Sequential N- to C-terminal SNARE complex assembly drives priming and fusion of secretory vesicles.

Détails

ID Serval
serval:BIB_328AE15F979C
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Sequential N- to C-terminal SNARE complex assembly drives priming and fusion of secretory vesicles.
Périodique
EMBO Journal
Auteur(s)
Sørensen J.B., Wiederhold K., Müller E.M., Milosevic I., Nagy G., de Groot B.L., Grubmüller H., Fasshauer D.
ISSN
0261-4189 (Print)
ISSN-L
0261-4189
Statut éditorial
Publié
Date de publication
2006
Peer-reviewed
Oui
Volume
25
Numéro
5
Pages
955-966
Langue
anglais
Résumé
During exocytosis a four-helical coiled coil is formed between the three SNARE proteins syntaxin, synaptobrevin and SNAP-25, bridging vesicle and plasma membrane. We have investigated the assembly pathway of this complex by interfering with the stability of the hydrophobic interaction layers holding the complex together. Mutations in the C-terminal end affected fusion triggering in vivo and led to two-step unfolding of the SNARE complex in vitro, indicating that the C-terminal end can assemble/disassemble independently. Free energy perturbation calculations showed that assembly of the C-terminal end could liberate substantial amounts of energy that may drive fusion. In contrast, similar N-terminal mutations were without effects on exocytosis, and mutations in the middle of the complex selectively interfered with upstream maturation steps (vesicle priming), but not with fusion triggering. We conclude that the SNARE complex forms in the N- to C-terminal direction, and that a partly assembled intermediate corresponds to the primed vesicle state.
Mots-clé
Amino Acid Sequence, Amino Acid Substitution, Animals, Binding Sites, Cell Membrane/metabolism, Chromaffin Cells/cytology, Chromaffin Cells/metabolism, Circular Dichroism, Electrophysiology, Exocytosis, Membrane Fusion, Mice, Mice, Knockout, Molecular Sequence Data, Mutation, Qa-SNARE Proteins/metabolism, R-SNARE Proteins/metabolism, Secretory Vesicles/chemistry, Secretory Vesicles/metabolism, Sequence Homology, Amino Acid, Synaptosomal-Associated Protein 25/metabolism
Pubmed
Web of science
Open Access
Oui
Création de la notice
15/09/2011 9:21
Dernière modification de la notice
20/08/2019 14:18
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