Imaging the assembly and disassembly kinetics of cis-SNARE complexes on native plasma membranes.

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.

journal_name

FEBS Lett

journal_title

FEBS letters

authors

Bar-On D,Winter U,Nachliel E,Gutman M,Fasshauer D,Lang T,Ashery U

doi

10.1016/j.febslet.2008.08.040

subject

Has Abstract

pub_date

2008-10-15 00:00:00

pages

3563-8

issue

23-24

eissn

0014-5793

issn

1873-3468

pii

S0014-5793(08)00771-0

journal_volume

582

pub_type

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