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  Resolving single membrane fusion events on planar pore-spanning membranes.

Schwenen, L. L. G., Hubrich, R., Milovanovic, D., Geil, B., Yang, J., Kros, A., et al. (2015). Resolving single membrane fusion events on planar pore-spanning membranes. Scientific Reports, 5: 12006. doi:10.1038/srep12006.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0028-1956-5 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-002A-2288-E
Genre: Journal Article

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Schwenen, L. L. G., Author
Hubrich, R., Author
Milovanovic, D.1, Author              
Geil, B., Author
Yang, J., Author
Kros, A., Author
Jahn, R.1, Author              
Steinem, C., Author
Affiliations:
1Department of Neurobiology, MPI for biophysical chemistry, Max Planck Society, ou_578595              

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 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 mu m-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.

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Language(s): eng - English
 Dates: 2015-07-13
 Publication Status: Published online
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 Rev. Method: Peer
 Identifiers: DOI: 10.1038/srep12006
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Title: Scientific Reports
Source Genre: Journal
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Pages: 15 Volume / Issue: 5 Sequence Number: 12006 Start / End Page: - Identifier: -