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Elife. 2018 Apr 13;7. pii: e32569. doi: 10.7554/eLife.32569.

Clathrin coat controls synaptic vesicle acidification by blocking vacuolar ATPase activity.

Author information

1
Department of Neurobiology, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany.
2
Berlin Institute for Medical Systems Biology, Max Delbrück Center for Molecular Medicine, Berlin, Germany.
3
Synaptic Vesicle Dynamics Group, European Neuroscience Institute, University Medical Center Göttingen, Göttingen, Germany.
4
Sciloop, Hamburg, Germany.
5
Department of Biochemistry and Structural Biology, University of Texas Health Science Center at San Antonio, San Antonio, United States.
6
Center for Biomedical Neuroscience, University of Texas Health Science Center at San Antonio, San Antonio, United States.
7
Department for Biomedical Engineering and Health Solutions, Kungliga Tekniska Högskolan, Huddinge, Sweden.
8
Department of Biosciences and Nutrition, Karolinska Institute, Huddinge, Sweden.
#
Contributed equally

Abstract

Newly-formed synaptic vesicles (SVs) are rapidly acidified by vacuolar adenosine triphosphatases (vATPases), generating a proton electrochemical gradient that drives neurotransmitter loading. Clathrin-mediated endocytosis is needed for the formation of new SVs, yet it is unclear when endocytosed vesicles acidify and refill at the synapse. Here, we isolated clathrin-coated vesicles (CCVs) from mouse brain to measure their acidification directly at the single vesicle level. We observed that the ATP-induced acidification of CCVs was strikingly reduced in comparison to SVs. Remarkably, when the coat was removed from CCVs, uncoated vesicles regained ATP-dependent acidification, demonstrating that CCVs contain the functional vATPase, yet its function is inhibited by the clathrin coat. Considering the known structures of the vATPase and clathrin coat, we propose a model in which the formation of the coat surrounds the vATPase and blocks its activity. Such inhibition is likely fundamental for the proper timing of SV refilling.

KEYWORDS:

E. coli; acidification; clathrin coat; endocytosis; human; mouse; neuroscience; proton pump; synaptic vesicle; vATPase

PMID:
29652249
PMCID:
PMC5935483
DOI:
10.7554/eLife.32569
[Indexed for MEDLINE]
Free PMC Article

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