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Open data
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Basic information
| Entry | ![]() | ||||||||||||
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| Title | pentameric MscL from Escherichia coli in nanodiscs | ||||||||||||
Map data | sharpened map in phenix: -50 | ||||||||||||
Sample |
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Keywords | mechanosensitive channel / bacteria / osmotic shock / MEMBRANE PROTEIN | ||||||||||||
| Function / homology | Function and homology informationintracellular water homeostasis / mechanosensitive monoatomic ion channel activity / monoatomic ion transport / monoatomic ion transmembrane transport / membrane / identical protein binding / plasma membrane Similarity search - Function | ||||||||||||
| Biological species | ![]() | ||||||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 3.12 Å | ||||||||||||
Authors | Rasmussen T / Flegler VJ / Bottcher B | ||||||||||||
| Funding support | Germany, 3 items
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Citation | Journal: J Mol Biol / Year: 2026Title: The Structure of Escherichia coli MscL and Its Dimer Formation in Nanodiscs. Authors: Tim Rasmussen / Julia Isabel Bahner / Vanessa J Flegler / Tamsanqa T Hove / Christian Kraft / Akiko Rasmussen / Bettina Böttcher / ![]() Abstract: Mechanosensitive channels of large conductance (MscL) are essential bacterial safety valves that prevent osmotic lysis by releasing solutes in response to membrane tension. Despite extensive ...Mechanosensitive channels of large conductance (MscL) are essential bacterial safety valves that prevent osmotic lysis by releasing solutes in response to membrane tension. Despite extensive functional studies on Escherichia coli MscL (EcMscL), its high-resolution structure remained unknown. Using cryo-electron microscopy, we present an experimental structure of EcMscL reconstituted in nanodiscs at 3.1 Å resolution. The structure reveals a pentameric assembly with a narrow hydrophobic gate at the cytosolic side and a periplasmic cavity, consistent with the canonical MscL-fold. Differences to earlier published crystal structures of MscL from other organisms are in the less conserved periplasmic loop. We observe a previously unreported dimeric association of EcMscL pentamers, mediated by residues 61-63 in the periplasmic loop. This dimeric interface is located at the periplasmic side and provides a structural basis for the formation of higher-order clusters. The observed arrangement enables a fluid-like, mosaic packing of channels with centre-to-centre distances of 5.9-9 nm, consistent with biophysical and imaging data. These findings provide a structural framework for understanding cluster organisation of EcMscL that modulates its activity in cellular stress response. | ||||||||||||
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Structure visualization
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Downloads & links
-EMDB archive
| Map data | emd_56958.map.gz | 54 MB | EMDB map data format | |
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| Header (meta data) | emd-56958-v30.xml emd-56958.xml | 20.2 KB 20.2 KB | Display Display | EMDB header |
| FSC (resolution estimation) | emd_56958_fsc.xml | 8.4 KB | Display | FSC data file |
| Images | emd_56958.png | 72.5 KB | ||
| Filedesc metadata | emd-56958.cif.gz | 6 KB | ||
| Others | emd_56958_half_map_1.map.gz emd_56958_half_map_2.map.gz | 59.3 MB 59.3 MB | ||
| Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-56958 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-56958 | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 28ycMC ![]() 56936 M: atomic model generated by this map C: citing same article ( |
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| Similar structure data | Similarity search - Function & homology F&H Search |
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Links
| EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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| Related items in Molecule of the Month |
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Map
| File | Download / File: emd_56958.map.gz / Format: CCP4 / Size: 64 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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| Annotation | sharpened map in phenix: -50 | ||||||||||||||||||||||||||||||||||||
| Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
| Voxel size | X=Y=Z: 0.946 Å | ||||||||||||||||||||||||||||||||||||
| Density |
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| Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
| Details | EMDB XML:
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-Supplemental data
-Half map: half map A
| File | emd_56958_half_map_1.map | ||||||||||||
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| Annotation | half map A | ||||||||||||
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-Half map: half map B
| File | emd_56958_half_map_2.map | ||||||||||||
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| Annotation | half map B | ||||||||||||
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| Density Histograms |
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Sample components
-Entire : pentameric MscL in nanodiscs MSP1E3D1
| Entire | Name: pentameric MscL in nanodiscs MSP1E3D1 |
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| Components |
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-Supramolecule #1: pentameric MscL in nanodiscs MSP1E3D1
| Supramolecule | Name: pentameric MscL in nanodiscs MSP1E3D1 / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
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| Source (natural) | Organism: ![]() |
| Molecular weight | Theoretical: 75 KDa |
-Macromolecule #1: Large-conductance mechanosensitive channel
| Macromolecule | Name: Large-conductance mechanosensitive channel / type: protein_or_peptide / ID: 1 / Details: C-terminal His6-tag / Number of copies: 5 / Enantiomer: LEVO |
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| Source (natural) | Organism: ![]() |
| Molecular weight | Theoretical: 15.801593 KDa |
| Recombinant expression | Organism: ![]() |
| Sequence | String: MSIIKEFREF AMRGNVVDLA VGVIIGAAFG KIVSSLVADI IMPPLGLLIG GIDFKQFAVT LRDAQGDIPA VVMHYGVFIQ NVFDFLIVA FAIFMAIKLI NKLNRKKEEP AAAPAPTKEE VLLTEIRDLL KEQNNRSHHH HHH UniProtKB: Large-conductance mechanosensitive channel |
-Experimental details
-Structure determination
| Method | cryo EM |
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Processing | single particle reconstruction |
| Aggregation state | particle |
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Sample preparation
| Concentration | 1 mg/mL | |||||||||
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| Buffer | pH: 7.5 Component:
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| Grid | Model: Quantifoil R1.2/1.3 / Material: GOLD / Mesh: 300 / Support film - Material: GOLD / Support film - topology: HOLEY ARRAY / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 150 sec. / Pretreatment - Atmosphere: AIR / Pretreatment - Pressure: 0.4 kPa | |||||||||
| Vitrification | Cryogen name: ETHANE / Chamber humidity: 90 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK IV / Details: 5 sec at blot force +20. |
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Electron microscopy
| Microscope | TFS KRIOS |
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| Specialist optics | Energy filter - Name: TFS Selectris / Energy filter - Slit width: 5 eV |
| Image recording | Film or detector model: FEI FALCON IV (4k x 4k) / Number grids imaged: 1 / Number real images: 14428 / Average exposure time: 5.89 sec. / Average electron dose: 70.0 e/Å2 |
| Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
| Electron optics | C2 aperture diameter: 70.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 1.6 µm / Nominal defocus min: 0.5 µm / Nominal magnification: 130000 |
| Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
| Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
-Atomic model buiding 1
| Initial model | Chain - Source name: Other / Chain - Initial model type: other / Details: Modelangelo |
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| Refinement | Space: REAL / Protocol: AB INITIO MODEL / Overall B value: 123 / Target criteria: Cross-correlation |
| Output model | ![]() PDB-28yc: |
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Keywords
Authors
Germany, 3 items
Citation



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Y (Row.)
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FIELD EMISSION GUN

