+Open data
-Basic information
Entry | Database: PDB / ID: 7l2z | ||||||||||||
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Title | Bacterial cellulose synthase BcsB hexamer | ||||||||||||
Components | Cyclic di-GMP-binding protein | ||||||||||||
Keywords | BIOSYNTHETIC PROTEIN / bacterial cellulose / periplasmic / structural subunit | ||||||||||||
Function / homology | Cellulose synthase, subunit B / Cellulose synthase BcsB, bacterial / Bacterial cellulose synthase subunit / cellulose biosynthetic process / UDP-alpha-D-glucose metabolic process / plasma membrane / Cyclic di-GMP-binding protein Function and homology information | ||||||||||||
Biological species | Escherichia coli (E. coli) | ||||||||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.4 Å | ||||||||||||
Authors | Acheson, J.F. / Zimmer, J. | ||||||||||||
Funding support | United States, 3items
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Citation | Journal: Nat Struct Mol Biol / Year: 2021 Title: Molecular organization of the E. coli cellulose synthase macrocomplex. Authors: Justin F Acheson / Ruoya Ho / Nicolette F Goularte / Lynette Cegelski / Jochen Zimmer / Abstract: Cellulose is frequently found in communities of sessile bacteria called biofilms. Escherichia coli and other enterobacteriaceae modify cellulose with phosphoethanolamine (pEtN) to promote host tissue ...Cellulose is frequently found in communities of sessile bacteria called biofilms. Escherichia coli and other enterobacteriaceae modify cellulose with phosphoethanolamine (pEtN) to promote host tissue adhesion. The E. coli pEtN cellulose biosynthesis machinery contains the catalytic BcsA-B complex that synthesizes and secretes cellulose, in addition to five other subunits. The membrane-anchored periplasmic BcsG subunit catalyzes pEtN modification. Here we present the structure of the roughly 1 MDa E. coli Bcs complex, consisting of one BcsA enzyme associated with six copies of BcsB, determined by single-particle cryo-electron microscopy. BcsB homo-oligomerizes primarily through interactions between its carbohydrate-binding domains as well as intermolecular beta-sheet formation. The BcsB hexamer creates a half spiral whose open side accommodates two BcsG subunits, directly adjacent to BcsA's periplasmic channel exit. The cytosolic BcsE and BcsQ subunits associate with BcsA's regulatory PilZ domain. The macrocomplex is a fascinating example of cellulose synthase specification. | ||||||||||||
History |
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-Structure visualization
Movie |
Movie viewer |
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Structure viewer | Molecule: MolmilJmol/JSmol |
-Downloads & links
-Download
PDBx/mmCIF format | 7l2z.cif.gz | 1.2 MB | Display | PDBx/mmCIF format |
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PDB format | pdb7l2z.ent.gz | 1 MB | Display | PDB format |
PDBx/mmJSON format | 7l2z.json.gz | Tree view | PDBx/mmJSON format | |
Others | Other downloads |
-Validation report
Summary document | 7l2z_validation.pdf.gz | 902.1 KB | Display | wwPDB validaton report |
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Full document | 7l2z_full_validation.pdf.gz | 915.6 KB | Display | |
Data in XML | 7l2z_validation.xml.gz | 98.8 KB | Display | |
Data in CIF | 7l2z_validation.cif.gz | 153.3 KB | Display | |
Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/l2/7l2z ftp://data.pdbj.org/pub/pdb/validation_reports/l2/7l2z | HTTPS FTP |
-Related structure data
Related structure data | 23146MC 7lbyC M: map data used to model this data C: citing same article (ref.) |
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Similar structure data | |
EM raw data | EMPIAR-10626 (Title: Cryo EM Structure of the E. coli BcsB Hexamer / Data size: 837.2 Data #1: Unaligned multi-frame micrographs [micrographs - multiframe] Data #2: Unaligned movie frames [micrographs - multiframe] / Data #3: Unaligned movie frames [micrographs - multiframe] / Data #4: Unaligned movie frames [micrographs - multiframe]) EMPIAR-10627 (Title: Poly-alanine backbone model of E. coli BcsA bound to BcsB Data size: 3.2 TB Data #1: Unaligned multi-frame micrographs [micrographs - multiframe] Data #2: Unaligned movie frames [micrographs - multiframe] / Data #3: Unaligned movie frames [micrographs - multiframe] / Data #4: Unaligned movie frames [micrographs - multiframe] / Data #5: Unaligned movie frames [micrographs - multiframe]) |
-Links
-Assembly
Deposited unit |
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-Components
#1: Protein | Mass: 84304.906 Da / Num. of mol.: 6 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Escherichia coli (strain K12) (bacteria) Strain: K12 / Gene: bcsB, yhjN, b3532, JW3500 / Plasmid: petduet_BcsA_BcsB_AdrA / Production host: Escherichia coli BL21 (bacteria) / Strain (production host): BL21(DE3) / Variant (production host): C43 / References: UniProt: P37652 |
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-Experimental details
-Experiment
Experiment | Method: ELECTRON MICROSCOPY |
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EM experiment | Aggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction |
-Sample preparation
Component | Name: Bacterial cellulose synthase BcsB hexamer / Type: COMPLEX Details: Periplasmic domain of BcsB refined using masked local refinement and signal subtraction and from E coli BCS inner-membrane complex projections. Entity ID: all / Source: RECOMBINANT | ||||||||||||||||||||||||||||||||||||||||
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Molecular weight | Value: 0.481 MDa / Experimental value: NO | ||||||||||||||||||||||||||||||||||||||||
Source (natural) | Organism: Escherichia coli K-12 (bacteria) / Cellular location: periplasm innermembrane | ||||||||||||||||||||||||||||||||||||||||
Source (recombinant) | Organism: Escherichia coli BL21(DE3) (bacteria) / Strain: C43 / Plasmid: petduet_BcsA_BcsB_AdrA | ||||||||||||||||||||||||||||||||||||||||
Buffer solution | pH: 8 Details: All buffers made fresh from concentrated stock. A 10% LMNG 2% DMNG 2% CHS solution was made made in milliQ water without buffer. After rocking for one day at room temp the solution was ...Details: All buffers made fresh from concentrated stock. A 10% LMNG 2% DMNG 2% CHS solution was made made in milliQ water without buffer. After rocking for one day at room temp the solution was briefly sonicated to completely dissolve the CHS. | ||||||||||||||||||||||||||||||||||||||||
Buffer component |
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Specimen | Conc.: 0.4 mg/ml / Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES | ||||||||||||||||||||||||||||||||||||||||
Specimen support | Details: 2 drops of amylamine were added to the chamber / Grid material: COPPER / Grid mesh size: 300 divisions/in. / Grid type: C-flat-1.2/1.3 | ||||||||||||||||||||||||||||||||||||||||
Vitrification | Instrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE / Humidity: 100 % / Chamber temperature: 277 K Details: 3.5 uL applied to c-flat 1.2/1.3 300 mesh grids incubated for 30s then blotted using force 6 for 12 seconds |
-Electron microscopy imaging
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
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Microscopy | Model: FEI TITAN KRIOS |
Electron gun | Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM |
Electron lens | Mode: BRIGHT FIELD / Nominal magnification: 81000 X / Nominal defocus max: -2500 nm / Nominal defocus min: -1000 nm / Cs: 2.7 mm |
Specimen holder | Cryogen: NITROGEN / Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER |
Image recording | Electron dose: 51 e/Å2 / Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Num. of grids imaged: 1 / Num. of real images: 2964 / Details: movie mode 40 frames |
EM imaging optics | Energyfilter name: GIF Bioquantum / Energyfilter slit width: 10 eV |
-Processing
Software | Name: PHENIX / Version: 1.17.1_3660: / Classification: refinement | |||||||||||||||||||||||||||||||||||
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EM software |
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CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | |||||||||||||||||||||||||||||||||||
Particle selection | Num. of particles selected: 409611 | |||||||||||||||||||||||||||||||||||
3D reconstruction | Resolution: 3.4 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 115289 / Num. of class averages: 1 / Symmetry type: POINT | |||||||||||||||||||||||||||||||||||
Atomic model building | B value: 101.45 / Protocol: AB INITIO MODEL / Space: REAL / Target criteria: CC | |||||||||||||||||||||||||||||||||||
Refine LS restraints |
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