Imaging the assembly and disassembly kinetics of cis-SNARE complexes on native plasma membranes.
Details
Serval ID
serval:BIB_DF53B018DD0C
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Imaging the assembly and disassembly kinetics of cis-SNARE complexes on native plasma membranes.
Journal
FEBS Letters
ISSN
0014-5793 (Print)
ISSN-L
0014-5793
Publication state
Published
Issued date
2008
Peer-reviewed
Oui
Volume
582
Number
23-24
Pages
3563-3568
Language
english
Abstract
Mild sonication of eukaryotic cells produces native plasma membrane sheets that retain their docked organelles, cytoskeleton structures and cytoplasmic complexes. While the delicate organization of membranous protein complexes remains undisturbed, their inner plasmalemmel leaflet can be rapidly exposed to bathing solutions, enabling specific biochemical manipulations. Here, we apply this system to track membrane-biochemistry kinetics. We monitor soluble NSF-attachment protein receptor (SNARE) complex assembly and disassembly on the plasma membrane at high time resolution. The results suggest two-phase kinetics for the assembly process and dependence of the disassembly kinetics on both N-ethyl maleimide-sensitive factor (NSF) and soluble NSF-attachment protein (alpha-SNAP) concentrations.
Keywords
Animals, Cell Membrane/chemistry, Cell Membrane/ultrastructure, Kinetics, Microscopy, Fluorescence/methods, N-Ethylmaleimide-Sensitive Proteins/chemistry, PC12 Cells, Rats, SNARE Proteins/chemistry, Time Factors
Pubmed
Web of science
Open Access
Yes
Create date
15/09/2011 8:06
Last modification date
20/08/2019 16:03