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  AFM/TIRF force clamp measurements of neurosecretory vesicle tethers reveal characteristic unfolding steps.

Harris, M. C., Cislo, D., Lenz, J. S., Umbach, C., & Lindau, M. (2017). AFM/TIRF force clamp measurements of neurosecretory vesicle tethers reveal characteristic unfolding steps. PLoS One, 12(3): e0173993. doi:10.1371/journal.pone.0173993.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-002C-EA3C-7 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-002C-EA40-C
Genre: Journal Article

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 Creators:
Harris, M. C., Author
Cislo, D., Author
Lenz, J. S., Author
Umbach, C., Author
Lindau, M.1, Author              
Affiliations:
1Research Group of Nanoscale Cell Biology, MPI for Biophysical Chemistry, Max Planck Society, ou_1832294              

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 Abstract: Although several proteins have been implicated in secretory vesicle tethering, the identity and mechanical properties of the components forming the physical vesicle-plasma membrane link remain unknown. Here we present the first experimental measurements of nanomechanical properties of secretory vesicle-plasma membrane tethers using combined AFM force clamp and TIRF microscopy on membrane sheets from PC12 cells expressing the vesicle marker ANF-eGFP. Application of pulling forces generated tether extensions composed of multiple steps with variable length. The frequency of short (<10 nm) tether extension events was markedly higher when a fluorescent vesicle was present at the cantilever tip and increased in the presence of GTPγS, indicating that these events reflect specifically the properties of vesicle-plasma membrane tethers. The magnitude of the short tether extension events is consistent with extension lengths expected from progressive unfolding of individual helices of the exocyst complex, supporting its direct role in forming the physical vesicle-plasma membrane link.

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Language(s): eng - English
 Dates: 2017-03-21
 Publication Status: Published online
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 Rev. Method: Peer
 Identifiers: DOI: 10.1371/journal.pone.0173993
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Title: PLoS One
Source Genre: Journal
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Pages: 20 Volume / Issue: 12 (3) Sequence Number: e0173993 Start / End Page: - Identifier: -