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- PDB-7d14: Mouse KCC2 -

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Basic information

Entry
Database: PDB / ID: 7d14
TitleMouse KCC2
ComponentsSolute carrier family 12 member 5
KeywordsMEMBRANE PROTEIN / transporter
Function / homology
Function and homology information


Cation-coupled Chloride cotransporters / thermosensory behavior / intracellular pH reduction / potassium:chloride symporter activity / hypotonic response / intracellular chloride ion homeostasis / ammonium transmembrane transport / ammonium channel activity / chloride ion homeostasis / dendritic spine development ...Cation-coupled Chloride cotransporters / thermosensory behavior / intracellular pH reduction / potassium:chloride symporter activity / hypotonic response / intracellular chloride ion homeostasis / ammonium transmembrane transport / ammonium channel activity / chloride ion homeostasis / dendritic spine development / potassium ion homeostasis / cell volume homeostasis / postsynaptic specialization membrane / chloride transport / potassium ion import across plasma membrane / monoatomic ion transport / chloride transmembrane transport / dendrite membrane / potassium ion transmembrane transport / learning / cell periphery / potassium ion transport / multicellular organism growth / chemical synaptic transmission / perikaryon / response to xenobiotic stimulus / neuronal cell body / membrane / metal ion binding / plasma membrane
Similarity search - Function
K/Cl co-transporter / SLC12A transporter, C-terminal / Solute carrier family 12 / Amino acid permease/ SLC12A domain / SLC12A transporter family / Amino acid permease
Similarity search - Domain/homology
Solute carrier family 12 member 5
Similarity search - Component
Biological speciesMus musculus (house mouse)
MethodELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.8 Å
AuthorsZhang, S. / Yang, M.
CitationJournal: Commun Biol / Year: 2021
Title: The structural basis of function and regulation of neuronal cotransporters NKCC1 and KCC2.
Authors: Sensen Zhang / Jun Zhou / Yuebin Zhang / Tianya Liu / Perrine Friedel / Wei Zhuo / Suma Somasekharan / Kasturi Roy / Laixing Zhang / Yang Liu / Xianbin Meng / Haiteng Deng / Wenwen Zeng / ...Authors: Sensen Zhang / Jun Zhou / Yuebin Zhang / Tianya Liu / Perrine Friedel / Wei Zhuo / Suma Somasekharan / Kasturi Roy / Laixing Zhang / Yang Liu / Xianbin Meng / Haiteng Deng / Wenwen Zeng / Guohui Li / Biff Forbush / Maojun Yang /
Abstract: NKCC and KCC transporters mediate coupled transport of Na+K+Cl and K+Cl across the plasma membrane, thus regulating cell Cl concentration and cell volume and playing critical roles in transepithelial ...NKCC and KCC transporters mediate coupled transport of Na+K+Cl and K+Cl across the plasma membrane, thus regulating cell Cl concentration and cell volume and playing critical roles in transepithelial salt and water transport and in neuronal excitability. The function of these transporters has been intensively studied, but a mechanistic understanding has awaited structural studies of the transporters. Here, we present the cryo-electron microscopy (cryo-EM) structures of the two neuronal cation-chloride cotransporters human NKCC1 (SLC12A2) and mouse KCC2 (SLC12A5), along with computational analysis and functional characterization. These structures highlight essential residues in ion transport and allow us to propose mechanisms by which phosphorylation regulates transport activity.
History
DepositionSep 13, 2020Deposition site: PDBJ / Processing site: PDBJ
Revision 1.0Apr 14, 2021Provider: repository / Type: Initial release
Revision 1.1May 29, 2024Group: Data collection / Database references / Category: chem_comp_atom / chem_comp_bond / database_2
Item: _database_2.pdbx_DOI / _database_2.pdbx_database_accession

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Structure visualization

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Assembly

Deposited unit
A: Solute carrier family 12 member 5
B: Solute carrier family 12 member 5


Theoretical massNumber of molelcules
Total (without water)252,8052
Polymers252,8052
Non-polymers00
Water00
1


  • Idetical with deposited unit
  • defined by author
  • Evidence: gel filtration
TypeNameSymmetry operationNumber
identity operation1_5551

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Components

#1: Protein Solute carrier family 12 member 5 / Electroneutral potassium-chloride cotransporter 2 / K-Cl cotransporter 2 / mKCC2 / Neuronal K-Cl cotransporter


Mass: 126402.328 Da / Num. of mol.: 2
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Mus musculus (house mouse) / Gene: Slc12a5, Kcc2, Kiaa1176 / Production host: Homo sapiens (human) / References: UniProt: Q91V14

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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: mouse KCC2 / Type: ORGANELLE OR CELLULAR COMPONENT / Entity ID: all / Source: RECOMBINANT
Source (natural)Organism: Mus musculus (house mouse)
Source (recombinant)Organism: Homo sapiens (human)
Buffer solutionpH: 7.4
SpecimenEmbedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES
VitrificationCryogen name: ETHANE

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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: BRIGHT FIELD
Image recordingElectron dose: 50 e/Å2 / Film or detector model: GATAN K2 SUMMIT (4k x 4k)

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Processing

CTF correctionType: PHASE FLIPPING AND AMPLITUDE CORRECTION
3D reconstructionResolution: 3.8 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 66000 / Symmetry type: POINT

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