Clathrin coat controls synaptic vesicle acidification by blocking vacuolar ATPase activity

Farsi, ZohrehMax Planck Inst Biophys Chem, Dept Neurobiol, Gottingen, Germany.;Max Delbruck Ctr Mol Med, Berlin Inst Med Syst Biol, Berlin, Germany. (author)

QC 20180514
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.

Gowrisankaran, SindhujaUniv Med Ctr Gottingen, European Neurosci Inst, Synapt Vesicle Dynam Grp, Gottingen, Germany. (author)
Krunic, MatijaUniv Med Ctr Gottingen, European Neurosci Inst, Synapt Vesicle Dynam Grp, Gottingen, Germany. (author)
Rammner, BurkhardSciloop, Hamburg, Germany. (author)
Woehler, AndrewMax Delbruck Ctr Mol Med, Berlin Inst Med Syst Biol, Berlin, Germany. (author)
Lafer, Eileen M.Univ Texas Hlth Sci Ctr San Antonio, Dept Biochem & Struct Biol, San Antonio, TX 78229 USA.;Univ Texas Hlth Sci Ctr San Antonio, Ctr Biomed Neurosci, San Antonio, TX 78229 USA. (author)
Mim, CarstenKTH,Medicinteknik och hälsosystem(Swepub:kth)u1ce0qjf (author)
Jahn, ReinhardMax Planck Inst Biophys Chem, Dept Neurobiol, Gottingen, Germany. (author)
Milosevic, IraUniv Med Ctr Gottingen, European Neurosci Inst, Synapt Vesicle Dynam Grp, Gottingen, Germany. (author)
Max Planck Inst Biophys Chem, Dept Neurobiol, Gottingen, Germany.;Max Delbruck Ctr Mol Med, Berlin Inst Med Syst Biol, Berlin, Germany.Univ Med Ctr Gottingen, European Neurosci Inst, Synapt Vesicle Dynam Grp, Gottingen, Germany. (creator_code:org_t)

By the author/editor: Farsi, Zohreh; Gowrisankaran, S ...; Krunic, Matija; Rammner, Burkhar ...; Woehler, Andrew; Lafer, Eileen M.; show more...; Mim, Carsten; Jahn, Reinhard; Milosevic, Ira; show less...

About the subject

NATURAL SCIENCES: NATURAL SCIENCES; and Biological Scien ...; and Biophysics

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