Abstract：
:Even though a number of different in vitro fusion assays have been developed to analyze protein mediated fusion, they still only partially capture the essential features of the in vivo situation. Here we established an in vitro fusion assay that mimics the fluidity and planar geometry of the cellular plasma membrane to be able to monitor fusion of single protein-containing vesicles. As a proof of concept, planar pore-spanning membranes harboring SNARE-proteins were generated on highly ordered functionalized 1.2 μm-sized pore arrays in Si3N4. Full mobility of the membrane components was demonstrated by fluorescence correlation spectroscopy. Fusion was analyzed by two color confocal laser scanning fluorescence microscopy in a time resolved manner allowing to readily distinguish between vesicle docking, intermediate states such as hemifusion and full fusion. The importance of the membrane geometry on the fusion process was highlighted by comparing SNARE-mediated fusion with that of a minimal SNARE fusion mimetic.
journal_name
Sci Repjournal_title
Scientific reportsauthors
Schwenen LL,Hubrich R,Milovanovic D,Geil B,Yang J,Kros A,Jahn R,Steinem Cdoi
10.1038/srep12006subject
Has Abstractpub_date
2015-07-13 00:00:00pages
12006issn
2045-2322pii
srep12006journal_volume
5pub_type
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