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- EMDB-10703: Structure of Human Potassium Chloride Transporter KCC3 S45D/T940D... -

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Basic information

Entry
Database: EMDB / ID: EMD-10703
TitleStructure of Human Potassium Chloride Transporter KCC3 S45D/T940D/T997D in NaCl
Map data
Sample
  • Complex: Homodimeric complex of human potassium chloride transporter
    • Protein or peptide: Solute carrier family 12 member 6
  • Ligand: 2-acetamido-2-deoxy-beta-D-glucopyranose
Function / homology
Function and homology information


Defective SLC12A6 causes agenesis of the corpus callosum, with peripheral neuropathy (ACCPN) / ammonium import across plasma membrane / potassium ion transmembrane transporter activity / potassium:chloride symporter activity / Cation-coupled Chloride cotransporters / chloride ion homeostasis / cellular hypotonic response / ammonium transmembrane transporter activity / cellular hypotonic salinity response / potassium ion homeostasis ...Defective SLC12A6 causes agenesis of the corpus callosum, with peripheral neuropathy (ACCPN) / ammonium import across plasma membrane / potassium ion transmembrane transporter activity / potassium:chloride symporter activity / Cation-coupled Chloride cotransporters / chloride ion homeostasis / cellular hypotonic response / ammonium transmembrane transporter activity / cellular hypotonic salinity response / potassium ion homeostasis / cell volume homeostasis / potassium ion import across plasma membrane / monoatomic ion transport / potassium ion transmembrane transport / chloride transmembrane transport / cellular response to glucose stimulus / chemical synaptic transmission / basolateral plasma membrane / angiogenesis / axon / synapse / protein kinase binding / membrane / metal ion binding / plasma membrane
Similarity search - Function
K/Cl co-transporter / SLC12A transporter family / SLC12A transporter, C-terminal / Solute carrier family 12 / Amino acid permease/ SLC12A domain / Amino acid permease
Similarity search - Domain/homology
Solute carrier family 12 member 6
Similarity search - Component
Biological speciesHomo sapiens (human)
Methodsingle particle reconstruction / cryo EM / Resolution: 3.76 Å
AuthorsChi G / Man H / Ebenhoch R / Reggiano G / Pike ACW / Wang D / McKinley G / Mukhopadhyay SMM / MacLean B / Chalk R ...Chi G / Man H / Ebenhoch R / Reggiano G / Pike ACW / Wang D / McKinley G / Mukhopadhyay SMM / MacLean B / Chalk R / Moreau C / Snee M / Bohstedt T / Singh NK / Abrusci P / Arrowsmith CH / Bountra C / Edwards AM / Marsden BD / Burgess-Brown NA / DiMaio F / Duerr KL / Structural Genomics Consortium (SGC)
Funding support United Kingdom, 1 items
OrganizationGrant numberCountry
European Commission115766 United Kingdom
CitationJournal: To be published
Title: Structure of Human Potassium Chloride Transporter KCC3 in NaCl
Authors: Chi G / Man H / Ebenhoch R / Reggiano G / Pike ACW / Wang D / McKinley G / Mukhopadhyay SMM / Chalk R / Moreau C / Snee M / Bohstedt T / Singh NK / Abrusci P / Arrowsmith CH / Bountra C / ...Authors: Chi G / Man H / Ebenhoch R / Reggiano G / Pike ACW / Wang D / McKinley G / Mukhopadhyay SMM / Chalk R / Moreau C / Snee M / Bohstedt T / Singh NK / Abrusci P / Arrowsmith CH / Bountra C / Edwards AM / Marsden BD / Burgess-Brown NA / DiMaio F / Duerr KL / Structural Genomics Consortium (SGC)
History
DepositionFeb 25, 2020-
Header (metadata) releaseMar 11, 2020-
Map releaseMar 11, 2020-
UpdateDec 2, 2020-
Current statusDec 2, 2020Processing site: PDBe / Status: Released

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Structure visualization

Movie
  • Surface view with section colored by density value
  • Surface level: 0.01
  • Imaged by UCSF Chimera
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  • Surface view colored by cylindrical radius
  • Surface level: 0.01
  • Imaged by UCSF Chimera
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  • Surface view with fitted model
  • Atomic models: PDB-6y5r
  • Surface level: 0.01
  • Imaged by UCSF Chimera
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Movie viewer
Structure viewerEM map:
SurfViewMolmilJmol/JSmol
Supplemental images

emd_10703.png

Downloads & links

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Map

FileDownload / File: emd_10703.map.gz / Format: CCP4 / Size: 325 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Voxel sizeX=Y=Z: 0.651 Å
Density
Contour LevelBy AUTHOR: 0.01 / Movie #1: 0.01
Minimum - Maximum-0.020708172 - 0.042289127
Average (Standard dev.)0.0000602144 (±0.0012882311)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions440440440
Spacing440440440
CellA=B=C: 286.44 Å
α=β=γ: 90.0 °

CCP4 map header:

modeImage stored as Reals
Å/pix. X/Y/Z0.6510.6510.651
M x/y/z440440440
origin x/y/z0.0000.0000.000
length x/y/z286.440286.440286.440
α/β/γ90.00090.00090.000
start NX/NY/NZ-200-200-200
NX/NY/NZ401401401
MAP C/R/S123
start NC/NR/NS000
NC/NR/NS440440440
D min/max/mean-0.0210.0420.000

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Supplemental data

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Half map: #1

Fileemd_10703_half_map_1.map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Half map: #2

Fileemd_10703_half_map_2.map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Sample components

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Entire : Homodimeric complex of human potassium chloride transporter

EntireName: Homodimeric complex of human potassium chloride transporter
Components
  • Complex: Homodimeric complex of human potassium chloride transporter
    • Protein or peptide: Solute carrier family 12 member 6
  • Ligand: 2-acetamido-2-deoxy-beta-D-glucopyranose

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Supramolecule #1: Homodimeric complex of human potassium chloride transporter

SupramoleculeName: Homodimeric complex of human potassium chloride transporter
type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1
Source (natural)Organism: Homo sapiens (human)
Recombinant expressionOrganism: Homo sapiens (human)
Molecular weightTheoretical: 246 KDa

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Macromolecule #1: Solute carrier family 12 member 6

MacromoleculeName: Solute carrier family 12 member 6 / type: protein_or_peptide / ID: 1
Details: Sugar molecules from glycosylation post-translational modifications (NAG, BMA) present.
Number of copies: 2 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 122.273344 KDa
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString: MPHFTVTKVE DPEEGAAASI SQEPSLADIK ARIQDSDEPD LSQNDITGEH SQLLDDGHKK ARNAYLNNSN YEEGDEYFDK NLALFEEEM DTRPKVSSLL NRMANYTNLT QGAKEHEEAE NITEGKKKPT KTPQMGTFMG VYLPCLQNIF GVILFLRLTW V VGTAGVLQ ...String:
MPHFTVTKVE DPEEGAAASI SQEPSLADIK ARIQDSDEPD LSQNDITGEH SQLLDDGHKK ARNAYLNNSN YEEGDEYFDK NLALFEEEM DTRPKVSSLL NRMANYTNLT QGAKEHEEAE NITEGKKKPT KTPQMGTFMG VYLPCLQNIF GVILFLRLTW V VGTAGVLQ AFAIVLICCC CTMLTAISMS AIATNGVVPA GGSYFMISRA LGPEFGGAVG LCFYLGTTFA AAMYILGAIE IF LVYIVPR AAIFHSDDAL KESAAMLNNM RVYGTAFLVL MVLVVFIGVR YVNKFASLFL ACVIVSILAI YAGAIKSSFA PPH FPVCML GNRTLSSRHI DVCSKTKEIN NMTVPSKLWG FFCNSSQFFN ATCDEYFVHN NVTSIQGIPG LASGIITENL WSNY LPKGE IIEKPSAKSS DVLGSLNHEY VLVDITTSFT LLVGIFFPSV TGIMAGSNRS GDLKDAQKSI PIGTILAILT TSFVY LSNV VLFGACIEGV VLRDKFGDAV KGNLVVGTLS WPSPWVIVIG SFFSTCGAGL QSLTGAPRLL QAIAKDNIIP FLRVFG HSK ANGEPTWALL LTAAIAELGI LIASLDLVAP ILSMFFLMCY LFVNLACALQ TLLRTPNWRP RFRYYHWALS FMGMSIC LA LMFISSWYYA IVAMVIAGMI YKYIEYQGAE KEWGDGIRGL SLSAARFALL RLEEGPPHTK NWRPQLLVLL KLDEDLHV K HPRLLTFASQ LKAGKGLTIV GSVIVGNFLE NYGEALAAEQ TIKHLMEAEK VKGFCQLVVA AKLREGISHL IQSCGLGGM KHNTVVMGWP NGWRQSEDAR AWKTFIGTVR VTTAAHLALL VAKNISFFPS NVEQFSEGNI DVWWIVHDGG MLMLLPFLLK QHKVWRKCS IRIFTVAQLE DNSIQMKKDL ATFLYHLRIE AEVEVVEMHD SDISAYTYER DLMMEQRSQM LRHMRLSKTE R DREAQLVK DRNSMLRLTS IGSDEDEETE TYQEKVHMDW TKDKYMASRG QKAKSMEGFQ DLLNMRPDQS NVRRMHTAVK LN EVIVNKS HEAKLVLLNM PGPPRNPEGD ENYMEFLEVL TEGLERVLLV RGGGSEVITI YS

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Macromolecule #3: 2-acetamido-2-deoxy-beta-D-glucopyranose

MacromoleculeName: 2-acetamido-2-deoxy-beta-D-glucopyranose / type: ligand / ID: 3 / Number of copies: 2 / Formula: NAG
Molecular weightTheoretical: 221.208 Da
Chemical component information

ChemComp-NAG:
2-acetamido-2-deoxy-beta-D-glucopyranose / N-Acetylglucosamine

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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

Concentration6.5 mg/mL
BufferpH: 7.5
GridModel: Quantifoil R1.2/1.3 / Material: GOLD / Mesh: 300 / Support film - Material: CARBON / Support film - topology: HOLEY / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Atmosphere: OTHER
VitrificationCryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 297 K / Instrument: FEI VITROBOT MARK IV
Details: 2 second blotting time, 40 second waiting time, -15 blotting force.
DetailsThis sample was monodisperse.

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Electron microscopy

MicroscopeTFS KRIOS
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsC2 aperture diameter: 50.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELDBright-field microscopy / Cs: 2.7 mm / Nominal defocus max: -2.3 µm / Nominal defocus min: -0.8 µm
Sample stageSpecimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN
TemperatureMin: 70.0 K / Max: 70.0 K
Image recordingFilm or detector model: GATAN K3 (6k x 4k) / Number grids imaged: 1 / Number real images: 16472 / Average electron dose: 40.0 e/Å2
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

CTF correctionSoftware - Name: Gctf (ver. 1.16)
Startup modelType of model: NONE
Details: map was built ab initio using Relion 3.0.8's initial model program
Initial angle assignmentType: RANDOM ASSIGNMENT / Software - Name: RELION (ver. 3.0.8)
Final 3D classificationSoftware - Name: RELION (ver. 3.0.8)
Final angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: RELION (ver. 3.0.8)
Final reconstructionApplied symmetry - Point group: C1 (asymmetric) / Resolution.type: BY AUTHOR / Resolution: 3.76 Å / Resolution method: FSC 0.5 CUT-OFF / Software - Name: RELION (ver. 3.0.8) / Number images used: 920574
FSC plot (resolution estimation)

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