Des bicouches suspendues pour l’étude de la cinétique et de l’organisation protéique dans la fusion membranaire médiée par les SNARE. – SuspenSNARE

SNARE induced membrane fusion is involved at every level in intra-cellular traffic. However, molecular details of this fusion machinery such as kinetics and molecular organization remain largely unknown. We propose, thanks to a tight collaboration between our complementary teams, to use a novel approach that we are currently designing. In this approach, transmembrane SNARE proteins are inserted into a suspended bilayer which will allow an artifact free study of single membrane fusion events by evanescent field microscopy (TIRF). We already have preliminary results showing that our new approach works and will be usable by many other scientist teams. In this project, we will completely finish the setup and characterization of this technique and apply it SNARE induced fusion. Namely, we will focus on the kinetics of association of the proteins, their organization prior to fusion, and the kinetics and molecular role of regulatory proteins: Synaptotagmin, Complexin and Munc18.