Synaptophysin controls synaptobrevin-II retrieval via a cryptic C-terminal interaction site.
Endocytosis
/ genetics
Gene Knockout Techniques
Hippocampus
/ metabolism
Neurons
/ chemistry
Primary Cell Culture
SNARE Proteins
/ genetics
Synaptic Transmission
/ genetics
Synaptic Vesicles
/ genetics
Synaptophysin
/ chemistry
Synaptosomes
/ chemistry
Vesicle-Associated Membrane Protein 2
/ genetics
endocytosis
exocytosis
neuron
neurotransmitter release
synapse
synaptosome
vesicles
Journal
The Journal of biological chemistry
ISSN: 1083-351X
Titre abrégé: J Biol Chem
Pays: United States
ID NLM: 2985121R
Informations de publication
Date de publication:
Historique:
received:
17
03
2020
revised:
03
01
2021
accepted:
06
01
2021
pubmed:
27
3
2021
medline:
26
8
2021
entrez:
26
3
2021
Statut:
ppublish
Résumé
The accurate retrieval of synaptic vesicle (SV) proteins during endocytosis is essential for the maintenance of neurotransmission. Synaptophysin (Syp) and synaptobrevin-II (SybII) are the most abundant proteins on SVs. Neurons lacking Syp display defects in the activity-dependent retrieval of SybII and a general slowing of SV endocytosis. To determine the role of the cytoplasmic C terminus of Syp in the control of these two events, we performed molecular replacement studies in primary cultures of Syp knockout neurons using genetically encoded reporters of SV cargo trafficking at physiological temperatures. Under these conditions, we discovered, 1) no slowing in SV endocytosis in Syp knockout neurons, and 2) a continued defect in SybII retrieval in knockout neurons expressing a form of Syp lacking its C terminus. Sequential truncations of the Syp C-terminus revealed a cryptic interaction site for the SNARE motif of SybII that was concealed in the full-length form. This suggests that a conformational change within the Syp C terminus is key to permitting SybII binding and thus its accurate retrieval. Furthermore, this study reveals that the sole presynaptic role of Syp is the control of SybII retrieval, since no defect in SV endocytosis kinetics was observed at physiological temperatures.
Identifiants
pubmed: 33769286
pii: S0021-9258(21)00033-8
doi: 10.1016/j.jbc.2021.100266
pmc: PMC7948965
pii:
doi:
Substances chimiques
SNARE Proteins
0
Synaptophysin
0
Vesicle-Associated Membrane Protein 2
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
100266Subventions
Organisme : Wellcome Trust
Pays : United Kingdom
Organisme : Wellcome Trust
ID : 204954/Z/16/Z
Pays : United Kingdom
Informations de copyright
Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.
Déclaration de conflit d'intérêts
Conflicts of interest The authors declare that they have no conflicts of interest with the contents of this article.
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