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PMID: 11222632 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

synaptotagmin mutants reveal essential functions for the C2B domain in Ca2+-triggered fusion and recycling of synaptic vesicles in vivo.

The Journal of neuroscience : the official journal of the Society for Neuroscience ·Vol. 21 ·No. 5 ·2001-03-01 ·Pages 1421-33

Littleton JT, Bai J, Vyas B, Desai R, Baltus AE, Garment MB, Carlson SD, Ganetzky B, Chapman ER

Abstract

Synaptotagmin has been proposed to function as a Ca(2+) sensor that regulates synaptic vesicle exocytosis, whereas the soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complex is thought to form the core of a conserved membrane fusion machine. Little is known concerning the functional relationships between synaptotagmin and SNAREs. Here we report that synaptotagmin can facilitate SNARE complex formation in vitro and that synaptotagmin mutations disrupt SNARE complex formation in vivo. Synaptotagmin oligomers efficiently bind SNARE complexes, whereas Ca(2+) acting via synaptotagmin triggers cross-linking of SNARE complexes into dimers. Mutations in Drosophila that delete the C2B domain of synaptotagmin disrupt clathrin AP-2 binding and endocytosis. In contrast, a mutation that blocks Ca(2+)-triggered conformational changes in C2B and diminishes Ca(2+)-triggered synaptotagmin oligomerization results in a postdocking defect in neurotransmitter release and a decrease in SNARE assembly in vivo. These data suggest that Ca(2+)-driven oligomerization via the C2B domain of synaptotagmin may trigger synaptic vesicle fusion via the assembly and clustering of SNARE complexes.

MeSH Terms
Adaptor Protein Complex alpha Subunits Adaptor Proteins, Vesicular Transport Animals Biopolymers/biosynthesis,chemistry Calcium/metabolism,pharmacology Calcium-Binding Proteins Dimerization Drosophila Endocytosis/physiology Exocytosis/physiology Macromolecular Substances Membrane Fusion/drug effects,physiology Membrane Glycoproteins/chemistry,genetics,metabolism Membrane Proteins/chemistry,metabolism Mutation Nerve Tissue Proteins/chemistry,genetics,metabolism Precipitin Tests Protein Conformation Protein Structure, Tertiary/genetics Rats SNARE Proteins Structure-Activity Relationship Synaptic Vesicles/metabolism Synaptotagmins Vesicular Transport Proteins
Chemicals
Adaptor Protein Complex alpha Subunits Adaptor Proteins, Vesicular Transport Biopolymers Calcium-Binding Proteins Macromolecular Substances Membrane Glycoproteins Membrane Proteins Nerve Tissue Proteins SNARE Proteins Vesicular Transport Proteins Synaptotagmins Calcium
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Littleton J T
Department of Physiology and Entomology, University of Wisconsin, Madison, Wisconsin 53706, USA.
Bai J
Vyas B
Desai R
Baltus A E
Garment M B
Carlson S D
Ganetzky B
Chapman E R
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Article Info
Journal
The Journal of neuroscience : the official journal of the Society for Neuroscience
Abbr.
J Neurosci
ISSN
1529-2401
Published
2001-03-01
Pages
1421-33
Language
English
Region
United States
NLM ID
8102140
PMCID
PMC6762938
Subset
IM
Grants
NIGMS NIH HHS · GM43100 · United States
NINDS NIH HHS · NS15390 · United States
NINDS NIH HHS · R01 NS015390 · United States
NIGMS NIH HHS · GM 56827-01 · United States
NIGMS NIH HHS · R01 GM056827 · United States
NINDS NIH HHS · NS40296-01 · United States
NINDS NIH HHS · R01 NS040296 · United States
NINDS NIH HHS · R01 NS040296-01 · United States
External Links
PubMed source PMC Full Text View journal details
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