+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-20509 | |||||||||
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Title | MscS DDM | |||||||||
Map data | MscS Nanodisc with N-terminal His-Tag | |||||||||
Sample |
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Keywords | MscS / DDM / Mechanosensitive Channel of Small Conductance / Mechanosensitive / Channel / MEMBRANE PROTEIN | |||||||||
Function / homology | Function and homology information intracellular water homeostasis / mechanosensitive monoatomic ion channel activity / protein homooligomerization / monoatomic ion transmembrane transport / identical protein binding / membrane / plasma membrane Similarity search - Function | |||||||||
Biological species | Escherichia coli (E. coli) / Escherichia coli (strain K12) (bacteria) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.4 Å | |||||||||
Authors | Reddy BG / Perozo E | |||||||||
Funding support | United States, 1 items
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Citation | Journal: Elife / Year: 2019 Title: Molecular basis of force-from-lipids gating in the mechanosensitive channel MscS. Authors: Bharat Reddy / Navid Bavi / Allen Lu / Yeonwoo Park / Eduardo Perozo / Abstract: Prokaryotic mechanosensitive (MS) channels open by sensing the physical state of the membrane. As such, lipid-protein interactions represent the defining molecular process underlying ...Prokaryotic mechanosensitive (MS) channels open by sensing the physical state of the membrane. As such, lipid-protein interactions represent the defining molecular process underlying mechanotransduction. Here, we describe cryo-electron microscopy (cryo-EM) structures of the small-conductance mechanosensitive channel (MscS) in nanodiscs (ND). They reveal a novel membrane-anchoring fold that plays a significant role in channel activation and establish a new location for the lipid bilayer, shifted ~14 Å from previous consensus placements. Two types of lipid densities are explicitly observed. A phospholipid that 'hooks' the top of each TM2-TM3 hairpin and likely plays a role in force sensing, and a bundle of acyl chains occluding the permeation path above the L105 cuff. These observations reshape our understanding of force-from-lipids gating in MscS and highlight the key role of allosteric interactions between TM segments and phospholipids bound to key dynamic components of the channel. | |||||||||
History |
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-Structure visualization
Movie |
Movie viewer |
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Structure viewer | EM map: SurfViewMolmilJmol/JSmol |
Supplemental images |
-Downloads & links
-EMDB archive
Map data | emd_20509.map.gz | 49.1 MB | EMDB map data format | |
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Header (meta data) | emd-20509-v30.xml emd-20509.xml | 15 KB 15 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_20509_fsc.xml | 9.1 KB | Display | FSC data file |
Images | emd_20509.png | 144.4 KB | ||
Filedesc metadata | emd-20509.cif.gz | 5.4 KB | ||
Others | emd_20509_half_map_1.map.gz emd_20509_half_map_2.map.gz | 49.6 MB 49.6 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-20509 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-20509 | HTTPS FTP |
-Validation report
Summary document | emd_20509_validation.pdf.gz | 820.6 KB | Display | EMDB validaton report |
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Full document | emd_20509_full_validation.pdf.gz | 820.2 KB | Display | |
Data in XML | emd_20509_validation.xml.gz | 16.2 KB | Display | |
Data in CIF | emd_20509_validation.cif.gz | 21.1 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-20509 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-20509 | HTTPS FTP |
-Related structure data
Related structure data | 6pwoMC 6pwnC 6pwpC C: citing same article (ref.) M: atomic model generated by this map |
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Similar structure data |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_20509.map.gz / Format: CCP4 / Size: 64 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | MscS Nanodisc with N-terminal His-Tag | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.064 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Half map: Unfiltered Half Map 1
File | emd_20509_half_map_1.map | ||||||||||||
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Annotation | Unfiltered Half Map 1 | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Unfiltered Half Map 2
File | emd_20509_half_map_2.map | ||||||||||||
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Annotation | Unfiltered Half Map 2 | ||||||||||||
Projections & Slices |
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Density Histograms |
-Sample components
-Entire : Mechanosensitive Channel of Small Conductance
Entire | Name: Mechanosensitive Channel of Small Conductance |
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Components |
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-Supramolecule #1: Mechanosensitive Channel of Small Conductance
Supramolecule | Name: Mechanosensitive Channel of Small Conductance / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
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Source (natural) | Organism: Escherichia coli (E. coli) |
-Macromolecule #1: Small-conductance mechanosensitive channel
Macromolecule | Name: Small-conductance mechanosensitive channel / type: protein_or_peptide / ID: 1 / Number of copies: 7 / Enantiomer: LEVO |
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Source (natural) | Organism: Escherichia coli (strain K12) (bacteria) / Strain: K12 |
Molecular weight | Theoretical: 31.205178 KDa |
Recombinant expression | Organism: Escherichia coli (E. coli) |
Sequence | String: GSHMEDLNVV DSINGAGSWL VANQALLLSY AVNIVAALAI IIVGLIIARM ISNAVNRLMI SRKIDATVAD FLSALVRYGI IAFTLIAAL GRVGVQTASV IAVLGAAGLA VGLALQGSLS NLAAGVLLVM FRPFRAGEYV DLGGVAGTVL SVQIFSTTMR T ADGKIIVI ...String: GSHMEDLNVV DSINGAGSWL VANQALLLSY AVNIVAALAI IIVGLIIARM ISNAVNRLMI SRKIDATVAD FLSALVRYGI IAFTLIAAL GRVGVQTASV IAVLGAAGLA VGLALQGSLS NLAAGVLLVM FRPFRAGEYV DLGGVAGTVL SVQIFSTTMR T ADGKIIVI PNGKIIAGNI INFSREPVRR NEFIIGVAYD SDIDQVKQIL TNIIQSEDRI LKDREMTVRL NELGASSINF VV RVWSNSG DLQNVYWDVL ERIKREFDAA GISFPYPQMD VNFKRVKEDK AA UniProtKB: Small-conductance mechanosensitive channel |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Concentration | 2 mg/mL |
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Buffer | pH: 7.4 |
Grid | Model: Quantifoil R2/2 / Material: COPPER / Mesh: 200 / Pretreatment - Type: GLOW DISCHARGE |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 295 K / Instrument: FEI VITROBOT MARK IV Details: 22C. Blot Force 3 for 3 seconds. Double application with a blotting between applications.. |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Average electron dose: 50.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
+Image processing
-Atomic model buiding 1
Refinement | Space: REAL / Protocol: FLEXIBLE FIT |
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Output model | PDB-6pwo: |