+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-11356 | ||||||||||||||||||
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Title | Structure of detergent-extracted full-length E.coli BcsB | ||||||||||||||||||
Map data | Main map non-uniformly refined. It represents two BcsB octamers (6YG8) linked by a central miscellaneous structure. | ||||||||||||||||||
Sample |
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Function / homology | Function and homology information cellulose biosynthetic process / UDP-alpha-D-glucose metabolic process / membrane => GO:0016020 / plasma membrane Similarity search - Function | ||||||||||||||||||
Biological species | Escherichia coli (E. coli) | ||||||||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 5.9 Å | ||||||||||||||||||
Authors | Abidi W / Zouhir S / Krasteva P | ||||||||||||||||||
Funding support | 5 items
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Citation | Journal: Sci Adv / Year: 2021 Title: Architecture and regulation of an enterobacterial cellulose secretion system. Authors: Wiem Abidi / Samira Zouhir / Meryem Caleechurn / Stéphane Roche / Petya Violinova Krasteva / Abstract: Many free-living and pathogenic enterobacteria secrete biofilm-promoting cellulose using a multicomponent, envelope-embedded Bcs secretion system under the control of intracellular second messenger c- ...Many free-living and pathogenic enterobacteria secrete biofilm-promoting cellulose using a multicomponent, envelope-embedded Bcs secretion system under the control of intracellular second messenger c-di-GMP. The molecular understanding of system assembly and cellulose secretion has been largely limited to the crystallographic studies of a distantly homologous BcsAB synthase tandem and a low-resolution reconstruction of an assembled macrocomplex that encompasses most of the inner membrane and cytosolic subunits and features an atypical layered architecture. Here, we present cryo-EM structures of the assembled Bcs macrocomplex, as well as multiple crystallographic snapshots of regulatory Bcs subcomplexes. The structural and functional data uncover the mechanism of asymmetric secretion system assembly and periplasmic crown polymerization and reveal unexpected subunit stoichiometry, multisite c-di-GMP recognition, and ATP-dependent regulation. | ||||||||||||||||||
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_11356.map.gz | 129.6 MB | EMDB map data format | |
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Header (meta data) | emd-11356-v30.xml emd-11356.xml | 20.1 KB 20.1 KB | Display Display | EMDB header |
Images | emd_11356.png | 90 KB | ||
Others | emd_11356_additional_1.map.gz emd_11356_additional_2.map.gz emd_11356_additional_3.map.gz | 248 MB 162.6 MB 241.8 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-11356 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-11356 | HTTPS FTP |
-Validation report
Summary document | emd_11356_validation.pdf.gz | 235.4 KB | Display | EMDB validaton report |
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Full document | emd_11356_full_validation.pdf.gz | 234.6 KB | Display | |
Data in XML | emd_11356_validation.xml.gz | 7.5 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-11356 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-11356 | HTTPS FTP |
-Related structure data
Related structure data | 6yarC 6yayC 6yb3C 6yb5C 6ybbC 6ybuC 6yg8C C: citing same article (ref.) |
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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_11356.map.gz / Format: CCP4 / Size: 262.9 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | Main map non-uniformly refined. It represents two BcsB octamers (6YG8) linked by a central miscellaneous structure. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.13 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Additional map: Main map non-uniformly refined and sharpened.
File | emd_11356_additional_1.map | ||||||||||||
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Annotation | Main map non-uniformly refined and sharpened. | ||||||||||||
Projections & Slices |
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Density Histograms |
-Additional map: The volumes corresponding to the octamers were extracted...
File | emd_11356_additional_2.map | ||||||||||||
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Annotation | The volumes corresponding to the octamers were extracted and locally refined to be finally combined resulting in a map without the central miscellaneous structure. | ||||||||||||
Projections & Slices |
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Density Histograms |
-Additional map: The volumes corresponding to the octamers were extracted...
File | emd_11356_additional_3.map | ||||||||||||
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Annotation | The volumes corresponding to the octamers were extracted and locally refined to be finally combined and sharpened resulting in a map without the central miscellaneous structure. | ||||||||||||
Projections & Slices |
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Density Histograms |
-Sample components
-Entire : Self-assembly of the BcsB protein
Entire | Name: Self-assembly of the BcsB protein |
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Components |
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-Supramolecule #1: Self-assembly of the BcsB protein
Supramolecule | Name: Self-assembly of the BcsB protein / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
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Source (natural) | Organism: Escherichia coli (E. coli) / Strain: 1094 / Location in cell: periplasm |
Recombinant expression | Organism: Escherichia coli BL21(DE3) (bacteria) / Recombinant plasmid: pET-21b |
Molecular weight | Theoretical: 1.378 MDa |
-Macromolecule #1: Bacterial cellulose synthase regulator protein BcsB
Macromolecule | Name: Bacterial cellulose synthase regulator protein BcsB / type: protein_or_peptide / ID: 1 Details: BcsB harbors a signal sequence to be addressed to the periplasm where according the ServerP4.1 Server prediction would result in mature form starting with the residue #26 TPATQ... Enantiomer: LEVO |
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Source (natural) | Organism: Escherichia coli (E. coli) |
Recombinant expression | Organism: Escherichia coli BL21(DE3) (bacteria) |
Sequence | String: TPATQPLINA EPAVAAQTEQ NPQVGQVMPG VQGADAPVVA QNGPSRDVKL TFAQIAPPPG SMVLRGINPN GSIEFGMRSD EVVTKAMLNL EYTPS PSLL PVQSQLKVYL NDELMGVLPV TKEQLGKKTL AQMPINPLFI TDFNRVRLEF VGHYQD VCE NPASTTLWLD ...String: TPATQPLINA EPAVAAQTEQ NPQVGQVMPG VQGADAPVVA QNGPSRDVKL TFAQIAPPPG SMVLRGINPN GSIEFGMRSD EVVTKAMLNL EYTPS PSLL PVQSQLKVYL NDELMGVLPV TKEQLGKKTL AQMPINPLFI TDFNRVRLEF VGHYQD VCE NPASTTLWLD VGRSSGLDLT YQTLNVKNDL SHFPVPFFDP RDNRTNTLPM VFAGAPD VG LQQASAIVAS WFGSRSGWRG QNFPVLYNQL PDRNAIVFAT NDKRPDFLRD HPAVKAPV I EMINHPQNPY VKLLVVFGRD DKDLLQAAKG IAQGNILFRG ESVVVNEVKP LLPRKPYDA PNWVRTDRPV TFGELKTYEE QLQSSGLEPA AINVSLNLPP DLYLMRSTGI DMDINYRYTM PPVKDSSRM DISLNNQFLQ SFNLSSKQEA NRLLLRIPVL QGLLDGKTDV SIPALKLGAT N QLRFDFEY MNPMPGGSVD NCITFQPVQN HVVIGDDSTI DFSKYYHFIP MPDLRAFANA GF PFSRMAD LSQTITVMPK APNEAQMETL LNTVGFIGAQ TGFPAINLTV TDDGSTIQGK DAD IMIIGG IPDKLKDDKQ IDLLVQATES WVKTPMRQTP FPGIVPDESD RAAETRSTLT SSGA MAAVI GFQSPYNDQR SVIALLADSP RGYEMLNDAV NDSGKRATMF GSVAVIRESG INSLR VGDV YYVGHLPWFE RLWYALANHP ILLAVLAAIS VILLAWVLWR LLRIISRRRL NPDNEAAALE HHHHHH |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 8 Component:
Details: BcsB was incubated with a mix of detergents: 0.4% DDM, 0.4 % digitonin, 0.4% DM-NPG, 0.2 % GDN-101, and 0.2% LM-NPG and then purified on IMAC using free-detergents buffers. | ||||||||
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Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK IV |
-Electron microscopy
Microscope | TFS TALOS |
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Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Average electron dose: 0.74 e/Å2 |
Electron beam | Acceleration voltage: 200 kV / Electron source: FIELD EMISSION GUN |
Electron optics | C2 aperture diameter: 100.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 2.75 µm / Nominal defocus min: 0.75 µm / Nominal magnification: 36000 |