title: 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.
authors.affiliation: Department of Neurobiochemistry, Tel Aviv University, Tel Aviv 69978, Israel.
authors.forename: Dana
authors.initials: D
authors.lastname: Bar-On
authors.forename: Ulrike
authors.initials: U
authors.lastname: Winter
authors.forename: Esther
authors.initials: E
authors.lastname: Nachliel
authors.forename: Menachem
authors.initials: M
authors.lastname: Gutman
authors.forename: Dirk
authors.initials: D
authors.lastname: Fasshauer
authors.forename: Thorsten
authors.initials: T
authors.lastname: Lang
authors.forename: Uri
authors.initials: U
authors.lastname: Ashery
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000737:chemistry* / Q000648:ultrastructure; D007700:Kinetics; D008856:Microscopy, Fluorescence / Q000379:methods*; D052066:N-Ethylmaleimide-Sensitive Proteins / Q000737:chemistry; D016716:PC12 Cells; D051381:Rats; D050600:SNARE Proteins / Q000737:chemistry*; D013997:Time Factors
chemical_list: D050600:SNARE Proteins; D052066:N-Ethylmaleimide-Sensitive Proteins; C498737:Nsf protein, rat