+Open data
-Basic information
Entry | Database: PDB / ID: 8rfe | |||||||||||||||
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Title | CgsiGP2 sample in nanodisc | |||||||||||||||
Components | Cyclic beta 1-2 glucan synthetase | |||||||||||||||
Keywords | ANTIBIOTIC / cgs / cyclic glucan | |||||||||||||||
Function / homology | Function and homology information cellobiose phosphorylase / cellobiose phosphorylase activity / carbohydrate binding / carbohydrate metabolic process / membrane Similarity search - Function | |||||||||||||||
Biological species | Agrobacterium tumefaciens (bacteria) | |||||||||||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.8 Å | |||||||||||||||
Authors | Sedzicki, J. / Ni, D. / Lehmann, F. / Stahlberg, H. / Dehio, C. | |||||||||||||||
Funding support | Switzerland, 4items
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Citation | Journal: Nat Commun / Year: 2024 Title: Structure-function analysis of the cyclic β-1,2-glucan synthase from Agrobacterium tumefaciens. Authors: Jaroslaw Sedzicki / Dongchun Ni / Frank Lehmann / Henning Stahlberg / Christoph Dehio / Abstract: The synthesis of complex sugars is a key aspect of microbial biology. Cyclic β-1,2-glucan (CβG) is a circular polysaccharide critical for host interactions of many bacteria, including major ...The synthesis of complex sugars is a key aspect of microbial biology. Cyclic β-1,2-glucan (CβG) is a circular polysaccharide critical for host interactions of many bacteria, including major pathogens of humans (Brucella) and plants (Agrobacterium). CβG is produced by the cyclic glucan synthase (Cgs), a multi-domain membrane protein. So far, its structure as well as the mechanism underlining the synthesis have not been clarified. Here we use cryo-electron microscopy (cryo-EM) and functional approaches to study Cgs from A. tumefaciens. We determine the structure of this complex protein machinery and clarify key aspects of CβG synthesis, revealing a distinct mechanism that uses a tyrosine-linked oligosaccharide intermediate in cycles of polymerization and processing of the glucan chain. Our research opens possibilities for combating pathogens that rely on polysaccharide virulence factors and may lead to synthetic biology approaches for producing complex cyclic sugars. | |||||||||||||||
History |
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-Structure visualization
Structure viewer | Molecule: MolmilJmol/JSmol |
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-Downloads & links
-Download
PDBx/mmCIF format | 8rfe.cif.gz | 498.8 KB | Display | PDBx/mmCIF format |
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PDB format | pdb8rfe.ent.gz | 396.5 KB | Display | PDB format |
PDBx/mmJSON format | 8rfe.json.gz | Tree view | PDBx/mmJSON format | |
Others | Other downloads |
-Validation report
Summary document | 8rfe_validation.pdf.gz | 1.4 MB | Display | wwPDB validaton report |
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Full document | 8rfe_full_validation.pdf.gz | 1.5 MB | Display | |
Data in XML | 8rfe_validation.xml.gz | 85.5 KB | Display | |
Data in CIF | 8rfe_validation.cif.gz | 129.2 KB | Display | |
Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/rf/8rfe ftp://data.pdbj.org/pub/pdb/validation_reports/rf/8rfe | HTTPS FTP |
-Related structure data
Related structure data | 19118MC 8rf0C 8rf9C 8rfgC M: map data used to model this data C: citing same article (ref.) |
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Similar structure data | Similarity search - Function & homologyF&H Search |
-Links
-Assembly
Deposited unit |
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1 |
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-Components
#1: Protein | Mass: 313358.656 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Agrobacterium tumefaciens (bacteria) / References: UniProt: A0A0F4FQW4, cellobiose phosphorylase | ||
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#2: Polysaccharide | beta-D-glucopyranose-(1-2)-beta-D-glucopyranose-(1-2)-beta-D-glucopyranose-(1-2)-beta-D- ...beta-D-glucopyranose-(1-2)-beta-D-glucopyranose-(1-2)-beta-D-glucopyranose-(1-2)-beta-D-glucopyranose-(1-2)-beta-D-glucopyranose-(1-2)-beta-D-glucopyranose-(1-2)-beta-D-glucopyranose-(1-2)-beta-D-glucopyranose-(1-2)-beta-D-glucopyranose Type: oligosaccharide / Mass: 1477.282 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source | ||
#3: Chemical | Has ligand of interest | N | |
-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: cyclic b-1,2-glucan synthase from Agrobacterium tumefaciens Type: COMPLEX / Entity ID: #1 / Source: NATURAL |
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Source (natural) | Organism: Agrobacterium tumefaciens (bacteria) |
Buffer solution | pH: 8 |
Specimen | Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES |
Vitrification | Cryogen name: ETHANE |
-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: OTHER / Nominal defocus max: 2500 nm / Nominal defocus min: 1200 nm |
Image recording | Electron dose: 60 e/Å2 / Film or detector model: FEI FALCON IV (4k x 4k) |
-Processing
CTF correction | Type: NONE | ||||||||||||||||||||||||
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3D reconstruction | Resolution: 3.8 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 171473 / Algorithm: BACK PROJECTION / Symmetry type: POINT | ||||||||||||||||||||||||
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