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  SNARE-mediated membrane fusion trajectories derived from force-clamp experiments.

Oelkers, M., Witt, H., Halder, P., Jahn, R., & Janshoff, A. (2016). SNARE-mediated membrane fusion trajectories derived from force-clamp experiments. Proceedings of the National Academy of Sciences of the United States of America, 113(46), 13051-13056. doi:10.1073/pnas.1615885113.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-002C-3197-7 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-002C-7EA9-C
Genre: Journal Article

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 Creators:
Oelkers, M., Author
Witt, H., Author
Halder, P.1, Author              
Jahn, R.1, Author              
Janshoff, A., Author
Affiliations:
1Department of Neurobiology, MPI for Biophysical Chemistry, Max Planck Society, ou_578595              

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Free keywords: membrane fusion; SNARE; force spectroscopy; AFM; hydration barrier
 Abstract: Fusion of lipid bilayers is usually prevented by large energy barriers arising from removal of the hydration shell, formation of highly curved structures, and, eventually, fusion pore widening. Here, we measured the force-dependent lifetime of fusion intermediates using membrane-coated silica spheres attached to cantilevers of an atomic-force microscope. Analysis of time traces obtained from force-clamp experiments allowed us to unequivocally assign steps in deflection of the cantilever to membrane states during the SNARE-mediated fusion with solid-supported lipid bilayers. Force-dependent lifetime distributions of the various intermediate fusion states allowed us to propose the likelihood of different fusion pathways and to assess the main free energy barrier, which was found to be related to passing of the hydration barrier and splaying of lipids to eventually enter either the fully fused state or a long-lived hemifusion intermediate. The results were compared with SNARE mutants that arrest adjacent bilayers in the docked state and membranes in the absence of SNAREs but presence of PEG or calcium. Only with the WT SNARE construct was appreciable merging of both bilayers observed.

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Language(s): eng - English
 Dates: 2016-11-15
 Publication Status: Published online
 Pages: -
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 Table of Contents: -
 Rev. Method: Peer
 Identifiers: DOI: 10.1073/pnas.1615885113
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Title: Proceedings of the National Academy of Sciences of the United States of America
Source Genre: Journal
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Pages: - Volume / Issue: 113 (46) Sequence Number: - Start / End Page: 13051 - 13056 Identifier: -