Marie Skłodowska-Curie Action
Innovative Training Networks (ITN)

2015-2019

 

Funded by

marie curie actions   Marie Curie Actions

 

 

unnamed   
EU Horizon 2020

 

 

european union logo   European Union

 

Functional proteomics of CXCR4 and CXCR7-associated signalling networks

Early stage researcher 12 (ESR12) project
Supervision: Dr P Marin, Dr M Séveno, Dr S Chaumont-Dubel, Dr S Urbach
Host: CNRS (FR) – Institut de Génomique Fonctionnelle, CNRS UMR 5203, INSERM U1191,

Université de Montpellier



I- Project proposal:

Aim:

1. Identify intracellular partners (GIPs) of CXCR4 and CXCR7 using an AP-MS proteomics strategy.

2. Determine functional consequences of CXCR4 and CXCR7 association with identified GIPs.

3. Decipher the phosphoproteomes resulting from CXCR4 and CXCR7 activation by different modulators.



Methodology:
Proteins interacting with epitope-tagged versions of CXCR4 and CXCR7 receptors will be purified by immunoprecipitation and affinity-purified proteins will be systematically identified by Nano-flow liquid chromatography coupled with Fourier transform tandem mass spectrometry (Nano-LC-FT-MS/MS). The influence of identified partners on receptor-operated signal transduction will be investigated by overexpressing or silencing expression of candidate partners and using tools and/or biosensors developed in this consortium. Phosphoproteomes generated upon receptor activation by different modulators will be deciphered by combining Stable Isotope Labelling by Amino acids in Cultured cells (SILAC) and a double-phosphopeptide enrichment procedure consisting of Hydrophilic Interaction Liquid Chromatography (HILIC) followed by Immobilized Metal Affinity Chromatography (IMAC). Planned secondments: Actelion (CH), Vivia Biosystems (ES)



II- Requirement candidate:
Required diploma: MSc degree in molecular/biomedical Life Sciences, Pharmaceutical Sciences or related Life Science degree.

Required expertise: cell culture, cell-based assays, biochemistry, molecular biology. Recommended expertise: Basic knowledge in mass spectrometry, proteomics, protein databases, bioinformatics and statistics.



Key publications:
1. Duhr, F., Deleris, P., Raynaud, F., Seveno, M., Morisset-Lopez, S., Mannoury la Cour, C., Millan, M. J., Bockaert, J., Marin, P. & Chaumont-Dubel, S. (2014) Cdk5 induces constitutive activation of 5-HT6 receptors to promote neurite growth Nat Chem Biol 10, 590-597. News and Views: Seo and Tsai. Neuronal differentiation: 5-HT6R can do it alone. Nat Chem Biol 10, 590-597.

2. Karaki, S., Becamel, C., Murat, S., Mannoury la Cour, C., Millan, M. J., Prezeau, L., Bockaert, J., Marin, P. & Vandermoere, F. (2014) Quantitative phosphoproteomics unravels biased phosphorylation of serotonin 2A receptor at Ser280 by hallucinogenic versus nonhallucinogenic agonists. Mol Cell Proteomics 13, 1273-1285.

3. Meffre, J., Chaumont-Dubel, S., Mannoury la Cour, C., Loiseau, F., Watson, D. J., Dekeyne, A., Seveno, M., Rivet, J. M., Gaven, F., Deleris, P., Herve, D., Fone, K. C., Bockaert, J., Millan, M. J. & Marin, P. (2012) 5-HT(6) receptor recruitment of mTOR as a mechanism for perturbed cognition in schizophrenia. EMBO Mol Med 4, 1043-1056.

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