High-resolution electron cryomicroscopy of V-ATPase in native synaptic vesicles

Intercellular communication in the nervous system occurs through the release of neurotransmitters into the synaptic cleft between neurons. In the presynaptic neuron, the proton pumping vesicular- or vacuolar-type ATPase (V-ATPase) powers neurotransmitter loading into synaptic vesicles (SVs), with th...

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Published in:Science (American Association for the Advancement of Science) 2024-07, Vol.385 (6705), p.168-174
Main Authors: Coupland, Claire E., Karimi, Ryan, Bueler, Stephanie A., Liang, Yingke, Courbon, Gautier M., Di Trani, Justin M., Wong, Cassandra J., Saghian, Rayan, Youn, Ji-Young, Wang, Lu-Yang, Rubinstein, John L.
Format: Article
Language:English
Subjects:
Adenosine triphosphatase
Chemical communication
Cytosol
H+-transporting ATPase
High resolution
Membrane proteins
Membrane vesicles
Neurotransmitters
Protons
Synaptic vesicles
Synaptophysin
Vesicles
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Summary:Intercellular communication in the nervous system occurs through the release of neurotransmitters into the synaptic cleft between neurons. In the presynaptic neuron, the proton pumping vesicular- or vacuolar-type ATPase (V-ATPase) powers neurotransmitter loading into synaptic vesicles (SVs), with the V 1 complex dissociating from the membrane region of the enzyme before exocytosis. We isolated SVs from rat brain using SidK, a V-ATPase–binding bacterial effector protein. Single-particle electron cryomicroscopy allowed high-resolution structure determination of V-ATPase within the native SV membrane. In the structure, regularly spaced cholesterol molecules decorate the enzyme’s rotor and the abundant SV protein synaptophysin binds the complex stoichiometrically. ATP hydrolysis during vesicle loading results in a loss of the V 1 region of V-ATPase from the SV membrane, suggesting that loading is sufficient to induce dissociation of the enzyme. Editor’s summary Chemical communication between neurons involves the rapid release of neurotransmitters into the synapse. Vesicular-type ATPase (V-ATPase) uses ATP to pump protons into synaptic vesicles, enabling neurotransmitter uptake from the cytosol before release. Coupland et al . isolated whole synaptic vesicles from rat brain and were able to determine high-resolution electron cryomicroscopy structures of the V-ATPase in its native membrane. The authors found several unexpected features that were not seen in structures of purified V-ATPase, including a stoichiometric complex with the protein synaptophysin. They also observed dissociation of the V1 head group under active conditions, consistent with prior biochemical work. —Michael A. Funk
ISSN:0036-8075
1095-9203
1095-9203
DOI:10.1126/science.adp5577