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Yorodumi- EMDB-44840: Intact V-ATPase State 3 and synaptophysin complex in mouse brain ... -
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Basic information
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| Title | Intact V-ATPase State 3 and synaptophysin complex in mouse brain isolated synaptic vesicles | |||||||||
Map data | Full map of V-ATPase State 3 synaptophysin complex in wild-type ISVs. | |||||||||
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Keywords | V-ATPase / synaptic vesicle / MEMBRANE PROTEIN | |||||||||
| Function / homology | Function and homology informationIon channel transport / regulation of opioid receptor signaling pathway / Amino acids regulate mTORC1 / clathrin-sculpted glutamate transport vesicle membrane / Transferrin endocytosis and recycling / Insulin receptor recycling / Metabolism of Angiotensinogen to Angiotensins / eye pigmentation / central nervous system maturation / rostrocaudal neural tube patterning ...Ion channel transport / regulation of opioid receptor signaling pathway / Amino acids regulate mTORC1 / clathrin-sculpted glutamate transport vesicle membrane / Transferrin endocytosis and recycling / Insulin receptor recycling / Metabolism of Angiotensinogen to Angiotensins / eye pigmentation / central nervous system maturation / rostrocaudal neural tube patterning / proton-transporting V-type ATPase, V1 domain / ROS and RNS production in phagocytes / positive regulation of transforming growth factor beta1 production / RHOA GTPase cycle / synaptic vesicle lumen acidification / P-type proton-exporting transporter activity / regulation of synaptic vesicle priming / proton-transporting V-type ATPase, V0 domain / extrinsic component of synaptic vesicle membrane / cellular response to increased oxygen levels / vacuolar proton-transporting V-type ATPase, V1 domain / endosome to plasma membrane protein transport / vacuolar proton-transporting V-type ATPase, V0 domain / clathrin-coated vesicle membrane / lysosomal lumen acidification / endosomal lumen acidification / proton-transporting V-type ATPase complex / head morphogenesis / protein localization to cilium / vacuolar proton-transporting V-type ATPase complex / osteoclast development / neuron projection terminus / regulation of short-term neuronal synaptic plasticity / vacuolar acidification / dendritic spine membrane / presynaptic active zone / syntaxin-1 binding / cholesterol binding / ATPase complex / microvillus / ATPase activator activity / regulation of MAPK cascade / autophagosome membrane / excitatory synapse / proton-transporting ATPase activity, rotational mechanism / cilium assembly / positive regulation of Wnt signaling pathway / regulation of macroautophagy / transporter activator activity / H+-transporting two-sector ATPase / ATP metabolic process / angiotensin maturation / ruffle / receptor-mediated endocytosis of virus by host cell / Neutrophil degranulation / axon terminus / endoplasmic reticulum-Golgi intermediate compartment membrane / endomembrane system / RNA endonuclease activity / SH2 domain binding / proton transmembrane transport / receptor-mediated endocytosis / secretory granule / neuromuscular junction / regulation of long-term neuronal synaptic plasticity / modulation of chemical synaptic transmission / small GTPase binding / Schaffer collateral - CA1 synapse / endocytosis / terminal bouton / apical part of cell / synaptic vesicle / synaptic vesicle membrane / melanosome / positive regulation of canonical Wnt signaling pathway / myelin sheath / presynapse / signaling receptor activity / presynaptic membrane / ATPase binding / chemical synaptic transmission / Hydrolases; Acting on ester bonds / intracellular iron ion homeostasis / postsynaptic membrane / early endosome / lysosome / endosome / neuron projection / endosome membrane / nuclear speck / cilium / apical plasma membrane / axon / lysosomal membrane / external side of plasma membrane / synapse / ubiquitin protein ligase binding / centrosome / endoplasmic reticulum membrane / protein-containing complex binding Similarity search - Function | |||||||||
| Biological species | ![]() | |||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 4.3 Å | |||||||||
Authors | Wang C / Jiang W / Yang K / Wang X / Guo Q / Brunger AT | |||||||||
| Funding support | United States, 2 items
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Citation | Journal: Nature / Year: 2024Title: Structure and topography of the synaptic V-ATPase-synaptophysin complex. Authors: Chuchu Wang / Wenhong Jiang / Jeremy Leitz / Kailu Yang / Luis Esquivies / Xing Wang / Xiaotao Shen / Richard G Held / Daniel J Adams / Tamara Basta / Lucas Hampton / Ruiqi Jian / Lihua ...Authors: Chuchu Wang / Wenhong Jiang / Jeremy Leitz / Kailu Yang / Luis Esquivies / Xing Wang / Xiaotao Shen / Richard G Held / Daniel J Adams / Tamara Basta / Lucas Hampton / Ruiqi Jian / Lihua Jiang / Michael H B Stowell / Wolfgang Baumeister / Qiang Guo / Axel T Brunger / ![]() Abstract: Synaptic vesicles are organelles with a precisely defined protein and lipid composition, yet the molecular mechanisms for the biogenesis of synaptic vesicles are mainly unknown. Here we discovered a ...Synaptic vesicles are organelles with a precisely defined protein and lipid composition, yet the molecular mechanisms for the biogenesis of synaptic vesicles are mainly unknown. Here we discovered a well-defined interface between the synaptic vesicle V-ATPase and synaptophysin by in situ cryo-electron tomography and single-particle cryo-electron microscopy of functional synaptic vesicles isolated from mouse brains. The synaptic vesicle V-ATPase is an ATP-dependent proton pump that establishes the proton gradient across the synaptic vesicle, which in turn drives the uptake of neurotransmitters. Synaptophysin and its paralogues synaptoporin and synaptogyrin belong to a family of abundant synaptic vesicle proteins whose function is still unclear. We performed structural and functional studies of synaptophysin-knockout mice, confirming the identity of synaptophysin as an interaction partner with the V-ATPase. Although there is little change in the conformation of the V-ATPase upon interaction with synaptophysin, the presence of synaptophysin in synaptic vesicles profoundly affects the copy number of V-ATPases. This effect on the topography of synaptic vesicles suggests that synaptophysin assists in their biogenesis. In support of this model, we observed that synaptophysin-knockout mice exhibit severe seizure susceptibility, suggesting an imbalance of neurotransmitter release as a physiological consequence of the absence of synaptophysin. | |||||||||
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Structure visualization
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Downloads & links
-EMDB archive
| Map data | emd_44840.map.gz | 322 MB | EMDB map data format | |
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| Header (meta data) | emd-44840-v30.xml emd-44840.xml | 38.9 KB 38.9 KB | Display Display | EMDB header |
| Images | emd_44840.png | 152.9 KB | ||
| Filedesc metadata | emd-44840.cif.gz | 9.9 KB | ||
| Others | emd_44840_additional_1.map.gz emd_44840_additional_2.map.gz emd_44840_half_map_1.map.gz emd_44840_half_map_2.map.gz | 319.3 MB 273.8 MB 318 MB 318 MB | ||
| Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-44840 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-44840 | HTTPS FTP |
-Validation report
| Summary document | emd_44840_validation.pdf.gz | 1.1 MB | Display | EMDB validaton report |
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| Full document | emd_44840_full_validation.pdf.gz | 1.1 MB | Display | |
| Data in XML | emd_44840_validation.xml.gz | 16.9 KB | Display | |
| Data in CIF | emd_44840_validation.cif.gz | 19.9 KB | Display | |
| Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-44840 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-44840 | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 9brqMC ![]() 9braC ![]() 9brrC ![]() 9brsC ![]() 9brtC ![]() 9bruC ![]() 9bryC ![]() 9brzC M: atomic model generated by this map C: citing same article ( |
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| Similar structure data | Similarity search - Function & homology F&H Search |
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Links
| EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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| Related items in Molecule of the Month |
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Map
| File | Download / File: emd_44840.map.gz / Format: CCP4 / Size: 343 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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| Annotation | Full map of V-ATPase State 3 synaptophysin complex in wild-type ISVs. | ||||||||||||||||||||||||||||||||||||
| Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
| Voxel size | X=Y=Z: 1.11138 Å | ||||||||||||||||||||||||||||||||||||
| Density |
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| Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
| Details | EMDB XML:
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-Supplemental data
-Additional map: EMhancer map of V-ATPase State 3 synaptophysin complex...
| File | emd_44840_additional_1.map | ||||||||||||
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| Annotation | EMhancer map of V-ATPase State 3 synaptophysin complex in wild-type ISVs. | ||||||||||||
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| Density Histograms |
-Additional map: Unsharpened map of V-ATPase State 3
| File | emd_44840_additional_2.map | ||||||||||||
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| Annotation | Unsharpened map of V-ATPase State 3 | ||||||||||||
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| Density Histograms |
-Half map: Half map of V-ATPase State 3 synaptophysin complex in wild-type ISVs.
| File | emd_44840_half_map_1.map | ||||||||||||
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| Annotation | Half map of V-ATPase State 3 synaptophysin complex in wild-type ISVs. | ||||||||||||
| Projections & Slices |
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| Density Histograms |
-Half map: Half map of V-ATPase State 3 synaptophysin complex in wild-type ISVs.
| File | emd_44840_half_map_2.map | ||||||||||||
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| Annotation | Half map of V-ATPase State 3 synaptophysin complex in wild-type ISVs. | ||||||||||||
| Projections & Slices |
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| Density Histograms |
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Sample components
+Entire : Mouse brain isolated glutamatergic synaptic vesicles
+Supramolecule #1: Mouse brain isolated glutamatergic synaptic vesicles
+Macromolecule #1: V-type proton ATPase 21 kDa proteolipid subunit c''
+Macromolecule #2: V-type proton ATPase 16 kDa proteolipid subunit c
+Macromolecule #3: V-type proton ATPase catalytic subunit A
+Macromolecule #4: V-type proton ATPase subunit B, brain isoform
+Macromolecule #5: V-type proton ATPase subunit D
+Macromolecule #6: V-type proton ATPase subunit E 1
+Macromolecule #7: V-type proton ATPase subunit G 2
+Macromolecule #8: V-type proton ATPase subunit F
+Macromolecule #9: V-type proton ATPase subunit C 1
+Macromolecule #10: V-type proton ATPase subunit H
+Macromolecule #11: V-type proton ATPase subunit d 1
+Macromolecule #12: V-type proton ATPase 116 kDa subunit a 1
+Macromolecule #13: V-type proton ATPase subunit S1
+Macromolecule #14: Renin receptor cytoplasmic fragment
+Macromolecule #15: Ribonuclease kappa
+Macromolecule #16: Synaptophysin
+Macromolecule #17: V-type proton ATPase subunit e 2
-Experimental details
-Structure determination
| Method | cryo EM |
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Processing | single particle reconstruction |
| Aggregation state | cell |
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Sample preparation
| Buffer | pH: 7.4 |
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| Vitrification | Cryogen name: ETHANE |
| Details | The specimen state should be an intact subcellular component. |
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Electron microscopy
| Microscope | TFS KRIOS |
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| Image recording | Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Average electron dose: 50.0 e/Å2 |
| Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
| Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 4.0 µm / Nominal defocus min: 1.0 µm |
| Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
| Startup model | Type of model: OTHER |
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| Final reconstruction | Resolution.type: BY AUTHOR / Resolution: 4.3 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 34536 |
| Initial angle assignment | Type: RANDOM ASSIGNMENT |
| Final angle assignment | Type: MAXIMUM LIKELIHOOD |
-Atomic model buiding 1
| Refinement | Protocol: FLEXIBLE FIT |
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| Output model | ![]() PDB-9brq: |
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United States, 2 items
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FIELD EMISSION GUN
