My Thesis

 

 

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Overview of my thesis

 

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0 -  Preparation of the starting materials

  • Design and manufacture of a versatile setup of pattern photobleaching using evanescent waves
  • Synthesis and purification of various transmembrane peptides ( with R.S. HODGES, CO, USA) (click here for details)
  • Design of a phase of bilayers for protein lateral diffusion and tests of cristallisation
  • Design and manufacture of microfluidics chips ( with T. THORSEN, MIT, Boston, USA)
  • labeling and reconstitution of transmembrane proteins in Giant Unilamellar Vesicles  ( list of the techniques here)

 

I -  Processes of diffusion :

 

II -  Lateral mobility in membranes      (click here for details)

  • Dependence in Radius of the protein, using transmembrane peptides and proteins : OmpA, OprM, and BR
  • Importance of the viscosity of the bounding fluid : effects of sugars, dextran, or PEGs on membranes.
  • Weight of the hydrophobic thickness of the membrane
  • Proposal of an heuristic law; possible theoritical model ( with N. GOV, Israel)

 

 III -  Understanding the anchorage of transmembrane peptides using single-molecule pulling experiments    (click here)

  • Role played by the hydrophobicity of the helix
  • Importance of the polar heads
  • Modelisation of the energy barriers explored by the peptides during their extraction
  • Deformation of membranes and threshold for microtubules formation;
  • dependence force-speed of extraction for the tubes

 

 IV -  Diverse consequences of the hydrophobic mismatch :       (to learn more...)

A) On the individual level :

  • anchorage of transmembrane peptides : the longer the stronger ?
  • Asymetric impact on lateral diffusion: "tilting" and "pinching" behaviors
  • Conformation of peptides as determined by IR Spectroscopy ( with F. HOMBLE, Belgium)
  • Determination of the critical mismatch for peptide insertion

B) Collective behavior controlled by mismatch :

  • auto-association of peptides in micro-patches for a critical mismatch
  • giant patches (up to 100 Ám) and spontaneous formation of vesicles

 

 V -  characterization of protein-protein interactions

  • Clustering of Bacteriorhodopsin upon photoactivation ( with P. BASSEREAU, Institut Curie, Paris, France)
  • Interaction between the Snares proteins VAMP- SNAP25 - Syntaxin (D. TARESTE, J. ROTHMAN, NY, USA)
  • Comparaison of the diffusion of CX3CR1 and CCR5 proteins in cells ( with P. DETERRE, Pitie-Salpetriere Hospital, Paris, France.)

 

 VI - Cristallisation of transmembrane proteins

  • Tests of isotropic, cubic-like phases of bilayers
  • Tests of progressive variation using microfluidics chips

 In collaboration with A. DUCRUIX, laboratoire de Cristallographie, Universite de Pharmacie, Paris.

 

If you want to know more, please let me know and I'd be happy to send you preprints or drafts...

 

 

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gambin@lps.ens.fr