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- EMDB-29885: CLC-ec1 R230C/L249C/C85A at pH 4.5 100mM Cl Turn -

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Basic information

Entry
Database: EMDB / ID: EMD-29885
TitleCLC-ec1 R230C/L249C/C85A at pH 4.5 100mM Cl Turn
Map data
Sample
  • Cell: Structure of ecCLC-R230C/L249C/C85A mutant at pH 4.5 in 100mM Cl
    • Protein or peptide: H(+)/Cl(-) exchange transporter ClcA
  • Ligand: CHLORIDE ION
KeywordsCLC-ec1 / ecCLC / eriC / CLC transporter / chloride proton antiporter / TRANSPORT PROTEIN
Function / homologyChloride channel, ClcA / Chloride channel, voltage gated / Chloride channel, core / Voltage gated chloride channel / voltage-gated chloride channel activity / antiporter activity / plasma membrane / H(+)/Cl(-) exchange transporter ClcA
Function and homology information
Biological speciesEscherichia coli (E. coli)
Methodsingle particle reconstruction / cryo EM / Resolution: 3.1 Å
AuthorsFortea E / Lee S / Argyos Y / Chadda R / Ciftci D / Huysmans G / Robertson JL / Boudker O / Accardi A
Funding support United States, 1 items
OrganizationGrant numberCountry
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)R01 GM128420 United States
CitationJournal: Nat Struct Mol Biol / Year: 2024
Title: Structural basis of pH-dependent activation in a CLC transporter.
Authors: Eva Fortea / Sangyun Lee / Rahul Chadda / Yiorgos Argyros / Priyanka Sandal / Robyn Mahoney-Kruszka / Hatice Didar Ciftci / Maria E Falzone / Gerard Huysmans / Janice L Robertson / Olga ...Authors: Eva Fortea / Sangyun Lee / Rahul Chadda / Yiorgos Argyros / Priyanka Sandal / Robyn Mahoney-Kruszka / Hatice Didar Ciftci / Maria E Falzone / Gerard Huysmans / Janice L Robertson / Olga Boudker / Alessio Accardi /
Abstract: CLCs are dimeric chloride channels and anion/proton exchangers that regulate processes such as muscle contraction and endo-lysosome acidification. Common gating controls their activity; its closure ...CLCs are dimeric chloride channels and anion/proton exchangers that regulate processes such as muscle contraction and endo-lysosome acidification. Common gating controls their activity; its closure simultaneously silences both protomers, and its opening allows them to independently transport ions. Mutations affecting common gating in human CLCs cause dominant genetic disorders. The structural rearrangements underlying common gating are unknown. Here, using single-particle cryo-electron microscopy, we show that the prototypical Escherichia coli CLC-ec1 undergoes large-scale rearrangements in activating conditions. The slow, pH-dependent remodeling of the dimer interface leads to the concerted opening of the intracellular H pathways and is required for transport. The more frequent formation of short water wires in the open H pathway enables Cl pore openings. Mutations at disease-causing sites favor CLC-ec1 activation and accelerate common gate opening in the human CLC-7 exchanger. We suggest that the pH activation mechanism of CLC-ec1 is related to the common gating of CLC-7.
History
DepositionFeb 22, 2023-
Header (metadata) releaseFeb 7, 2024-
Map releaseFeb 7, 2024-
UpdateOct 16, 2024-
Current statusOct 16, 2024Processing site: RCSB / Status: Released

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Structure visualization

Supplemental images

emd_29885.png

Downloads & links

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Map

FileDownload / File: emd_29885.map.gz / Format: CCP4 / Size: 64 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
0.83 Å/pix.
x 256 pix.
= 211.2 Å		0.83 Å/pix.
x 256 pix.
= 211.2 Å		0.83 Å/pix.
x 256 pix.
= 211.2 Å

Surface

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Images are generated by Spider.

Voxel sizeX=Y=Z: 0.825 Å
Density

Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 0.4
Minimum - Maximum-2.6387243 - 3.708055
Average (Standard dev.)0.004840137 (±0.09022804)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions256256256
Spacing256256256
CellA=B=C: 211.2 Å
α=β=γ: 90.0 °

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Supplemental data

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Half map: #1

Fileemd_29885_half_map_1.map
Projections & Slices
AxesZYX

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Density Histograms

Histogram

Histogram (log scale)

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Half map: #2

Fileemd_29885_half_map_2.map
Projections & Slices
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Sample components

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Entire : Structure of ecCLC-R230C/L249C/C85A mutant at pH 4.5 in 100mM Cl

EntireName: Structure of ecCLC-R230C/L249C/C85A mutant at pH 4.5 in 100mM Cl
Components
  • Cell: Structure of ecCLC-R230C/L249C/C85A mutant at pH 4.5 in 100mM Cl
    • Protein or peptide: H(+)/Cl(-) exchange transporter ClcA
  • Ligand: CHLORIDE ION

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Supramolecule #1: Structure of ecCLC-R230C/L249C/C85A mutant at pH 4.5 in 100mM Cl

SupramoleculeName: Structure of ecCLC-R230C/L249C/C85A mutant at pH 4.5 in 100mM Cl
type: cell / ID: 1 / Parent: 0 / Macromolecule list: #1
Source (natural)Organism: Escherichia coli (E. coli)

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Macromolecule #1: H(+)/Cl(-) exchange transporter ClcA

MacromoleculeName: H(+)/Cl(-) exchange transporter ClcA / type: protein_or_peptide / ID: 1 / Number of copies: 2 / Enantiomer: LEVO
Source (natural)Organism: Escherichia coli (E. coli)
Molecular weightTheoretical: 49.118027 KDa
Recombinant expressionOrganism: Escherichia coli (E. coli)
SequenceString: MKTDTPSLET PQAARLRRRQ LIRQLLERDK TPLAILFMAA VVGTLVGLAA VAFDKGVAWL QNQRMGALVH TADNYPLLLT VAFLASAVL AMFGYFLVRK YAPEAGGSGI PEIEGALEDQ RPVRWWRVLP VKFFGGLGTL GGGMVLGREG PTVQIGGNIG R MVLDIFRL ...String:
MKTDTPSLET PQAARLRRRQ LIRQLLERDK TPLAILFMAA VVGTLVGLAA VAFDKGVAWL QNQRMGALVH TADNYPLLLT VAFLASAVL AMFGYFLVRK YAPEAGGSGI PEIEGALEDQ RPVRWWRVLP VKFFGGLGTL GGGMVLGREG PTVQIGGNIG R MVLDIFRL KGDEARHTLL ATGAAAGLAA AFNAPLAGIL FIIEEMRPQF RYTLISIKAV FIGVIMSTIM YCIFNHEVAL ID VGKLSDA PCNTLWLYLI LGIIFGIFGP IFNKWVLGMQ DLLHRVHGGN ITKWVLMGGA IGGLCGLLGF VAPATSGGGF NLI PIATAG NFSMGMLVFI FVARVITTLL CFSSGAPGGI FAPMLALGTV LGTAFGMVAV ELFPQYHLEA GTFAIAGMGA LLAA SIRAP LTGIILVLEM TDNYQLILPM IITGLGATLL AQFTGGKPLY SAILARTLAK QEAEQL

UniProtKB: H(+)/Cl(-) exchange transporter ClcA

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Macromolecule #2: CHLORIDE ION

MacromoleculeName: CHLORIDE ION / type: ligand / ID: 2 / Number of copies: 4 / Formula: CL
Molecular weightTheoretical: 35.453 Da

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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

BufferpH: 4.5
VitrificationCryogen name: ETHANE

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Image recordingFilm or detector model: GATAN K3 (6k x 4k) / Average electron dose: 56.11 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 2.0 µm / Nominal defocus min: 0.7000000000000001 µm
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Startup modelType of model: NONE
Final reconstructionApplied symmetry - Point group: C1 (asymmetric) / Resolution.type: BY AUTHOR / Resolution: 3.1 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 235275
Initial angle assignmentType: MAXIMUM LIKELIHOOD
Final angle assignmentType: MAXIMUM LIKELIHOOD

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