Involvement of Ca2+ channel synprint site in synaptic vesicle endocytosis

Hiroyasu Watanabe; Takayuki Yamashita; Naoto Saitoh; Shigeki Kiyonaka; Akihiro Iwamatsu; Kevin P Campbell; Yasuo Mori; Tomoyuki Takahashi

doi:10.1523/JNEUROSCI.3214-09.2010

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Involvement of Ca2+ channel synprint site in synaptic vesicle endocytosis

Journal article

Open access

Peer reviewed

Involvement of Ca2+ channel synprint site in synaptic vesicle endocytosis

Hiroyasu Watanabe, Takayuki Yamashita, Naoto Saitoh, Shigeki Kiyonaka, Akihiro Iwamatsu, Kevin P Campbell, Yasuo Mori and Tomoyuki Takahashi

The Journal of neuroscience, Vol.30(2), pp.655-660

01/13/2010

DOI: 10.1523/JNEUROSCI.3214-09.2010

PMID: 20071530

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Abstract

The synaptic protein interaction (synprint) site of the voltage-gated Ca(2+) channel (VGCC) alpha1 subunit can interact with proteins involved in exocytosis, and it is therefore thought to be essential for exocytosis of synaptic vesicles. Here we report that the synprint site can also directly bind the mu subunit of AP-2, an adaptor protein for clathrin-mediated endocytosis, in competition with the synaptotagmin 1 (Syt 1) C2B domain. In brain lysates, the AP-2-synprint interaction occurred over a wide range of Ca(2+) concentrations but was inhibited at high Ca(2+) concentrations, in which Syt 1 interacted with synprint site. At the calyx of Held synapse in rat brainstem slices, direct presynaptic loading of the synprint fragment peptide blocked endocytic, but not exocytic, membrane capacitance changes. We propose that the VGCC synprint site is involved in synaptic vesicle endocytosis, rather than exocytosis, in the nerve terminal, via Ca(2+)-dependent interactions with AP-2 and Syt.

Calcium Channels - cerebrospinal fluid

Protein Binding - genetics

Calcium Channels - metabolism

Rats, Wistar

Calcium - metabolism

Biophysical Phenomena - drug effects

Green Fluorescent Proteins - genetics

Structure-Activity Relationship

Recombinant Fusion Proteins

DNA-Binding Proteins - metabolism

Dose-Response Relationship, Drug

Synapses - metabolism

Mass Spectrometry

Binding Sites - physiology

Neurons - physiology

Biophysical Phenomena - genetics

Neurons - drug effects

Calcium Channels - genetics

Electric Stimulation - methods

Synaptic Vesicles - physiology

Animals, Newborn

Membrane Potentials - drug effects

Calcium - pharmacology

Myelin Basic Protein - metabolism

Point Mutation - physiology

Rats

Membrane Potentials - physiology

Endocytosis - physiology

Patch-Clamp Techniques

Animals

Synaptotagmin I - metabolism

Superior Colliculi - cytology

Presynaptic Terminals - metabolism

In Vitro Techniques

Details

Title: Subtitle: Involvement of Ca2+ channel synprint site in synaptic vesicle endocytosis
Creators: Hiroyasu Watanabe - Cellular and Molecular Synaptic Function Unit, Initial Research Project, Okinawa Institute of Science and Technology Promotion Corporation, Okinawa 904-2234, Japan
Takayuki Yamashita
Naoto Saitoh
Shigeki Kiyonaka
Akihiro Iwamatsu
Kevin P Campbell
Yasuo Mori
Tomoyuki Takahashi
Resource Type: Journal article
Publication Details: The Journal of neuroscience, Vol.30(2), pp.655-660
Publisher: United States
DOI: 10.1523/JNEUROSCI.3214-09.2010
PMID: 20071530
ISSN: 0270-6474
eISSN: 1529-2401
Language: English
Date published: 01/13/2010
Academic Unit: Neurology; Molecular Physiology and Biophysics; Iowa Neuroscience Institute
Record Identifier: 9984020884702771

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