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

Dana Bar-On, Ulrike Winter, Esther Nachliel, Menachem Gutman, Dirk Fasshauer, Thorsten Lang, Uri Ashery

Research output: Contribution to journalArticlepeer-review

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 (α-SNAP) concentrations.

Original languageEnglish
Pages (from-to)3563-3568
Number of pages6
JournalFEBS Letters
Volume582
Issue number23-24
DOIs
StatePublished - 15 Oct 2008

Keywords

  • Assembly
  • Disassembly
  • Ex vivo
  • Plasma membrane sheet
  • Time-resolved kinetics
  • cis-SNARE complex

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