Interactions between synaptic vesicle fusion proteins explored by atomic force microscopy

Détails

ID Serval
serval:BIB_0ACAD6545897
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
Interactions between synaptic vesicle fusion proteins explored by atomic force microscopy
Périodique
Proceedings of the National Academy of Sciences of the United States of America
Auteur(s)
Yersin  A., Hirling  H., Steiner  P., Magnin  S., Regazzi  R., Huni  B., Huguenot  P., De los Rios  P., Dietler  G., Catsicas  S., Kasas  S.
ISSN
0027-8424 (Print)
Statut éditorial
Publié
Date de publication
07/2003
Volume
100
Numéro
15
Pages
8736-41
Notes
In Vitro
Journal Article
Research Support, Non-U.S. Gov't --- Old month value: Jul 22
Résumé
Measuring the biophysical properties of macromolecular complexes at work is a major challenge of modern biology. The protein complex composed of vesicle-associated membrane protein 2, synaptosomal-associated protein of 25 kDa, and syntaxin 1 [soluble N-ethyl-maleimide-sensitive factor attachment protein receptor (SNARE) complex] is essential for docking and fusion of neurotransmitter-filled synaptic vesicles with the presynaptic membrane. To better understand the fusion mechanisms, we reconstituted the synaptic SNARE complex in the imaging chamber of an atomic force microscope and measured the interaction forces between its components. Each protein was tested against the two others, taken either individually or as binary complexes. This approach allowed us to determine specific interaction forces and dissociation kinetics of the SNAREs and led us to propose a sequence of interactions. A theoretical model based on our measurements suggests that a minimum of four complexes is probably necessary for fusion to occur. We also showed that the regulatory protein neuronal Sec1 injected into the atomic force microscope chamber prevented the complex formation. Finally, we measured the effect of tetanus toxin protease on the SNARE complex and its activity by on-line registration during tetanus toxin injection. These experiments provide a basis for the functional study of protein microdomains and also suggest opportunities for sensitive screening of drugs that can modulate protein-protein interactions.
Mots-clé
Antigens, Surface/chemistry/physiology Biophysics Kinetics Macromolecular Substances Membrane Fusion/*physiology Membrane Proteins/chemistry/*physiology Microscopy, Atomic Force Munc18 Proteins Nerve Tissue Proteins/chemistry/pharmacology/physiology Protein Binding R-SNARE Proteins Recombinant Fusion Proteins/chemistry/metabolism SNARE Proteins Synaptic Vesicles/*physiology Synaptosomal-Associated Protein 25 Syntaxin 1 Tetanus Toxin/pharmacology Vesicular Transport Proteins/pharmacology/physiology
Pubmed
Web of science
Open Access
Oui
Création de la notice
24/01/2008 15:30
Dernière modification de la notice
20/08/2019 13:32
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