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TitleHigh-resolution electron cryomicroscopy of V-ATPase in native synaptic vesicles.
Journal, issue, pagesScience, Vol. 385, Issue 6705, Page 168-174, Year 2024
Publish dateJul 12, 2024
AuthorsClaire E Coupland / Ryan Karimi / Stephanie A Bueler / Yingke Liang / Gautier M Courbon / Justin M Di Trani / Cassandra J Wong / Rayan Saghian / Ji-Young Youn / Lu-Yang Wang / John L Rubinstein /
PubMed AbstractIntercellular 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 ...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 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 region of V-ATPase from the SV membrane, suggesting that loading is sufficient to induce dissociation of the enzyme.
External linksScience / PubMed:38900912
MethodsEM (single particle)
Resolution3.2 - 3.9 Å
Structure data

44350

9b8o

EMDB-44350, PDB-9b8o:
Synaptic Vesicle V-ATPase with synaptophysin and SidK, State 3, Vo
Method: EM (single particle) / Resolution: 3.2 Å

44351

9b8p

EMDB-44351, PDB-9b8p:
Synaptic Vesicle V-ATPase with synaptophysin and SidK, State 3, V1
Method: EM (single particle) / Resolution: 3.2 Å

44352

9b8q

EMDB-44352, PDB-9b8q:
Synaptic Vesicle V-ATPase with synaptophysin and SidK, State 3, peripheral stalks
Method: EM (single particle) / Resolution: 3.8 Å

44353

9brd

EMDB-44353, PDB-9brd:
Synaptic Vesicle V-ATPase with synaptophysin and SidK, State 3
Method: EM (single particle) / Resolution: 3.5 Å

44354

9brc

EMDB-44354, PDB-9brc:
Synaptic Vesicle V-ATPase with synaptophysin and SidK, State 2
Method: EM (single particle) / Resolution: 3.9 Å

44355

9brb

EMDB-44355, PDB-9brb:
Synaptic Vesicle V-ATPase with synaptophysin and SidK, State 1
Method: EM (single particle) / Resolution: 3.6 Å

Chemicals

ChemComp-PC1:
1,2-DIACYL-SN-GLYCERO-3-PHOSPHOCHOLINE / phospholipid*YM

ChemComp-WJP:
methyl (3R,6Z,10E,14E)-3,7,11,15,19-pentamethylicosa-6,10,14,18-tetraen-1-yl dihydrogen diphosphate

ChemComp-NAG:
2-acetamido-2-deoxy-beta-D-glucopyranose

ChemComp-LP3:
(7R)-4,7-DIHYDROXY-N,N,N-TRIMETHYL-10-OXO-3,5,9-TRIOXA-4-PHOSPHAHEPTACOSAN-1-AMINIUM 4-OXIDE

ChemComp-PTY:
PHOSPHATIDYLETHANOLAMINE / phospholipid*YM

ChemComp-CLR:
CHOLESTEROL

ChemComp-ADP:
ADENOSINE-5'-DIPHOSPHATE / ADP, energy-carrying molecule*YM

Source
  • rattus norvegicus (Norway rat)
  • legionella pneumophila subsp. pneumophila str. philadelphia 1 (bacteria)
  • legionella pneumophila subsp. pneumophila (bacteria)
KeywordsPROTON TRANSPORT / Membrane / Synaptic / Complex / Mmebrane / Native / Vesicle

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