains enzymatic dysfunction found in leukodystrophy. Nat Commun. 2018 Dec 21;9(1):5437.
2. Vasiliauskaité-Brooks I, Sounier R, Rochaix P, Bellot G, Fortier M, Hoh F, De Colibus L, Bechara C, Saied EM, Arenz C, Leyrat C, Granier S. Structural insights into adiponectin receptors suggest ceramidase activity. Nature. 2017 Apr 6;544(7648):120-123
3. Hounsou C, Baehr C, Gasparik V, Alili D, Belhocine A, Rodriguez T, Dupuis E, Roux T, Mann A, Heissler D, Pin JP, Durroux T, Bonnet D, Hibert M. From the Promiscuous Asenapine to Potent Fluorescent Ligands Acting at a Series of Aminergic G-Protein-Coupled Receptors. J Med Chem. 2018 61:174-188
4. Faklaris O, Cottet M, Falco A, Villier B, Laget M, Zwier JM, Trinquet E, Mouillac B, Pin JP, Durroux T. Multicolor time-resolved Förster resonance energy transfer microscopy reveals the impact of GPCR oligomerization on internalization processes. FASEB J. 2015 29(6):2235-46.
5. Albizu L, Cottet M, Kralikova M, Stoev S, Seyer R, Brabet I, Roux T, Bazin H, Bourrier E, Lamarque L, Breton C, Rives ML, Newman A, Javitch J, Trinquet E, Manning M, Pin JP, Mouillac B, Durroux T. Time-resolved FRET between GPCR ligands reveals oligomers in native tissues. Nat Chem Biol. 2010 6: 587-94.

For more informetion:
Dr. Thierry Durroux – thierry.durroux@igf.cnrs.fr

Dr. Sébastien Granier – sebastien.granier@igf.cnrs.fr  

Application:

Please send your application to sebastien.granier@igf.cnrs.fr and thierry.durroux@igf.cnrs.fr 

Include in heading – ONCORNET2.0 application + ESR #