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Yorodumi- PDB-8vby: Structure of the human dopamine transporter in complex with beta-... -
+Open data
-Basic information
Entry | Database: PDB / ID: 8vby | |||||||||
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Title | Structure of the human dopamine transporter in complex with beta-CFT, MRS7292 and divalent zinc | |||||||||
Components | Sodium-dependent dopamine transporter | |||||||||
Keywords | MEMBRANE PROTEIN / Transport / Membrane / Inhibitor | |||||||||
Function / homology | Function and homology information Defective SLC6A3 causes Parkinsonism-dystonia infantile (PKDYS) / Defective SLC6A3 causes Parkinsonism-dystonia infantile (PKDYS) / Dopamine clearance from the synaptic cleft / amine binding / adenohypophysis development / dopamine uptake / hyaloid vascular plexus regression / dopamine binding / norepinephrine:sodium symporter activity / dopamine:sodium symporter activity ...Defective SLC6A3 causes Parkinsonism-dystonia infantile (PKDYS) / Defective SLC6A3 causes Parkinsonism-dystonia infantile (PKDYS) / Dopamine clearance from the synaptic cleft / amine binding / adenohypophysis development / dopamine uptake / hyaloid vascular plexus regression / dopamine binding / norepinephrine:sodium symporter activity / dopamine:sodium symporter activity / norepinephrine transport / regulation of dopamine metabolic process / neurotransmitter transmembrane transporter activity / dopamine transport / dopaminergic synapse / flotillin complex / dopamine catabolic process / monoamine transmembrane transporter activity / monoamine transport / Na+/Cl- dependent neurotransmitter transporters / positive regulation of multicellular organism growth / response to iron ion / neurotransmitter transport / dopamine biosynthetic process / amino acid transport / heterocyclic compound binding / dopamine uptake involved in synaptic transmission / neuronal cell body membrane / sodium ion transmembrane transport / prepulse inhibition / axon terminus / response to cAMP / lactation / protein phosphatase 2A binding / response to cocaine / cognition / locomotory behavior / response to nicotine / sensory perception of smell / presynaptic membrane / protease binding / postsynaptic membrane / response to ethanol / neuron projection / response to xenobiotic stimulus / membrane raft / axon / signaling receptor binding / neuronal cell body / protein-containing complex binding / cell surface / membrane / metal ion binding / plasma membrane / cytoplasm Similarity search - Function | |||||||||
Biological species | Homo sapiens (human) | |||||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.19 Å | |||||||||
Authors | Srivastava, D.K. / Gouaux, E. | |||||||||
Funding support | United States, 2items
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Citation | Journal: Nature / Year: 2024 Title: Structure of the human dopamine transporter and mechanisms of inhibition. Authors: Dushyant Kumar Srivastava / Vikas Navratna / Dilip K Tosh / Audrey Chinn / Md Fulbabu Sk / Emad Tajkhorshid / Kenneth A Jacobson / Eric Gouaux / Abstract: The neurotransmitter dopamine has central roles in mood, appetite, arousal and movement. Despite its importance in brain physiology and function, and as a target for illicit and therapeutic drugs, ...The neurotransmitter dopamine has central roles in mood, appetite, arousal and movement. Despite its importance in brain physiology and function, and as a target for illicit and therapeutic drugs, the human dopamine transporter (hDAT) and mechanisms by which it is inhibited by small molecules and Zn are without a high-resolution structural context. Here we determine the structure of hDAT in a tripartite complex with the competitive inhibitor and cocaine analogue, (-)-2-β-carbomethoxy-3-β-(4-fluorophenyl)tropane (β-CFT), the non-competitive inhibitor MRS7292 and Zn (ref. ). We show how β-CFT occupies the central site, approximately halfway across the membrane, stabilizing the transporter in an outward-open conformation. MRS7292 binds to a structurally uncharacterized allosteric site, adjacent to the extracellular vestibule, sequestered underneath the extracellular loop 4 (EL4) and adjacent to transmembrane helix 1b (TM1b), acting as a wedge, precluding movement of TM1b and closure of the extracellular gate. A Zn ion further stabilizes the outward-facing conformation by coupling EL4 to EL2, TM7 and TM8, thus providing specific insights into how Zn restrains the movement of EL4 relative to EL2 and inhibits transport activity. | |||||||||
History |
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-Structure visualization
Structure viewer | Molecule: MolmilJmol/JSmol |
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-Downloads & links
-Download
PDBx/mmCIF format | 8vby.cif.gz | 168.1 KB | Display | PDBx/mmCIF format |
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PDB format | pdb8vby.ent.gz | Display | PDB format | |
PDBx/mmJSON format | 8vby.json.gz | Tree view | PDBx/mmJSON format | |
Others | Other downloads |
-Validation report
Summary document | 8vby_validation.pdf.gz | 1.7 MB | Display | wwPDB validaton report |
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Full document | 8vby_full_validation.pdf.gz | 1.7 MB | Display | |
Data in XML | 8vby_validation.xml.gz | 38.6 KB | Display | |
Data in CIF | 8vby_validation.cif.gz | 54.3 KB | Display | |
Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/vb/8vby ftp://data.pdbj.org/pub/pdb/validation_reports/vb/8vby | HTTPS FTP |
-Related structure data
Related structure data | 43128MC M: map data used to model this data C: citing same article (ref.) |
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Similar structure data | Similarity search - Function & homologyF&H Search |
-Links
-Assembly
Deposited unit |
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1 |
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-Components
-Protein / Sugars , 2 types, 2 molecules A
#1: Protein | Mass: 63384.500 Da / Num. of mol.: 1 / Mutation: I248Y Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: SLC6A3 / Production host: Homo sapiens (human) / References: UniProt: Q01959 |
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#4: Sugar | ChemComp-NAG / |
-Non-polymers , 12 types, 50 molecules
#2: Chemical | ChemComp-42L / | ||||||||||||||||||
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#3: Chemical | ChemComp-A1AAF / Mass: 459.906 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: C20H18ClN5O4S / Feature type: SUBJECT OF INVESTIGATION | ||||||||||||||||||
#5: Chemical | ChemComp-Y01 / #6: Chemical | ChemComp-D10 / #7: Chemical | ChemComp-LNK / #8: Chemical | ChemComp-HP6 / #9: Chemical | ChemComp-ZN / | #10: Chemical | ChemComp-NA / | #11: Chemical | ChemComp-D12 / #12: Chemical | ChemComp-MYS / | #13: Chemical | #14: Water | ChemComp-HOH / | |
-Details
Has ligand of interest | Y |
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-Experimental details
-Experiment
Experiment | Method: ELECTRON MICROSCOPY |
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EM experiment | Aggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction |
-Sample preparation
Component | Name: Sodium dependent dopamine transporter in complex with inhibitor Type: COMPLEX / Entity ID: #1 / Source: RECOMBINANT | ||||||||||||||||||||
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Molecular weight | Value: 0.063 MDa / Experimental value: NO | ||||||||||||||||||||
Source (natural) | Organism: Homo sapiens (human) | ||||||||||||||||||||
Source (recombinant) | Organism: Homo sapiens (human) | ||||||||||||||||||||
Buffer solution | pH: 7.8 | ||||||||||||||||||||
Buffer component |
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Specimen | Conc.: 7 mg/ml / Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES / Details: monodisperse sample | ||||||||||||||||||||
Specimen support | Grid material: GOLD / Grid mesh size: 200 divisions/in. / Grid type: Quantifoil R1.2/1.3 | ||||||||||||||||||||
Vitrification | Instrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE / Humidity: 100 % / Chamber temperature: 288.15 K |
-Electron microscopy imaging
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
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Microscopy | Model: TFS KRIOS |
Electron gun | Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM |
Electron lens | Mode: BRIGHT FIELD / Nominal magnification: 165000 X / Nominal defocus max: 2500 nm / Nominal defocus min: 1000 nm / Cs: 2.7 mm |
Image recording | Electron dose: 50 e/Å2 / Film or detector model: FEI FALCON IV (4k x 4k) |
EM imaging optics | Energyfilter name: TFS Selectris X / Energyfilter slit width: 6 eV |
-Processing
EM software | Name: cryoSPARC / Category: CTF correction | ||||||||||||||||||||||||
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Image processing | Details: Falcon 4i | ||||||||||||||||||||||||
CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||
Symmetry | Point symmetry: C1 (asymmetric) | ||||||||||||||||||||||||
3D reconstruction | Resolution: 3.19 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 177494 / Symmetry type: POINT | ||||||||||||||||||||||||
Refinement | Cross valid method: NONE Stereochemistry target values: GeoStd + Monomer Library + CDL v1.2 | ||||||||||||||||||||||||
Displacement parameters | Biso mean: 53.23 Å2 | ||||||||||||||||||||||||
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