[English] 日本語
![](img/lk-miru.gif)
- PDB-7qgq: Extended H/L (SLPH/SLPL) complex from C. difficile (CD630 strain)... -
+
Open data
-
Basic information
Entry | Database: PDB / ID: 7qgq | ||||||
---|---|---|---|---|---|---|---|
Title | Extended H/L (SLPH/SLPL) complex from C. difficile (CD630 strain) fit into R20291 S-layer negative stain map | ||||||
![]() | (Precursor of the S-layer proteins) x 2 | ||||||
![]() | STRUCTURAL PROTEIN / Bacterial surface / S-layer | ||||||
Function / homology | : / Low molecular weight S layer protein, N-terminal / Low molecular weight S layer protein, N-terminal, subdomain / Low molecular weight S layer protein N terminal / Putative cell wall binding repeat 2 / Cell wall binding domain 2 (CWB2) / outer membrane-bounded periplasmic space / identical protein binding / Precursor of the S-layer proteins![]() | ||||||
Biological species | ![]() | ||||||
Method | ELECTRON CRYSTALLOGRAPHY / electron crystallography / negative staining | ||||||
![]() | Banerji, O. / Wilson, J.S. / Bullough, P.A. | ||||||
Funding support | ![]()
| ||||||
![]() | ![]() Title: Structure and assembly of the S-layer in C. difficile. Authors: Paola Lanzoni-Mangutchi / Oishik Banerji / Jason Wilson / Anna Barwinska-Sendra / Joseph A Kirk / Filipa Vaz / Shauna O'Beirne / Arnaud Baslé / Kamel El Omari / Armin Wagner / Neil F ...Authors: Paola Lanzoni-Mangutchi / Oishik Banerji / Jason Wilson / Anna Barwinska-Sendra / Joseph A Kirk / Filipa Vaz / Shauna O'Beirne / Arnaud Baslé / Kamel El Omari / Armin Wagner / Neil F Fairweather / Gillian R Douce / Per A Bullough / Robert P Fagan / Paula S Salgado / ![]() ![]() Abstract: Many bacteria and archaea possess a two-dimensional protein array, or S-layer, that covers the cell surface and plays crucial roles in cell physiology. Here, we report the crystal structure of SlpA, ...Many bacteria and archaea possess a two-dimensional protein array, or S-layer, that covers the cell surface and plays crucial roles in cell physiology. Here, we report the crystal structure of SlpA, the main S-layer protein of the bacterial pathogen Clostridioides difficile, and use electron microscopy to study S-layer organisation and assembly. The SlpA crystal lattice mimics S-layer assembly in the cell, through tiling of triangular prisms above the cell wall, interlocked by distinct ridges facing the environment. Strikingly, the array is very compact, with pores of only ~10 Å in diameter, compared to other S-layers (30-100 Å). The surface-exposed flexible ridges are partially dispensable for overall structure and assembly, although a mutant lacking this region becomes susceptible to lysozyme, an important molecule in host defence. Thus, our work gives insights into S-layer organisation and provides a basis for development of C. difficile-specific therapeutics. | ||||||
History |
|
-
Structure visualization
Structure viewer | Molecule: ![]() ![]() |
---|
-
Downloads & links
-
Download
PDBx/mmCIF format | ![]() | 2.6 MB | Display | ![]() |
---|---|---|---|---|
PDB format | ![]() | Display | ![]() | |
PDBx/mmJSON format | ![]() | Tree view | ![]() | |
Others | ![]() |
-Validation report
Arichive directory | ![]() ![]() | HTTPS FTP |
---|
-Related structure data
Related structure data | ![]() 13957MC ![]() 7acvC ![]() 7acwC ![]() 7acxC ![]() 7acyC ![]() 7aczC M: map data used to model this data C: citing same article ( |
---|---|
Similar structure data | Similarity search - Function & homology ![]() |
-
Links
-
Assembly
Deposited unit | ![]()
| ||||||||
---|---|---|---|---|---|---|---|---|---|
1 |
| ||||||||
Unit cell |
|
-
Components
#1: Protein | Mass: 39495.035 Da / Num. of mol.: 12 / Source method: isolated from a natural source / Source: (natural) ![]() #2: Protein | Mass: 33949.785 Da / Num. of mol.: 12 / Source method: isolated from a natural source / Source: (natural) ![]() |
---|
-Experimental details
-Experiment
Experiment | Method: ELECTRON CRYSTALLOGRAPHY |
---|---|
EM experiment | Aggregation state: 2D ARRAY / 3D reconstruction method: electron crystallography |
-
Sample preparation
Component | Name: Electron crystallographic reconstruction of R20291 S-layer, extended to cover 12 molecules of SlpA for flexible fitting of X-ray structure to map Type: COMPLEX / Entity ID: all / Source: NATURAL |
---|---|
Molecular weight | Experimental value: NO |
Source (natural) | Organism: ![]() |
Buffer solution | pH: 7.4 |
Specimen | Embedding applied: NO / Shadowing applied: NO / Staining applied: YES / Vitrification applied: NO |
EM staining | Type: NEGATIVE Details: Negatively stained EM samples were prepared by depositing sample on continuous carbon layer and staining with 2 % Uranyl Formate with blotting Material: Uranyl Formate |
Specimen support | Grid material: COPPER / Grid mesh size: 300 divisions/in. / Grid type: Homemade |
-Data collection
Microscopy | Model: FEI/PHILIPS CM200FEG |
---|---|
Electron gun | Electron source: ![]() |
Electron lens | Mode: BRIGHT FIELD / Nominal defocus max: 2200 nm / Nominal defocus min: 800 nm |
Image recording | Electron dose: 0.1 e/Å2 / Film or detector model: GATAN ULTRASCAN 4000 (4k x 4k) / Num. of real images: 36 / Details: Images binned by 2 before processing |
Image scans | Sampling size: 15 µm / Width: 2048 / Height: 2048 |
EM diffraction | Camera length: 0.1 mm |
EM diffraction shell | Resolution: 18.25→78.79 Å / Fourier space coverage: 0.1 % / Multiplicity: 0.1 / Num. of structure factors: 710 / Phase residual: 21.9 ° |
EM diffraction stats | Fourier space coverage: 0.1 % / High resolution: 18.25 Å / Num. of intensities measured: 1085 / Num. of structure factors: 710 / Phase error: 22.3 ° / Phase residual: 21.9 ° Phase error rejection criteria: Structure factors with phase errors higher than 45 degrees were omitted from refinement Rmerge: 0.193 |
-
Processing
EM software |
| ||||||||||||||||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
EM 3D crystal entity | ∠α: 90 ° / ∠β: 90 ° / ∠γ: 100 ° / A: 80 Å / B: 80 Å / C: 160 Å / Space group name: P112 / Space group num: 13 | ||||||||||||||||||||||||||||||
CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||||||||
3D reconstruction | Resolution method: DIFFRACTION PATTERN/LAYERLINES / Symmetry type: 3D CRYSTAL | ||||||||||||||||||||||||||||||
Atomic model building | Protocol: FLEXIBLE FIT / Space: REAL | ||||||||||||||||||||||||||||||
Refinement | Cross valid method: THROUGHOUT | ||||||||||||||||||||||||||||||
Displacement parameters | Biso max: 104.01 Å2 / Biso mean: 57.857 Å2 / Biso min: 34.65 Å2 |