Single vesicle millisecond fusion kinetics reveals number of SNARE complexes optimal for fast SNARE-mediated membrane fusion.

Détails

Ressource 1Télécharger: Domanskaetal2009.pdf (2813.02 [Ko])
Etat: Public
Version: Final published version
ID Serval
serval:BIB_BF5B7B4971D2
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Single vesicle millisecond fusion kinetics reveals number of SNARE complexes optimal for fast SNARE-mediated membrane fusion.
Périodique
Journal of Biological Chemistry
Auteur(s)
Domanska M.K., Kiessling V., Stein A., Fasshauer D., Tamm L.K.
ISSN
1083-351X (Electronic)
ISSN-L
0021-9258
Statut éditorial
Publié
Date de publication
2009
Peer-reviewed
Oui
Volume
284
Numéro
46
Pages
32158-32166
Langue
anglais
Résumé
SNAREs mediate membrane fusion in intracellular vesicle traffic and neuronal exocytosis. Reconstitution of membrane fusion in vitro proved that SNAREs constitute the minimal fusion machinery. However, the slow fusion rates observed in these systems are incompatible with those required in neurotransmission. Here we present a single vesicle fusion assay that records individual SNARE-mediated fusion events with millisecond time resolution. Docking and fusion of reconstituted synaptobrevin vesicles to target SNARE complex-containing planar membranes are distinguished by total internal reflection fluorescence microscopy as separate events. Docking and fusion are SNAP-25-dependent, require no Ca(2+), and are efficient at room temperature. Analysis of the stochastic data with sequential and parallel multi-particle activation models reveals six to nine fast-activating steps. Of all the tested models, the kinetic model consisting of eight parallel reaction rates statistically fits the data best. This might be interpreted by fusion sites consisting of eight SNARE complexes that each activate in a single rate-limiting step in 8 ms.
Mots-clé
Animals, Exocytosis, Kinetics, Lipid Bilayers/metabolism, Membrane Fluidity, Membrane Fusion, Microscopy, Fluorescence, Microscopy, Interference, Models, Molecular, Protein Binding, Proteolipids/chemistry, Proteolipids/metabolism, R-SNARE Proteins/metabolism, Rats, SNARE Proteins/metabolism, Synaptic Vesicles/metabolism, Synaptosomal-Associated Protein 25/metabolism
Pubmed
Web of science
Création de la notice
15/09/2011 8:52
Dernière modification de la notice
20/08/2019 16:33
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