English
 
User Manual Privacy Policy Disclaimer Contact us
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT

Released

Journal Article

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

MPS-Authors
/persons/resource/persons104871

Lindau,  M.
Research Group of Nanoscale Cell Biology, MPI for Biophysical Chemistry, Max Planck Society;

External Ressource
No external resources are shared
Fulltext (public)

2418532.pdf
(Publisher version), 3MB

Supplementary Material (public)

2418532_Suppl.zip
(Supplementary material), 2MB

Citation

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.


Cite as: http://hdl.handle.net/11858/00-001M-0000-002C-EA3C-7
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.