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- PDB-9cax: Structure of human SLC2A9 transporter -

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Basic information

Entry
Database: PDB / ID: 9cax
TitleStructure of human SLC2A9 transporter
ComponentsSoluble cytochrome b562,Solute carrier family 2, facilitated glucose transporter member 9
KeywordsMEMBRANE PROTEIN / Transporter
Function / homology
Function and homology information


Defective SLC2A9 causes hypouricemia renal 2 (RHUC2) / fructose transmembrane transporter activity / fructose transmembrane transport / hexose transmembrane transport / dehydroascorbic acid transport / carbohydrate:proton symporter activity / Cellular hexose transport / D-glucose transmembrane transporter activity / D-glucose transmembrane transport / urate transport ...Defective SLC2A9 causes hypouricemia renal 2 (RHUC2) / fructose transmembrane transporter activity / fructose transmembrane transport / hexose transmembrane transport / dehydroascorbic acid transport / carbohydrate:proton symporter activity / Cellular hexose transport / D-glucose transmembrane transporter activity / D-glucose transmembrane transport / urate transport / urate metabolic process / urate transmembrane transporter activity / D-glucose import / transmembrane transporter activity / electron transport chain / basolateral plasma membrane / periplasmic space / electron transfer activity / apical plasma membrane / iron ion binding / heme binding / membrane / plasma membrane
Similarity search - Function
Glucose transporter GLUT / Sugar/inositol transporter / Sugar transport proteins signature 2. / Sugar transport proteins signature 1. / Sugar transporter, conserved site / Major facilitator, sugar transporter-like / Sugar (and other) transporter / Major facilitator superfamily domain / Major facilitator superfamily (MFS) profile. / Cytochrome b562 ...Glucose transporter GLUT / Sugar/inositol transporter / Sugar transport proteins signature 2. / Sugar transport proteins signature 1. / Sugar transporter, conserved site / Major facilitator, sugar transporter-like / Sugar (and other) transporter / Major facilitator superfamily domain / Major facilitator superfamily (MFS) profile. / Cytochrome b562 / Cytochrome b562 / MFS transporter superfamily / Cytochrome c/b562
Similarity search - Domain/homology
Soluble cytochrome b562 / Solute carrier family 2, facilitated glucose transporter member 9
Similarity search - Component
Biological speciesHomo sapiens (human)
MethodELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.37 Å
AuthorsKhandelwal, N.K. / Gupta, M. / Stroud, R.M.
Funding support United States, 1items
OrganizationGrant numberCountry
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS) United States
CitationJournal: To Be Published
Title: Structure of human SLC2A9
Authors: Khandelwal, N.K. / Stroud, R.M. / Gupta, M.G.
History
DepositionJun 18, 2024Deposition site: RCSB / Processing site: RCSB
Revision 1.0Jan 29, 2025Provider: repository / Type: Initial release
Revision 1.0Jan 29, 2025Data content type: EM metadata / Data content type: EM metadata / Provider: repository / Type: Initial release
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Revision 1.0Jan 29, 2025Data content type: FSC / Data content type: FSC / Provider: repository / Type: Initial release
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Revision 1.0Jan 29, 2025Data content type: Image / Data content type: Image / Provider: repository / Type: Initial release
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Revision 1.1May 28, 2025Group: Data collection / Category: em_admin / em_software / Item: _em_admin.last_update / _em_software.name
Revision 1.1May 28, 2025Data content type: EM metadata / Data content type: EM metadata / EM metadata / Group: Data processing / Experimental summary / Data content type: EM metadata / EM metadata / Category: em_admin / em_software / Data content type: EM metadata / EM metadata / Item: _em_admin.last_update / _em_software.name

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: Soluble cytochrome b562,Solute carrier family 2, facilitated glucose transporter member 9


Theoretical massNumber of molelcules
Total (without water)69,8601
Polymers69,8601
Non-polymers00
Water00
1


  • Idetical with deposited unit
  • defined by author&software
  • Evidence: electron microscopy, not applicable
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1

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Components

#1: Protein Soluble cytochrome b562,Solute carrier family 2, facilitated glucose transporter member 9 / Cytochrome b-562 / Glucose transporter type 9 / GLUT-9 / Urate transporter


Mass: 69859.695 Da / Num. of mol.: 1
Source method: isolated from a genetically manipulated source
Details: N-terminal Flag tag, HRV3C site followed by BRIL tag N-terminal truncated SlC2A9 isoform 1 (residue 52-540 as per uniprot). C terminal thrombin site followed by 10X his tag,N-terminal Flag ...Details: N-terminal Flag tag, HRV3C site followed by BRIL tag N-terminal truncated SlC2A9 isoform 1 (residue 52-540 as per uniprot). C terminal thrombin site followed by 10X his tag,N-terminal Flag tag, HRV3C site followed by BRIL tag N-terminal truncated SlC2A9 isoform 1 (residue 52-540 as per uniprot). C terminal thrombin site followed by 10X his tag,N-terminal Flag tag, HRV3C site followed by BRIL tag N-terminal truncated SlC2A9 isoform 1 (residue 52-540 as per uniprot). C terminal thrombin site followed by 10X his tag,N-terminal Flag tag, HRV3C site followed by BRIL tag N-terminal truncated SlC2A9 isoform 1 (residue 52-540 as per uniprot). C terminal thrombin site followed by 10X his tag,N-terminal Flag tag, HRV3C site followed by BRIL tag N-terminal truncated SlC2A9 isoform 1 (residue 52-540 as per uniprot). C terminal thrombin site followed by 10X his tag,N-terminal Flag tag, HRV3C site followed by BRIL tag N-terminal truncated SlC2A9 isoform 1 (residue 52-540 as per uniprot). C terminal thrombin site followed by 10X his tag,N-terminal Flag tag, HRV3C site followed by BRIL tag N-terminal truncated SlC2A9 isoform 1 (residue 52-540 as per uniprot). C terminal thrombin site followed by 10X his tag,N-terminal Flag tag, HRV3C site followed by BRIL tag N-terminal truncated SlC2A9 isoform 1 (residue 52-540 as per uniprot). C terminal thrombin site followed by 10X his tag
Source: (gene. exp.) Homo sapiens (human) / Gene: cybC, SLC2A9, GLUT9 / Plasmid: 83 nu vector / Production host: Saccharomyces cerevisiae (brewer's yeast) / References: UniProt: P0ABE7, UniProt: Q9NRM0
Has protein modificationN

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Experimental details

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Experiment

ExperimentMethod: ELECTRON MICROSCOPY
EM experimentAggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction

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Sample preparation

ComponentName: Codon optimize SLC2A9 gene was synthesize and cloned in 83 nu yeast expressing vector from GenScript. Protein expressed in yeast cells and purified using Ni-affinity followed by Sizing.
Type: COMPLEX / Entity ID: all / Source: RECOMBINANT
Molecular weightValue: 0.06978906 MDa / Experimental value: NO
Source (natural)Organism: Homo sapiens (Human) (human)
Source (recombinant)Organism: Saccharomyces cerevisiae (brewer's yeast)
Buffer solutionpH: 7.5
Details: 4 degree cold buffer pH7.5 (300mM NaCl, 50mM Tris and 0.02% GDN)
Buffer component
IDConc.NameFormulaBuffer-ID
1300 mMSodium ChlorideNaCl1
250 mMTrisTris-Cl1
SpecimenConc.: 7 mg/ml / Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES
Specimen supportDetails: 30 second glow 30 sec hold / Grid material: GOLD / Grid mesh size: 400 divisions/in. / Grid type: Quantifoil R1.2/1.3
VitrificationInstrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE / Humidity: 100 % / Chamber temperature: 277.15 K

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Electron microscopy imaging

Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company
MicroscopyModel: FEI TITAN KRIOS
Electron gunElectron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM
Electron lensMode: OTHER / Nominal defocus max: 2000 nm / Nominal defocus min: 1000 nm / Cs: 2.7 mm / C2 aperture diameter: 70 µm
Specimen holderCryogen: NITROGEN
Image recordingAverage exposure time: 2 sec. / Electron dose: 47.7 e/Å2 / Film or detector model: GATAN K3 (6k x 4k) / Num. of grids imaged: 1 / Num. of real images: 10503

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Processing

EM software
IDNameCategory
1cryoSPARCparticle selection
8PHENIXmodel refinement
CTF correctionType: NONE
Particle selectionNum. of particles selected: 9590974
3D reconstructionResolution: 3.37 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 154457 / Symmetry type: POINT
Refine LS restraints
Refine-IDTypeDev idealNumber
ELECTRON MICROSCOPYf_bond_d0.0043702
ELECTRON MICROSCOPYf_angle_d0.9225044
ELECTRON MICROSCOPYf_dihedral_angle_d5.059504
ELECTRON MICROSCOPYf_chiral_restr0.049595
ELECTRON MICROSCOPYf_plane_restr0.007625

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