Berlin 2015 – scientific programme
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BP: Fachverband Biologische Physik
BP 18: Membranes and vesicles I (joint BP/CPP)
BP 18.3: Talk
Tuesday, March 17, 2015, 10:15–10:30, H 1028
New Strategy to Study a Single SNARE Mediated Membrane Fusion Event — •Jose Nabor Vargas1, Kewin Howan2, Andrea Gohlke2,4, Ralf Seemann1,3, Jean-Baptiste Fleury1, and Frederic Pincet2,4 — 1Experimental Physics, Saarland University, Saarbrücken, Germany — 2Laboratoire de Physique Statistique, Ecole Normale Supérieure, 75005 Paris, France — 3Max Planck Institute for Dynamics and Self-Organization, Göttingen, Germany — 4Department of Cell Biology, School of Medicine, Yale University, CT 06520 New Haven, USA
We present an approach to explore the properties of a single SNARE mediated membrane fusion event in a microfluidic chip. In a first step, a single free standing lipid membrane is generated at a defined position with the Droplet Interface Bilayer technique (DiB). In a second step, we inject a solution of divalent cations (Calcium, Ca2+) and small unilamellar vesicles functionalized with T-SNARE proteins (T-SUVs) around the planar membrane using a volume controlled flow. The presence of calcium mediates the direct fusion of the vesicles with the planar membrane, which is incorporating the proteins into the membrane. In a third step, we remove the calcium and the T-SUVs with a buffer solution. After this washing step, a solution of small unilamellar vesicles functionalized with V-SNARE proteins (V-SUVs) is injected around the planar membrane. And finally, we study single fusion event with good optical and electrical access.