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- EMDB-63170: Cryo-EM Structure of GAT3 -

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Basic information

Entry
Database: EMDB / ID: EMD-63170
TitleCryo-EM Structure of GAT3
Map data
Sample
  • Complex: structure of GAT3
    • Protein or peptide: Sodium- and chloride-dependent GABA transporter 3
Keywordsprotein structure / STRUCTURAL PROTEIN / TRANSPORT PROTEIN
Function / homology
Function and homology information


gamma-aminobutyric acid reuptake / Reuptake of GABA / monocarboxylic acid transmembrane transporter activity / monocarboxylic acid transport / Creatine metabolism / taurine:sodium symporter activity / gamma-aminobutyric acid:sodium:chloride symporter activity / SLC-mediated transport of neurotransmitters / amino acid binding / amino acid transport ...gamma-aminobutyric acid reuptake / Reuptake of GABA / monocarboxylic acid transmembrane transporter activity / monocarboxylic acid transport / Creatine metabolism / taurine:sodium symporter activity / gamma-aminobutyric acid:sodium:chloride symporter activity / SLC-mediated transport of neurotransmitters / amino acid binding / amino acid transport / cell projection / sodium ion transmembrane transport / GABA-ergic synapse / presynaptic membrane / postsynaptic membrane / response to xenobiotic stimulus / membrane / plasma membrane
Similarity search - Function
Sodium:neurotransmitter symporter, GABA, GAT-3 / Sodium:neurotransmitter symporter family signature 2. / Sodium:neurotransmitter symporter family signature 1. / Sodium:neurotransmitter symporter / Sodium:neurotransmitter symporter superfamily / Sodium:neurotransmitter symporter family / Sodium:neurotransmitter symporter family profile.
Similarity search - Domain/homology
Sodium- and chloride-dependent GABA transporter 3
Similarity search - Component
Biological speciesHomo sapiens (human)
Methodsingle particle reconstruction / cryo EM / Resolution: 3.6 Å
AuthorsZhao Y / Xu H
Funding support China, 1 items
OrganizationGrant numberCountry
National Natural Science Foundation of China (NSFC) China
CitationJournal: Structure / Year: 2025
Title: Substrate and inhibitor binding of human GABA transporter 3.
Authors: Hao Xu / Yimin Zhang / Qinru Bai / Lingli He / Qihao Chen / Yunlong Qiu / Renjie Li / Jie Yu / Jun Zhao / Yan Zhao /
Abstract: GABA (g-aminobutyric acid) transporter 3 (GAT3) is primarily found in glial cells and is essential for regulating GABA homeostasis in the central nervous system by mediating GABA uptake. ...GABA (g-aminobutyric acid) transporter 3 (GAT3) is primarily found in glial cells and is essential for regulating GABA homeostasis in the central nervous system by mediating GABA uptake. Consequently, GAT3 has emerged as a significant therapeutic target for the treatment of epilepsy. In this study, we present the cryoelectron microscopy (cryo-EM) structures of GAT3 bound to its substrate GABA, the selective inhibitor SNAP-5114, and in the substrate-free state. GAT3 binds to GABA in an inward-facing conformation, while SNAP-5114 occupies the GABA-binding pocket and is stabilized by extensive interactions with surrounding residues. Functional studies reveal that E66 plays a pivotal role in determining the substrate-binding mode and specificity of SNAP-5114 binding. Taken together, our study clarifies the GABA binding mechanism of GAT3 and reveals the molecular basis for the specific inhibition of SNAP-5114, offering valuable insights for developing GAT3 subtypes selective inhibitors, which hold potential as a treatment for epilepsy.
History
DepositionJan 16, 2025-
Header (metadata) releaseAug 20, 2025-
Map releaseAug 20, 2025-
UpdateMar 4, 2026-
Current statusMar 4, 2026Processing site: PDBj / Status: Released

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Structure visualization

Supplemental images

emd_63170.png

Downloads & links

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Map

FileDownload / File: emd_63170.map.gz / Format: CCP4 / Size: 64 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Projections & slices

Image control

Size
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AxesZ (Sec.)Y (Row.)X (Col.)
0.85 Å/pix.
x 256 pix.
= 217.6 Å		0.85 Å/pix.
x 256 pix.
= 217.6 Å		0.85 Å/pix.
x 256 pix.
= 217.6 Å

Surface

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Images are generated by Spider.

Voxel sizeX=Y=Z: 0.85 Å
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Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 0.134
Minimum - Maximum-0.86161494 - 1.2624896
Average (Standard dev.)0.00080783037 (±0.034576945)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions256256256
Spacing256256256
CellA=B=C: 217.6 Å
α=β=γ: 90.0 °

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Supplemental data

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Half map: #2

Fileemd_63170_half_map_1.map
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Half map: #1

Fileemd_63170_half_map_2.map
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Sample components

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Entire : structure of GAT3

EntireName: structure of GAT3
Components
  • Complex: structure of GAT3
    • Protein or peptide: Sodium- and chloride-dependent GABA transporter 3

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Supramolecule #1: structure of GAT3

SupramoleculeName: structure of GAT3 / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all
Source (natural)Organism: Homo sapiens (human)

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Macromolecule #1: Sodium- and chloride-dependent GABA transporter 3

MacromoleculeName: Sodium- and chloride-dependent GABA transporter 3 / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 70.628312 KDa
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString: MTAEKALPLG NGKAAEEARE SEAPGGGSSS GGAAPARHPR VKRDKAVHER GHWNNKVEFV LSVAGEIIGL GNVWRFPYLC YKNGGGAFL IPYVVFFICC GIPVFFLETA LGQFTSEGGI TCWRKVCPLF EGIGYATQVI EAHLNVYYII ILAWAIFYLS N CFTTELPW ...String:
MTAEKALPLG NGKAAEEARE SEAPGGGSSS GGAAPARHPR VKRDKAVHER GHWNNKVEFV LSVAGEIIGL GNVWRFPYLC YKNGGGAFL IPYVVFFICC GIPVFFLETA LGQFTSEGGI TCWRKVCPLF EGIGYATQVI EAHLNVYYII ILAWAIFYLS N CFTTELPW ATCGHEWNTE NCVEFQKLNV SNYSHVSLQN ATSPVMEFWE HRVLAISDGI EHIGNLRWEL ALCLLAAWTI CY FCIWKGT KSTGKVVYVT ATFPYIMLLI LLIRGVTLPG ASEGIKFYLY PDLSRLSDPQ VWVDAGTQIF FSYAICLGCL TAL GSYNNY NNNCYRDCIM LCCLNSGTSF VAGFAIFSVL GFMAYEQGVP IAEVAESGPG LAFIAYPKAV TMMPLSPLWA TLFF MMLIF LGLDSQFVCV ESLVTAVVDM YPKVFRRGYR RELLILALSV ISYFLGLVML TEGGMYIFQL FDSYAASGMC LLFVA IFEC ICIGWVYGSN RFYDNIEDMI GYRPPSLIKW CWMIMTPGIC AGIFIFFLIK YKPLKYNNIY TYPAWGYGIG WLMALS SML CIPLWICITV WKTEGTLPEK LQKLTTPSTD LKMRGKLGVS PRMVTVNDSD AKLKSDGTIA AITEKETHF

UniProtKB: Sodium- and chloride-dependent GABA transporter 3

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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

BufferpH: 8
VitrificationCryogen name: ETHANE

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Electron microscopy

MicroscopeFEI TALOS ARCTICA
Image recordingFilm or detector model: GATAN K3 BIOCONTINUUM (6k x 4k) / Average electron dose: 60.0 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: OTHER / Imaging mode: BRIGHT FIELD / Nominal defocus max: 2.0 µm / Nominal defocus min: 1.0 µm
Experimental equipment
Model: Talos Arctica / Image courtesy: FEI Company

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Image processing

CTF correctionType: PHASE FLIPPING ONLY
Startup modelType of model: OTHER
Final reconstructionResolution.type: BY AUTHOR / Resolution: 3.6 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: cryoSPARC (ver. 4) / Number images used: 35082
Initial angle assignmentType: MAXIMUM LIKELIHOOD
Final angle assignmentType: MAXIMUM LIKELIHOOD

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