+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-12651 | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Title | cryoEM structure of 2C9-sMAC | ||||||||||||
Map data | Locally sharpened 2C9-sMAC map, resolution 3.27 A | ||||||||||||
Sample |
| ||||||||||||
Keywords | Complement / MACPF / Membrane Attack Complex / CDC / pore forming / IMMUNE SYSTEM | ||||||||||||
Function / homology | Function and homology information cell killing / Terminal pathway of complement / membrane attack complex / complement binding / other organism cell membrane / Activation of C3 and C5 / negative regulation of macrophage chemotaxis / complement activation / complement activation, alternative pathway / chemokine activity ...cell killing / Terminal pathway of complement / membrane attack complex / complement binding / other organism cell membrane / Activation of C3 and C5 / negative regulation of macrophage chemotaxis / complement activation / complement activation, alternative pathway / chemokine activity / retinol binding / endopeptidase inhibitor activity / positive regulation of vascular endothelial growth factor production / complement activation, classical pathway / positive regulation of chemokine production / Peptide ligand-binding receptors / Regulation of Complement cascade / protein homooligomerization / chemotaxis / positive regulation of immune response / extracellular vesicle / G alpha (i) signalling events / in utero embryonic development / blood microparticle / killing of cells of another organism / cell surface receptor signaling pathway / inflammatory response / immune response / G protein-coupled receptor signaling pathway / innate immune response / signaling receptor binding / protein-containing complex binding / extracellular space / extracellular exosome / extracellular region / membrane / plasma membrane Similarity search - Function | ||||||||||||
Biological species | Homo sapiens (human) | ||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.27 Å | ||||||||||||
Authors | Menny A / Couves EC | ||||||||||||
Funding support | European Union, United Kingdom, 3 items
| ||||||||||||
Citation | Journal: Nat Commun / Year: 2021 Title: Structural basis of soluble membrane attack complex packaging for clearance. Authors: Anaïs Menny / Marie V Lukassen / Emma C Couves / Vojtech Franc / Albert J R Heck / Doryen Bubeck / Abstract: Unregulated complement activation causes inflammatory and immunological pathologies with consequences for human disease. To prevent bystander damage during an immune response, extracellular ...Unregulated complement activation causes inflammatory and immunological pathologies with consequences for human disease. To prevent bystander damage during an immune response, extracellular chaperones (clusterin and vitronectin) capture and clear soluble precursors to the membrane attack complex (sMAC). However, how these chaperones block further polymerization of MAC and prevent the complex from binding target membranes remains unclear. Here, we address that question by combining cryo electron microscopy (cryoEM) and cross-linking mass spectrometry (XL-MS) to solve the structure of sMAC. Together our data reveal how clusterin recognizes and inhibits polymerizing complement proteins by binding a negatively charged surface of sMAC. Furthermore, we show that the pore-forming C9 protein is trapped in an intermediate conformation whereby only one of its two transmembrane β-hairpins has unfurled. This structure provides molecular details for immune pore formation and helps explain a complement control mechanism that has potential implications for how cell clearance pathways mediate immune homeostasis. | ||||||||||||
History |
|
-Structure visualization
Movie |
Movie viewer |
---|---|
Structure viewer | EM map: SurfViewMolmilJmol/JSmol |
Supplemental images |
-Downloads & links
-EMDB archive
Map data | emd_12651.map.gz | 216.9 MB | EMDB map data format | |
---|---|---|---|---|
Header (meta data) | emd-12651-v30.xml emd-12651.xml | 29.3 KB 29.3 KB | Display Display | EMDB header |
Images | emd_12651.png | 54 KB | ||
Filedesc metadata | emd-12651.cif.gz | 9.7 KB | ||
Others | emd_12651_half_map_1.map.gz emd_12651_half_map_2.map.gz | 193.7 MB 193.7 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-12651 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-12651 | HTTPS FTP |
-Validation report
Summary document | emd_12651_validation.pdf.gz | 733.8 KB | Display | EMDB validaton report |
---|---|---|---|---|
Full document | emd_12651_full_validation.pdf.gz | 733.4 KB | Display | |
Data in XML | emd_12651_validation.xml.gz | 15.9 KB | Display | |
Data in CIF | emd_12651_validation.cif.gz | 19 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-12651 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-12651 | HTTPS FTP |
-Related structure data
Related structure data | 7nydMC 7nycC C: citing same article (ref.) M: atomic model generated by this map |
---|---|
Similar structure data |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
---|---|
Related items in Molecule of the Month |
-Map
File | Download / File: emd_12651.map.gz / Format: CCP4 / Size: 244.1 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Annotation | Locally sharpened 2C9-sMAC map, resolution 3.27 A | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.047 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
|
-Supplemental data
-Half map: Half map 2 from the 3D auto-refinement of 2C9-sMAC
File | emd_12651_half_map_1.map | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Annotation | Half map 2 from the 3D auto-refinement of 2C9-sMAC | ||||||||||||
Projections & Slices |
| ||||||||||||
Density Histograms |
-Half map: Half map 1 from the 3D auto-refinement of 2C9-sMAC
File | emd_12651_half_map_2.map | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Annotation | Half map 1 from the 3D auto-refinement of 2C9-sMAC | ||||||||||||
Projections & Slices |
| ||||||||||||
Density Histograms |
-Sample components
+Entire : 2C9-sMAC
+Supramolecule #1: 2C9-sMAC
+Macromolecule #1: Complement component C8 beta chain
+Macromolecule #2: Complement component C8 alpha chain
+Macromolecule #3: Complement component C9
+Macromolecule #4: Complement component C7
+Macromolecule #5: Complement component C6
+Macromolecule #6: Complement component C8 gamma chain
+Macromolecule #7: Complement C5
+Macromolecule #11: alpha-D-mannopyranose
+Macromolecule #12: CALCIUM ION
+Macromolecule #13: 2-acetamido-2-deoxy-beta-D-glucopyranose
+Macromolecule #14: alpha-L-fucopyranose
-Experimental details
-Structure determination
Method | cryo EM |
---|---|
Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Concentration | 0.065 mg/mL |
---|---|
Buffer | pH: 7.4 |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 95 % / Chamber temperature: 295 K / Instrument: FEI VITROBOT MARK III |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
---|---|
Image recording | Film or detector model: GATAN K2 QUANTUM (4k x 4k) / Detector mode: COUNTING / Average electron dose: 40.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | C2 aperture diameter: 70.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
-Image processing
Startup model | Type of model: INSILICO MODEL In silico model: The initial model was generated in Relion 3.1 |
---|---|
Final reconstruction | Resolution.type: BY AUTHOR / Resolution: 3.27 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: RELION (ver. 3.1) / Number images used: 142499 |
Initial angle assignment | Type: MAXIMUM LIKELIHOOD / Software - Name: RELION (ver. 3.1) |
Final angle assignment | Type: MAXIMUM LIKELIHOOD / Software - Name: RELION (ver. 3.1) |
-Atomic model buiding 1
Initial model |
| ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Refinement | Protocol: RIGID BODY FIT | ||||||||||||
Output model | PDB-7nyd: |