NYU Dataset

Substrate Translocation and Inhibition in Human Dicarboxylate Transporter NaDC3

Part of: Wang Lab |
UID: 10727
* Corresponding Author
Description

High-affinity sodium–dicarboxylate cotransporter (NaDC3) imports various substrates into the cell as tricarboxylate acid cycle intermediates, lipid biosynthesis precursors, and signaling molecules. To understand the cellular signaling process and developing inhibitors requires knowledge of the structural basis of the dicarboxylate specificity and inhibition mechanism of NaDC3. This study used single-particle cryo-electron microscopy to determine structures of NaDC3 in the presence of each of the three substrates: succinate, DMS and αKG. The dataset includes electron microscopy densities and protein models of NaDC3–succinate Coo–Coo, NaDC3–succinate Coo–Ci, NaDC3–DMS Co–Ci, NaDC3–DMS Ci–Ci, NaDC3–αKG Ci–Ci, NaDC3–PF4a Ci–Ci, NaCT–succinate Ci–Ci, and NaCT–PF4a Ci–Ci complexes. This helped determine the structures of NaDC3 in various dimers, revealing the protomer in three conformations: outward-open Co, outward-occluded Coo, and inward-open Ci.

Subject of Study
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Instructions
Electron microscopy densities and protein models were deposited to the Electron Microscopy Data Bank (EMDB) and Protein Data Bank (PDB).
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NaDC3–succinate Coo–Coo complex
Accession #: EMD-42621

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NaDC3–succinate Coo–Ci complex
Accession #: EMD-42617

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NaDC3–DMS Co–Ci complex
Accession #: EMD-42619

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NaDC3–DMS Ci–Ci complex
Accession #: EMD-42618

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NaDC3–αKG Ci–Ci complex
Accession #: EMD-42615

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NaDC3–PF4a Ci–Ci complex
Accession #: EMD-42616

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NaCT–succinate Ci–Ci complex
Accession #: EMD-42620

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NaCT–PF4a Ci–Ci complex
Accession #: EMD-42614

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NaDC3–succinate Coo–Coo complex
Accession #: 8UVI

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NaDC3–succinate Coo–Ci complex
Accession #: 8UVE

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NaDC3–DMS Co–Ci complex
Accession #: 8UVG

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NaDC3–DMS Ci–Ci complex
Accession #: 8UVF

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NaDC3–αKG Ci–Ci complex
Accession #: 8UVC

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NaDC3–PF4a Ci–Ci complex
Accession #: 8UVD

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NaCT–succinate Ci–Ci complex
Accession #: 8UVH

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NaCT–PF4a Ci–Ci complex
Accession #: 8UVB

Associated Publications
Data Type
Equipment Used
Bruker Avance 800 MHz NMR
HORIBA FluoroMax-4
Nikon Eclipse Ti
Thermo Scientific Talos Arctica
Thermo Scientific Titan Krios
Thermo Scientific Vitrobot Mark IV
Software Used
AlphaFold2
Chemical Sketch v1.0
ChimeraX v1.4
Coot v0.9.6
CryoSPARC v3.3.1
CTFFIND4 v4.1.13
GraphPad Prism v9.4.1
ISOLDE v1.0
Leginon v3.6
MotionCor2 v1.5
Phenix v1.19
PyMOL v2.5.2
RELION v4.0
TopSpin v4.1.1
WARP v1.0.9
Grant Support
Simons Foundation/Simons Foundation
TESS Research Foundation/TESS Research Foundation
Intramural Research Program/National Institute of Neurological Disorders and Stroke
G. Harold and Leila Y. Mathers Foundation/G. Harold and Leila Y. Mathers Foundation