+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-15040 | |||||||||
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Title | BcsHD cis-filaments: four 'beads-on-a-string' | |||||||||
Map data | BcsHD 'beads-on-a-string' filaments : Unsharpened locally refined map of a BcsH-bound tetramer of BcsD octamers | |||||||||
Sample |
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Biological species | Komagataeibacter hansenii ATCC 23769 (bacteria) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 2.77 Å | |||||||||
Authors | Krasteva PV / Abidi W / Decossas M | |||||||||
Funding support | European Union, 1 items
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Citation | Journal: Sci Adv / Year: 2022 Title: Bacterial crystalline cellulose secretion via a supramolecular BcsHD scaffold. Authors: Wiem Abidi / Marion Decossas / Lucía Torres-Sánchez / Lucie Puygrenier / Sylvie Létoffé / Jean-Marc Ghigo / Petya V Krasteva / Abstract: Cellulose, the most abundant biopolymer on Earth, is not only the predominant constituent of plants but also a key extracellular polysaccharide in the biofilms of many bacterial species. Depending on ...Cellulose, the most abundant biopolymer on Earth, is not only the predominant constituent of plants but also a key extracellular polysaccharide in the biofilms of many bacterial species. Depending on the producers, chemical modifications, and three-dimensional assemblies, bacterial cellulose (BC) can present diverse degrees of crystallinity. Highly ordered, or crystalline, cellulose presents great economical relevance due to its ever-growing number of biotechnological applications. Even if some acetic acid bacteria have long been identified as BC superproducers, the molecular mechanisms determining the secretion of crystalline versus amorphous cellulose remain largely unknown. Here, we present structural and mechanistic insights into the role of the accessory subunits BcsH (CcpAx) and BcsD (CesD) that determine crystalline BC secretion in the lineage. We show that oligomeric BcsH drives the assembly of BcsD into a supramolecular cytoskeletal scaffold that likely stabilizes the cellulose-extruding synthase nanoarrays through an unexpected inside-out mechanism for secretion system assembly. | |||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_15040.map.gz | 275.2 MB | EMDB map data format | |
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Header (meta data) | emd-15040-v30.xml emd-15040.xml | 17.4 KB 17.4 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_15040_fsc.xml | 17.2 KB | Display | FSC data file |
Images | emd_15040.png | 48.9 KB | ||
Masks | emd_15040_msk_1.map | 548.9 MB | Mask map | |
Others | emd_15040_additional_1.map.gz emd_15040_half_map_1.map.gz emd_15040_half_map_2.map.gz | 518.8 MB 509.1 MB 509.1 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-15040 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-15040 | HTTPS FTP |
-Validation report
Summary document | emd_15040_validation.pdf.gz | 619.5 KB | Display | EMDB validaton report |
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Full document | emd_15040_full_validation.pdf.gz | 619.1 KB | Display | |
Data in XML | emd_15040_validation.xml.gz | 26.9 KB | Display | |
Data in CIF | emd_15040_validation.cif.gz | 35.5 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-15040 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-15040 | HTTPS FTP |
-Related structure data
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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-Map
File | Download / File: emd_15040.map.gz / Format: CCP4 / Size: 548.9 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||
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Annotation | BcsHD 'beads-on-a-string' filaments : Unsharpened locally refined map of a BcsH-bound tetramer of BcsD octamers | ||||||||||||||||||||
Voxel size | X=Y=Z: 1.05676 Å | ||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Mask #1
File | emd_15040_msk_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Additional map: BcsHD 'beads-on-a-string' filaments :Autosharpened locally refined map of...
File | emd_15040_additional_1.map | ||||||||||||
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Annotation | BcsHD 'beads-on-a-string' filaments :Autosharpened locally refined map of a BcsH-bound tetramer of BcsD octamers | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: BcsHD 'beads-on-a-string' filaments : half-map B
File | emd_15040_half_map_1.map | ||||||||||||
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Annotation | BcsHD 'beads-on-a-string' filaments : half-map B | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: BcsHD 'beads-on-a-string' filaments : half-map A
File | emd_15040_half_map_2.map | ||||||||||||
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Annotation | BcsHD 'beads-on-a-string' filaments : half-map A | ||||||||||||
Projections & Slices |
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Density Histograms |
-Sample components
-Entire : BcsHD 'beads-on-a-string' cis-filaments
Entire | Name: BcsHD 'beads-on-a-string' cis-filaments |
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Components |
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-Supramolecule #1: BcsHD 'beads-on-a-string' cis-filaments
Supramolecule | Name: BcsHD 'beads-on-a-string' cis-filaments / type: complex / ID: 1 / Chimera: Yes / Parent: 0 / Macromolecule list: all Details: BcsHD filaments purified upon coexpression of BcsD (no tag) and BcsH C-terminal region carrying an HRV3c-cleavable N-terminal hexahistidine tag. |
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Source (natural) | Organism: Komagataeibacter hansenii ATCC 23769 (bacteria) / Strain: ATCC 23769 / Location in cell: cytoskeleton |
-Macromolecule #1: BcsHD 'beads-on-a-string' cis-filaments
Macromolecule | Name: BcsHD 'beads-on-a-string' cis-filaments / type: other / ID: 1 Details: Multiple copies of BcsD and BcsH (protein in sample is tag-free with sequence: PMGSTKTDTNSSQASRPGSPVASPDGSPTMAEVFMTLGGRATELLSPRPSLREALLRRRENEEES) in BcsH-driven cis-filaments Classification: other |
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Source (natural) | Organism: Komagataeibacter hansenii ATCC 23769 (bacteria) / Strain: ATCC 23769 |
Sequence | String: BcsD MGSTK TDTNSSQASR PGSPVASPDG SPTMAEVFMT LGGRATELLS PRPSLREALL RRRENEEES BcsH MSYYH HHHHHDYDIP TTLEVLFQGP MGSTKTDTNS SQASRPGSPV ASPDGSPTMA EVFMTLGGRA TELLSPRPSL REALLRRREN EEES |
Recombinant expression | Organism: Escherichia coli BL21 (bacteria) |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | filament |
-Sample preparation
Concentration | 0.5 mg/mL |
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Buffer | pH: 8 / Details: 20 mM HEPES pH 8.0, 100 mM NaCl |
Grid | Model: Quantifoil R1.2/1.3 / Material: GOLD |
Vitrification | Cryogen name: ETHANE / Chamber temperature: 20 K / Instrument: FEI VITROBOT MARK IV |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Specialist optics | Energy filter - Name: GIF Quantum LS |
Image recording | Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Average electron dose: 49.9 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 2.75 µm / Nominal defocus min: 0.48 µm |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |