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Open data
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Basic information
Entry | ![]() | |||||||||
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Title | 3C9, C5b9-CD59 cryoEM structure; focus refinement map | |||||||||
![]() | 3C9, C5b9-CD59 focus refine map. RELION locally filtered map | |||||||||
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Biological species | ![]() ![]() | |||||||||
Method | ![]() ![]() | |||||||||
![]() | Couves EC / Gardner S / Bubeck D | |||||||||
Funding support | European Union, ![]()
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![]() | ![]() Title: Structural basis for membrane attack complex inhibition by CD59. Authors: Emma C Couves / Scott Gardner / Tomas B Voisin / Jasmine K Bickel / Phillip J Stansfeld / Edward W Tate / Doryen Bubeck / ![]() Abstract: CD59 is an abundant immuno-regulatory receptor that protects human cells from damage during complement activation. Here we show how the receptor binds complement proteins C8 and C9 at the membrane to ...CD59 is an abundant immuno-regulatory receptor that protects human cells from damage during complement activation. Here we show how the receptor binds complement proteins C8 and C9 at the membrane to prevent insertion and polymerization of membrane attack complex (MAC) pores. We present cryo-electron microscopy structures of two inhibited MAC precursors known as C5b8 and C5b9. We discover that in both complexes, CD59 binds the pore-forming β-hairpins of C8 to form an intermolecular β-sheet that prevents membrane perforation. While bound to C8, CD59 deflects the cascading C9 β-hairpins, rerouting their trajectory into the membrane. Preventing insertion of C9 restricts structural transitions of subsequent monomers and indirectly halts MAC polymerization. We combine our structural data with cellular assays and molecular dynamics simulations to explain how the membrane environment impacts the dual roles of CD59 in controlling pore formation of MAC, and as a target of bacterial virulence factors which hijack CD59 to lyse human cells. | |||||||||
History |
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Structure visualization
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 334 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 16.8 KB 16.8 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 19.1 KB | Display | ![]() |
Images | ![]() | 54.6 KB | ||
Others | ![]() ![]() | 482.7 MB 482.8 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
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Links
EMDB pages | ![]() ![]() |
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Map
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Annotation | 3C9, C5b9-CD59 focus refine map. RELION locally filtered map | ||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 0.831 Å | ||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Half map: 3C9, C5b9-CD59 focus refine map. RELION unfiltered half map
File | emd_15783_half_map_1.map | ||||||||||||
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Annotation | 3C9, C5b9-CD59 focus refine map. RELION unfiltered half map | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: 3C9, C5b9-CD59 focus refine map. RELION unfiltered half map
File | emd_15783_half_map_2.map | ||||||||||||
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Annotation | 3C9, C5b9-CD59 focus refine map. RELION unfiltered half map | ||||||||||||
Projections & Slices |
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Density Histograms |
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Sample components
-Entire : 2C9, CD59 inhibited MAC Complex
Entire | Name: 2C9, CD59 inhibited MAC Complex |
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Components |
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-Supramolecule #1: 2C9, CD59 inhibited MAC Complex
Supramolecule | Name: 2C9, CD59 inhibited MAC Complex / type: complex / ID: 1 / Chimera: Yes / Parent: 0 / Macromolecule list: #1-#9 Details: Solved in a DOPC, MSP2N2 nanodisc with a myristolated cytotopic CD59. |
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Source (natural) | Organism: ![]() ![]() |
-Experimental details
-Structure determination
Method | ![]() |
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Aggregation state | particle |
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Sample preparation
Buffer | pH: 7.4 Component:
Details: 20 mM HEPES pH 7.4, 120 mM NaCl | |||||||||
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Grid | Model: Quantifoil R1.2/1.3 / Support film - Material: GRAPHENE OXIDE / Support film - topology: CONTINUOUS / Pretreatment - Type: PLASMA CLEANING / Pretreatment - Time: 60 sec. / Pretreatment - Atmosphere: AIR | |||||||||
Vitrification | Cryogen name: ETHANE / Chamber humidity: 95 % / Instrument: FEI VITROBOT MARK II |
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Electron microscopy
Microscope | FEI TITAN KRIOS |
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Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | C2 aperture diameter: 100.0 µm / Calibrated defocus max: 4.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD![]() |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER |
Image recording | Film or detector model: GATAN K3 (6k x 4k) / Number grids imaged: 4 / Number real images: 52838 / Average exposure time: 2.0 sec. / Average electron dose: 50.0 e/Å2 / Details: Collected over 4 separate data collections |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |