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PMID: 15456828 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Dual roles of the C2B domain of synaptotagmin I in synchronizing Ca2+-dependent neurotransmitter release.

The Journal of neuroscience : the official journal of the Society for Neuroscience ·Vol. 24 ·No. 39 ·2004-09-29 ·Pages 8542-50

Nishiki T, Augustine GJ

Abstract

Although the vesicular protein synaptotagmin I contains two Ca2+-binding domains (C2A and C2B), Ca2+ binding to the C2B domain is more important for triggering synchronous neurotransmitter release. We have used point mutagenesis to determine the functional contributions of the five negatively charged aspartate (Asp) residues that constitute the Ca2+-binding sites in the C2B domain of synaptotagmin I. Transfecting wild-type synaptotagmin I DNA into cultured hippocampal neurons from synaptotagmin I knock-out mice rescued Ca2+-dependent synchronous transmitter release and reduced a slower, asynchronous component of release, indicating that synaptotagmin I suppresses asynchronous release. Mutating either the second or third Asp residues of the C2B domain potently inhibited the ability of synaptotagmin I to rescue synchronous release but did not change its ability to suppress asynchronous release. Synaptotagmin I with mutations in the first or fourth Asp residues of the C2B domain partially rescued synchronous release and partially suppressed asynchronous release, whereas neutralizing the fifth Asp residue had no effect on the ability of synaptotagmin I to rescue transmitter release. Thus, we conclude that the C2B domain of synaptotagmin I regulates neurotransmitter release in at least two ways. Synchronous release absolutely requires binding of Ca2+ to the second and third Asp residues in this domain. For the suppression of asynchronous release, Ca2+ binding to the C2B domain of synaptotagmin I apparently is not necessary because mutation of the second Asp residue inhibits Ca2+ binding, yet still allows this protein to suppress asynchronous release.

MeSH Terms
Animals Aspartic Acid Binding Sites/genetics Calcium/metabolism,physiology Calcium-Binding Proteins/genetics,physiology Cells, Cultured Excitatory Postsynaptic Potentials/physiology Hippocampus/cytology,metabolism Membrane Glycoproteins/genetics,physiology Mice Mice, Knockout Mutagenesis, Site-Directed Nerve Tissue Proteins/genetics,physiology Neurons/metabolism Neurotransmitter Agents/metabolism Protein Structure, Tertiary Synaptic Vesicles/metabolism Synaptotagmin I Synaptotagmins Time Factors Transfection
Chemicals
Calcium-Binding Proteins Membrane Glycoproteins Nerve Tissue Proteins Neurotransmitter Agents Synaptotagmin I Syt1 protein, mouse Synaptotagmins Aspartic Acid Calcium
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Nishiki Tei-ichi
Department of Neurobiology, Duke University Medical Center, Durham, North Carolina 27710, USA.
Augustine George J
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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
DOI
10.1523/JNEUROSCI.2545-04.2004
Published
2004-09-29
Pages
8542-50
Language
English
Region
United States
NLM ID
8102140
PMCID
PMC6729890
Subset
IM
Grants
NIMH NIH HHS · R01 MH067044 · United States
NINDS NIH HHS · R01 NS021624 · United States
NIMH NIH HHS · MH-67044 · United States
NINDS NIH HHS · NS-21624 · United States
External Links
PubMed source DOI: 10.1523/JNEUROSCI.2545-04.2004 PMC Full Text View journal details
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