+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-18046 | |||||||||
---|---|---|---|---|---|---|---|---|---|---|
Title | Wait Complex: BAM bound Darobactin-B and Extended SurA | |||||||||
Map data | ||||||||||
Sample |
| |||||||||
Keywords | Outer Membrane / Complex / Chaperone / Protein Folding / Inhibitor / MEMBRANE PROTEIN | |||||||||
Function / homology | Function and homology information Bam protein complex / Gram-negative-bacterium-type cell outer membrane assembly / protein insertion into membrane / peptidylprolyl isomerase / peptidyl-prolyl cis-trans isomerase activity / cell outer membrane / peptide binding / unfolded protein binding / protein folding / outer membrane-bounded periplasmic space ...Bam protein complex / Gram-negative-bacterium-type cell outer membrane assembly / protein insertion into membrane / peptidylprolyl isomerase / peptidyl-prolyl cis-trans isomerase activity / cell outer membrane / peptide binding / unfolded protein binding / protein folding / outer membrane-bounded periplasmic space / protein-macromolecule adaptor activity / protein stabilization / response to antibiotic / cell surface / identical protein binding / membrane Similarity search - Function | |||||||||
Biological species | Escherichia coli (E. coli) / Photorhabdus heterorhabditis (bacteria) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 2.9 Å | |||||||||
Authors | Fenn KL / Ranson NA | |||||||||
Funding support | United Kingdom, 1 items
| |||||||||
Citation | Journal: Nat Commun / Year: 2024 Title: Outer membrane protein assembly mediated by BAM-SurA complexes. Authors: Katherine L Fenn / Jim E Horne / Joel A Crossley / Nils Böhringer / Romany J Horne / Till F Schäberle / Antonio N Calabrese / Sheena E Radford / Neil A Ranson / Abstract: The outer membrane is a formidable barrier that protects Gram-negative bacteria against environmental threats. Its integrity requires the correct folding and insertion of outer membrane proteins ...The outer membrane is a formidable barrier that protects Gram-negative bacteria against environmental threats. Its integrity requires the correct folding and insertion of outer membrane proteins (OMPs) by the membrane-embedded β-barrel assembly machinery (BAM). Unfolded OMPs are delivered to BAM by the periplasmic chaperone SurA, but how SurA and BAM work together to ensure successful OMP delivery and folding remains unclear. Here, guided by AlphaFold2 models, we use disulphide bond engineering in an attempt to trap SurA in the act of OMP delivery to BAM, and solve cryoEM structures of a series of complexes. The results suggest that SurA binds BAM at its soluble POTRA-1 domain, which may trigger conformational changes in both BAM and SurA that enable transfer of the unfolded OMP to the BAM lateral gate for insertion into the outer membrane. Mutations that disrupt the interaction between BAM and SurA result in outer membrane assembly defects, supporting the key role of SurA in outer membrane biogenesis. | |||||||||
History |
|
-Structure visualization
Supplemental images |
---|
-Downloads & links
-EMDB archive
Map data | emd_18046.map.gz | 7.8 MB | EMDB map data format | |
---|---|---|---|---|
Header (meta data) | emd-18046-v30.xml emd-18046.xml | 26.6 KB 26.6 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_18046_fsc.xml | 10.7 KB | Display | FSC data file |
Images | emd_18046.png | 137.2 KB | ||
Filedesc metadata | emd-18046.cif.gz | 9.6 KB | ||
Others | emd_18046_half_map_1.map.gz emd_18046_half_map_2.map.gz | 80.8 MB 80.8 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-18046 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-18046 | HTTPS FTP |
-Validation report
Summary document | emd_18046_validation.pdf.gz | 788.6 KB | Display | EMDB validaton report |
---|---|---|---|---|
Full document | emd_18046_full_validation.pdf.gz | 788.1 KB | Display | |
Data in XML | emd_18046_validation.xml.gz | 17.8 KB | Display | |
Data in CIF | emd_18046_validation.cif.gz | 23.5 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-18046 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-18046 | HTTPS FTP |
-Related structure data
Related structure data | 8pzvMC 8pz1C 8pz2C 8pzuC 8q0gC 8qp5C 8qpuC 8qpvC 8qpwC C: citing same article (ref.) M: atomic model generated by this map |
---|---|
Similar structure data | Similarity search - Function & homologyF&H Search |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
---|---|
Related items in Molecule of the Month |
-Map
File | Download / File: emd_18046.map.gz / Format: CCP4 / Size: 103 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 0.91 Å | ||||||||||||||||||||||||||||||||||||
Density |
| ||||||||||||||||||||||||||||||||||||
Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
|
-Supplemental data
-Half map: #2
File | emd_18046_half_map_1.map | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Projections & Slices |
| ||||||||||||
Density Histograms |
-Half map: #1
File | emd_18046_half_map_2.map | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Projections & Slices |
| ||||||||||||
Density Histograms |
-Sample components
+Entire : Wait Complex: BAM bound Darobactin-B and Extended SurA
+Supramolecule #1: Wait Complex: BAM bound Darobactin-B and Extended SurA
+Macromolecule #1: Outer membrane protein assembly factor BamA
+Macromolecule #2: Outer membrane protein assembly factor BamB
+Macromolecule #3: Outer membrane protein assembly factor BamC
+Macromolecule #4: Outer membrane protein assembly factor BamD
+Macromolecule #5: Outer membrane protein assembly factor BamE
+Macromolecule #6: Chaperone SurA
+Macromolecule #7: Darobactin-B
+Macromolecule #8: MAGNESIUM ION
+Macromolecule #9: water
-Experimental details
-Structure determination
Method | cryo EM |
---|---|
Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Concentration | 3 mg/mL |
---|---|
Buffer | pH: 8 |
Grid | Model: Quantifoil R1.2/1.3 / Material: COPPER / Mesh: 300 / Pretreatment - Type: PLASMA CLEANING / Pretreatment - Time: 60 sec. |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 90 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK IV |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
---|---|
Specialist optics | Energy filter - Name: TFS Selectris / Energy filter - Slit width: 10 eV |
Software | Name: EPU |
Image recording | Film or detector model: FEI FALCON IV (4k x 4k) / Number grids imaged: 1 / Number real images: 6357 / Average electron dose: 39.8 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 3.0 µm / Nominal defocus min: 0.9 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 |
+Image processing
-Atomic model buiding 1
Initial model |
| ||||||||
---|---|---|---|---|---|---|---|---|---|
Software | Name: UCSF ChimeraX | ||||||||
Refinement | Space: REAL / Protocol: RIGID BODY FIT / Overall B value: 82 | ||||||||
Output model | PDB-8pzv: |