Group leader : Jacqueline Cherfils
Our aim is to understand how molecular reactions govern the function of small GTPases of the Arf, Rab and Rho families in cellular traffic and cytoskeleton dynamic in normal cells and within pathological contexts. These proteins are ubiquitous molecular switches in eukaryotes and their regulation mechanisms are emerging as therapeutic targets, particularly in cancers. By using a combination of structural and cellular biochemistry methods, we observe these proteins at the atomic level, we analyze their interactions with other proteins and with membranes, we characterize their specificity and we decipher their regulation mechanisms. Our recent results describe the activation of small GTPases of the Arf family by exchange factors involved in cancers and infections, and their inhibition by small chemical inhibitors with therapeutic potential.
Free download of our article "Regulation of small GTPases by GEFs, GAPs and GDIs", Jacqueline Cherfils and Mahel Zeghouf, Physiological Reviews 2013
- Production and purification of recombinant proteins: cloning, directed mutagenesis, production (E. coli and insect cells) and chromatography.
- Biophysics of proteins: circular dichroism, dynamic light scattering, SEC-MALS
- Structure of protein complexes: crystallization, crystallography, small angle X-ray scattering (SAXS), structure analysis
- Protein-protein interactions: chromatography, fluorescence anisotropy, SEC-MALS, microscale thermophoresis, calorimetry
- Protein-membrane interactions: liposomes manipulation, fluorescence spectroscopy
- Reaction mechanisms: fluorescence kinetics
- Inhibitor characterization: fluorescence spectroscopy, microplate format analysis
- Cellular biology: mammalian cell culture, immunofluorescence, confocal microscopy
Campanacci V, Mukherjee S, Roy CR, Cherfils J. Structure of the Legionella effector AnkX reveals the mechanism of phosphocholine transfer by the FIC domain. EMBO J (2013) accepted
Cherfils J, Zeghouf M. Regulation of Small GTPases by GEFs, GAPs, and GDIs. Physiol Rev. (2013) 93:269-309.
Alix E., Chesnel L., Bowzard B.J., Tucker A.M., Delprato A., Cherfils J., Wood D.O, Kahn R.A. and Roy C.R. The capping domain in RalF regulates effector functions. PLoS Pathog. (2012) Nov;8(11):e1003012.
Vigil, D, Cherfils J, Rossman KL and Der CJ. Ras family GEFs and GAPs: validated and tractable targets for cancer therapy. Nature Reviews Cancer 2010, 10, 842-857.
Biou V, Aizel K, Roblin P, Thureau A, Jacquet E, Hansson S, Guibert B, Guittet E, van Heijenoort C, Zeghouf M, Perez J, Cherfils J. SAXS and X-ray Crystallography Suggest an Unfolding Model for the GDP/GTP Conformational Switch of the Small GTPase Arf6. J Mol Biol. 2010 Oct 1;402(4):696-707
Ramaen, O., A. Joubert, P. Simister, N. Belgareh-Touze, M.C. Olivares-Sanchez, J.C. Zeeh, S. Chantalat, M.P. Golinelli-Cohen, C.L. Jackson, V. Biou, and J. Cherfils, Interactions between conserved domains within homodimers in the BIG1, BIG2, and GBF1 Arf guanine nucleotide exchange factors. J Biol Chem, 2007. 282(39): p. 28834-42.
Pommier, Y. and J. Cherfils, Interfacial inhibition of macromolecular interactions: nature's paradigm for drug discovery. Trends Pharmacol Sci, 2005. 26(3): p. 138-45.
Pasqualato, S., F. Senic-Matuglia, L. Renault, B. Goud, J. Salamero, and J. Cherfils, The structural GDP/GTP cycle of Rab11 reveals a novel interface involved in the dynamics of recycling endosomes. J Biol Chem, 2004. 279(12): p. 11480-8.
Renault, L., B. Guibert, and J. Cherfils, Structural snapshots of the mechanism and inhibition of a guanine nucleotide exchange factor. Nature, 2003. 426(6966): p. 525-30. Cité dans faculty of 1000.
Ménétrey, J., E. Macia, S. Pasqualato, M. Franco, and J. Cherfils, Structure of Arf6-GDP suggests a basis for guanine nucleotide exchange factors specificity. Nat Struct Biol, 2000. 7(6): p. 466-9.
Cherfils, J., J. Ménétrey, M. Mathieu, G. Le Bras, S. Robineau, S. Béraud-Dufour, B. Antonny, and P. Chardin, Structure of the Sec7 domain of the Arf exchange factor ARNO. Nature, 1998. 392(6671): p. 101-5.