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Yorodumi- EMDB-44842: Intact V-ATPase State 2 in synaptophysin knock-out isolated synap... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-44842 | |||||||||
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Title | Intact V-ATPase State 2 in synaptophysin knock-out isolated synaptic vesicles | |||||||||
Map data | Full map of V-ATPase State 2 in synaptophysin knock-out ISVs. | |||||||||
Sample |
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Keywords | V-ATPase / synaptic vesicle / MEMBRANE PROTEIN | |||||||||
Function / homology | Function and homology information Ion channel transport / Amino acids regulate mTORC1 / Transferrin endocytosis and recycling / Insulin receptor recycling / eye pigmentation / central nervous system maturation / Metabolism of Angiotensinogen to Angiotensins / transporter activator activity / negative regulation of autophagic cell death / plasma membrane proton-transporting V-type ATPase complex ...Ion channel transport / Amino acids regulate mTORC1 / Transferrin endocytosis and recycling / Insulin receptor recycling / eye pigmentation / central nervous system maturation / Metabolism of Angiotensinogen to Angiotensins / transporter activator activity / negative regulation of autophagic cell death / plasma membrane proton-transporting V-type ATPase complex / rostrocaudal neural tube patterning / RHOA GTPase cycle / positive regulation of transforming growth factor beta1 production / cellular response to increased oxygen levels / proton-transporting V-type ATPase, V1 domain / ROS and RNS production in phagocytes / proton-transporting V-type ATPase, V0 domain / synaptic vesicle lumen acidification / extrinsic component of synaptic vesicle membrane / P-type proton-exporting transporter activity / intracellular organelle / endosome to plasma membrane protein transport / lysosomal lumen acidification / clathrin-coated vesicle membrane / vacuolar transport / endosomal lumen acidification / vacuolar proton-transporting V-type ATPase, V0 domain / vacuolar proton-transporting V-type ATPase, V1 domain / vacuolar proton-transporting V-type ATPase complex / proton-transporting V-type ATPase complex / head morphogenesis / vacuolar acidification / osteoclast development / protein localization to cilium / dendritic spine membrane / regulation of cellular pH / ATPase complex / regulation of MAPK cascade / microvillus / ATPase activator activity / autophagosome membrane / positive regulation of Wnt signaling pathway / cilium assembly / transmembrane transporter complex / endomembrane system / regulation of macroautophagy / angiotensin maturation / ATP metabolic process / H+-transporting two-sector ATPase / axon terminus / ruffle / endoplasmic reticulum-Golgi intermediate compartment membrane / proton-transporting ATPase activity, rotational mechanism / RNA endonuclease activity / proton-transporting ATP synthase activity, rotational mechanism / Neutrophil degranulation / receptor-mediated endocytosis / proton transmembrane transport / secretory granule / transmembrane transport / cilium / small GTPase binding / synaptic vesicle membrane / endocytosis / positive regulation of canonical Wnt signaling pathway / melanosome / synaptic vesicle / apical part of cell / myelin sheath / signaling receptor activity / ATPase binding / cell body / postsynaptic membrane / intracellular iron ion homeostasis / Hydrolases; Acting on ester bonds / receptor-mediated endocytosis of virus by host cell / lysosome / positive regulation of ERK1 and ERK2 cascade / early endosome / endosome membrane / endosome / nuclear speck / apical plasma membrane / lysosomal membrane / axon / external side of plasma membrane / centrosome / ubiquitin protein ligase binding / endoplasmic reticulum membrane / protein-containing complex binding / perinuclear region of cytoplasm / Golgi apparatus / ATP hydrolysis activity / protein-containing complex / mitochondrion / extracellular space / nucleoplasm / ATP binding / membrane / plasma membrane Similarity search - Function | |||||||||
Biological species | Mus musculus (house mouse) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 4.4 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: 2024 Title: 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. | |||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_44842.map.gz | 321.4 MB | EMDB map data format | |
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Header (meta data) | emd-44842-v30.xml emd-44842.xml | 37.7 KB 37.7 KB | Display Display | EMDB header |
Images | emd_44842.png | 118.2 KB | ||
Filedesc metadata | emd-44842.cif.gz | 9.7 KB | ||
Others | emd_44842_additional_1.map.gz emd_44842_additional_2.map.gz emd_44842_half_map_1.map.gz emd_44842_half_map_2.map.gz | 318.4 MB 274.4 MB 274.1 MB 274.3 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-44842 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-44842 | HTTPS FTP |
-Validation report
Summary document | emd_44842_validation.pdf.gz | 1.2 MB | Display | EMDB validaton report |
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Full document | emd_44842_full_validation.pdf.gz | 1.2 MB | Display | |
Data in XML | emd_44842_validation.xml.gz | 16.6 KB | Display | |
Data in CIF | emd_44842_validation.cif.gz | 19.5 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-44842 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-44842 | HTTPS FTP |
-Related structure data
Related structure data | 9brsMC 9braC 9brqC 9brrC 9brtC 9bruC 9bryC 9brzC M: atomic model generated by this map C: citing same article (ref.) |
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Similar structure data | Similarity search - Function & homologyF&H Search |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_44842.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 2 in synaptophysin knock-out ISVs. | ||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.1064 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 2 in synaptophysin knock-out ISVs.
File | emd_44842_additional_1.map | ||||||||||||
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Annotation | EMhancer map of V-ATPase State 2 in synaptophysin knock-out ISVs. | ||||||||||||
Projections & Slices |
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Density Histograms |
-Additional map: Unsharpened map
File | emd_44842_additional_2.map | ||||||||||||
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Annotation | Unsharpened map | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Half map of V-ATPase State 2 in synaptophysin knock-out ISVs.
File | emd_44842_half_map_1.map | ||||||||||||
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Annotation | Half map of V-ATPase State 2 in synaptophysin knock-out ISVs. | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Half map of V-ATPase State 2 in synaptophysin knock-out ISVs.
File | emd_44842_half_map_2.map | ||||||||||||
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Annotation | Half map of V-ATPase State 2 in synaptophysin knock-out ISVs. | ||||||||||||
Projections & Slices |
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Density Histograms |
-Sample components
+Entire : Synaptophysin knock-out mouse brain isolated glutamatergic synapt...
+Supramolecule #1: Synaptophysin knock-out mouse brain isolated glutamatergic synapt...
+Macromolecule #1: V-type proton ATPase subunit E 1
+Macromolecule #2: V-type proton ATPase subunit G 2
+Macromolecule #3: V-type proton ATPase subunit C 1
+Macromolecule #4: V-type proton ATPase subunit H
+Macromolecule #5: V-type proton ATPase 116 kDa subunit a 1
+Macromolecule #6: V-type proton ATPase subunit e 2
+Macromolecule #7: Ribonuclease kappa
+Macromolecule #8: V-type proton ATPase subunit F
+Macromolecule #9: V-type proton ATPase subunit d 1
+Macromolecule #10: V-type proton ATPase subunit S1
+Macromolecule #11: Renin receptor cytoplasmic fragment
+Macromolecule #12: V-type proton ATPase 21 kDa proteolipid subunit c''
+Macromolecule #13: V-type proton ATPase 16 kDa proteolipid subunit c
+Macromolecule #14: V-type proton ATPase catalytic subunit A
+Macromolecule #15: V-type proton ATPase subunit B, brain isoform
+Macromolecule #16: V-type proton ATPase subunit D
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | cell |
-Sample preparation
Buffer | pH: 7.4 |
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Vitrification | Cryogen name: ETHANE |
Details | The specimen state should be an intact subcellular component. |
-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: 2.0 µm / Nominal defocus min: 0.8 µm |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
-Image processing
Startup model | Type of model: OTHER |
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Final reconstruction | Resolution.type: BY AUTHOR / Resolution: 4.4 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 30487 |
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-9brs: |