+Open data
-Basic information
Entry | Database: PDB / ID: 8tsh | ||||||
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Title | S. thermodepolymerans KpsMT(E151Q)-KpsE in complex with ATP | ||||||
Components |
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Keywords | MEMBRANE PROTEIN / ABC transporter / Capsular polysaccharide | ||||||
Function / homology | Function and homology information lipopolysaccharide transport / ABC-type transporter activity / ATP-binding cassette (ABC) transporter complex / protein tyrosine kinase activity / ATP hydrolysis activity / ATP binding / plasma membrane Similarity search - Function | ||||||
Biological species | Caldimonas thermodepolymerans (bacteria) | ||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.1 Å | ||||||
Authors | Kuklewicz, J. / Zimmer, J. | ||||||
Funding support | United States, 1items
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Citation | Journal: Nature / Year: 2024 Title: Molecular insights into capsular polysaccharide secretion. Authors: Jeremi Kuklewicz / Jochen Zimmer / Abstract: Capsular polysaccharides (CPSs) fortify the cell boundaries of many commensal and pathogenic bacteria. Through the ABC-transporter-dependent biosynthesis pathway, CPSs are synthesized intracellularly ...Capsular polysaccharides (CPSs) fortify the cell boundaries of many commensal and pathogenic bacteria. Through the ABC-transporter-dependent biosynthesis pathway, CPSs are synthesized intracellularly on a lipid anchor and secreted across the cell envelope by the KpsMT ABC transporter associated with the KpsE and KpsD subunits. Here we use structural and functional studies to uncover crucial steps of CPS secretion in Gram-negative bacteria. We show that KpsMT has broad substrate specificity and is sufficient for the translocation of CPSs across the inner bacterial membrane, and we determine the cell surface organization and localization of CPSs using super-resolution fluorescence microscopy. Cryo-electron microscopy analyses of the KpsMT-KpsE complex in six different states reveal a KpsE-encaged ABC transporter, rigid-body conformational rearrangements of KpsMT during ATP hydrolysis and recognition of a glycolipid inside a membrane-exposed electropositive canyon. In vivo CPS secretion assays underscore the functional importance of canyon-lining basic residues. Combined, our analyses suggest a molecular model of CPS secretion by ABC transporters. | ||||||
History |
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-Structure visualization
Structure viewer | Molecule: MolmilJmol/JSmol |
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-Downloads & links
-Download
PDBx/mmCIF format | 8tsh.cif.gz | 603.8 KB | Display | PDBx/mmCIF format |
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PDB format | pdb8tsh.ent.gz | 491.7 KB | Display | PDB format |
PDBx/mmJSON format | 8tsh.json.gz | Tree view | PDBx/mmJSON format | |
Others | Other downloads |
-Validation report
Summary document | 8tsh_validation.pdf.gz | 1.1 MB | Display | wwPDB validaton report |
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Full document | 8tsh_full_validation.pdf.gz | 1.1 MB | Display | |
Data in XML | 8tsh_validation.xml.gz | 76.3 KB | Display | |
Data in CIF | 8tsh_validation.cif.gz | 117 KB | Display | |
Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/ts/8tsh ftp://data.pdbj.org/pub/pdb/validation_reports/ts/8tsh | HTTPS FTP |
-Related structure data
Related structure data | 41592MC 8tsiC 8tslC 8tswC 8tt3C 8tunC 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: 26223.744 Da / Num. of mol.: 2 / Mutation: E151Q Source method: isolated from a genetically manipulated source Source: (gene. exp.) Caldimonas thermodepolymerans (bacteria) Gene: C1702_11080 / Production host: Escherichia coli (E. coli) / References: UniProt: A0A2S5T4B3 #2: Protein | Mass: 30800.959 Da / Num. of mol.: 2 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Caldimonas thermodepolymerans (bacteria) Gene: C1702_11075 / Production host: Escherichia coli (E. coli) / References: UniProt: A0A2S5T447 #3: Protein | Mass: 44057.266 Da / Num. of mol.: 8 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Caldimonas thermodepolymerans (bacteria) Gene: C1702_11085 / Production host: Escherichia coli (E. coli) / References: UniProt: A0A2S5T4A0 #4: Chemical | #5: Chemical | Has ligand of interest | Y | |
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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: ABC transporter KpsMT(E151Q) in complex with polysaccharide co-polymerase KpsE in ATP-bound state Type: COMPLEX / Entity ID: #1-#3 / Source: RECOMBINANT |
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Molecular weight | Experimental value: NO |
Source (natural) | Organism: Caldimonas thermodepolymerans (bacteria) |
Source (recombinant) | Organism: Escherichia coli (E. coli) |
Buffer solution | pH: 7.5 |
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: BRIGHT FIELD / Nominal defocus max: 1800 nm / Nominal defocus min: 1000 nm |
Image recording | Electron dose: 50 e/Å2 / Film or detector model: FEI FALCON III (4k x 4k) |
-Processing
CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||
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3D reconstruction | Resolution: 3.1 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 36731 / Symmetry type: POINT | ||||||||||||||||||||||||
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