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- EMDB-25666: Rat vesicular glutamate transporter 2 (VGLUT2) in low Cl condition -

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Basic information

Entry
Database: EMDB / ID: EMD-25666
TitleRat vesicular glutamate transporter 2 (VGLUT2) in low Cl condition
Map datasharpened map for model building
Sample
  • Complex: complex of WT VGLUT2 with a Fab
    • Protein or peptide: Vesicular glutamate transporter 2
Keywordssynaptic vesicle / neurotransmitter / glutamate transport / TRANSPORT PROTEIN
Function / homology
Function and homology information


sodium:phosphate symporter activity / sodium-dependent phosphate transport / phosphate ion uniporter activity / L-glutamate uniporter activity / pericellular basket / Organic anion transporters / neurotransmitter uptake / neurotransmitter loading into synaptic vesicle / hyaloid vascular plexus regression / L-glutamate import ...sodium:phosphate symporter activity / sodium-dependent phosphate transport / phosphate ion uniporter activity / L-glutamate uniporter activity / pericellular basket / Organic anion transporters / neurotransmitter uptake / neurotransmitter loading into synaptic vesicle / hyaloid vascular plexus regression / L-glutamate import / phosphate ion transport / L-glutamate transmembrane transport / L-glutamate transmembrane transporter activity / potassium:proton antiporter activity / neurotransmitter transmembrane transporter activity / phosphate ion homeostasis / neural retina development / regulation of synapse structure or activity / monoatomic anion transport / chloride channel activity / chloride channel complex / excitatory synapse / synaptic transmission, glutamatergic / hippocampus development / synaptic vesicle membrane / synaptic vesicle / presynapse / early endosome / neuron projection / plasma membrane
Similarity search - Function
Major facilitator superfamily / Major Facilitator Superfamily / Major facilitator superfamily domain / Major facilitator superfamily (MFS) profile. / MFS transporter superfamily
Similarity search - Domain/homology
Vesicular glutamate transporter 2
Similarity search - Component
Biological speciesRattus norvegicus (Norway rat)
Methodsingle particle reconstruction / cryo EM / Resolution: 3.3 Å
AuthorsLi F / Stroud RM
Funding support United States, 2 items
OrganizationGrant numberCountry
National Institutes of Health/National Institute of Neurological Disorders and Stroke (NIH/NINDS)R01NS089713 United States
National Institutes of Health/National Institute of Mental Health (NIH/NIMH)K99MH119591 United States
CitationJournal: To Be Published
Title: Substrate Recognition, Directional Flux, and Allostery in a Synaptic Vesicle Glutamate Transporter
Authors: Li F / Stroud RM
History
DepositionDec 8, 2021-
Header (metadata) releaseDec 14, 2022-
Map releaseDec 14, 2022-
UpdateJun 5, 2024-
Current statusJun 5, 2024Processing site: RCSB / Status: Released

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Structure visualization

Supplemental images

emd_25666.png

Downloads & links

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Map

FileDownload / File: emd_25666.map.gz / Format: CCP4 / Size: 244.1 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Annotationsharpened map for model building
Voxel sizeX=Y=Z: 0.86 Å
Density
Contour LevelBy AUTHOR: 6.1
Minimum - Maximum-39.436999999999998 - 55.372149999999998
Average (Standard dev.)0.000000000000121 (±1.0)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderZYX
Origin000
Dimensions400400400
Spacing400400400
CellA=B=C: 344.0 Å
α=β=γ: 90.0 °

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Supplemental data

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Sample components

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Entire : complex of WT VGLUT2 with a Fab

EntireName: complex of WT VGLUT2 with a Fab
Components
  • Complex: complex of WT VGLUT2 with a Fab
    • Protein or peptide: Vesicular glutamate transporter 2

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Supramolecule #1: complex of WT VGLUT2 with a Fab

SupramoleculeName: complex of WT VGLUT2 with a Fab / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all
Source (natural)Organism: Rattus norvegicus (Norway rat)

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Macromolecule #1: Vesicular glutamate transporter 2

MacromoleculeName: Vesicular glutamate transporter 2 / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Rattus norvegicus (Norway rat)
Molecular weightTheoretical: 64.63727 KDa
Recombinant expressionOrganism: Spodoptera frugiperda (fall armyworm)
SequenceString: MESVKQRILA PGKEGIKNFA GKSLGQIYRV LEKKQDNRET IELTEDGKPL EVPEKKAPLC DCTCFGLPRR YIIAIMSGLG FCISFGIRC NLGVAIVDMV NNSTIHRGGK VIKEKAKFNW DPETVGMIHG SFFWGYIITQ IPGGYIASRL AANRVFGAAI L LTSTLNML ...String:
MESVKQRILA PGKEGIKNFA GKSLGQIYRV LEKKQDNRET IELTEDGKPL EVPEKKAPLC DCTCFGLPRR YIIAIMSGLG FCISFGIRC NLGVAIVDMV NNSTIHRGGK VIKEKAKFNW DPETVGMIHG SFFWGYIITQ IPGGYIASRL AANRVFGAAI L LTSTLNML IPSAARVHYG CVIFVRILQG LVEGVTYPAC HGIWSKWAPP LERSRLATTS FCGSYAGAVI AMPLAGILVQ YT GWSSVFY VYGSFGMVWY MFWLLVSYES PAKHPTITDE ERRYIEESIG ESANLLGAME KFKTPWRKFF TSMPVYAIIV ANF CRSWTF YLLLISQPAY FEEVFGFEIS KVGMLSAVPH LVMTIIVPIG GQIADFLRSK QILSTTTVRK IMNCGGFGME ATLL LVVGY SHTRGVAISF LVLAVGFSGF AISGFNVNHL DIAPRYASIL MGISNGVGTL SGMVCPIIVG AMTKNKSREE WQYVF LIAA LVHYGGVIFY ALFASGEKQP WADPEETSEE KCGFIHEDEL DEETGDITQN YINYGTTKSY GATSQENGGW PNGWEK KEE FVQESAQDAY SYKDRDDYS

UniProtKB: Vesicular glutamate transporter 2

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Experimental details

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Structure determination

Methodcryo EM
Processingsingle particle reconstruction
Aggregation stateparticle

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Sample preparation

Concentration0.5 mg/mL
BufferpH: 7.4
GridModel: Quantifoil R1.2/1.3 / Material: COPPER / Mesh: 400 / Pretreatment - Type: GLOW DISCHARGE
VitrificationCryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 283 K

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Image recordingFilm or detector model: GATAN K3 (6k x 4k) / Average electron dose: 60.0 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: DARK FIELD / Nominal defocus max: 20.0 µm / Nominal defocus min: 5.0 µm
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Startup modelType of model: PDB ENTRY
PDB model - PDB ID:

6v4d
PDB Unreleased entry

Final reconstructionResolution.type: BY AUTHOR / Resolution: 3.3 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 152461
Initial angle assignmentType: MAXIMUM LIKELIHOOD
Final angle assignmentType: MAXIMUM LIKELIHOOD
FSC plot (resolution estimation)

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Atomic model buiding 1

Initial modelPDB ID:

6v4d
PDB Unreleased entry


Chain - Source name: PDB / Chain - Initial model type: experimental model
Output model

PDB-7t3o:
Rat vesicular glutamate transporter 2 (VGLUT2) in low Cl condition

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