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- EMDB-2436: Cryo-EM reconstruction of empty enterovirus 71 in complex with a ... -
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Basic information
Entry | Database: EMDB / ID: EMD-2436 | |||||||||
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Title | Cryo-EM reconstruction of empty enterovirus 71 in complex with a neutralizing antibody E19 | |||||||||
![]() | Reconstruction of human enterovirus 71 with bound Fab fragments of antibody E19 | |||||||||
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![]() | EV71 / human / enterovirus 71 / virus / antibody / Fab / E19 / complex / neutralizing / pathogen / genome / release | |||||||||
Function / homology | ![]() symbiont-mediated suppression of host cytoplasmic pattern recognition receptor signaling pathway via inhibition of MDA-5 activity / picornain 2A / symbiont-mediated suppression of host mRNA export from nucleus / symbiont genome entry into host cell via pore formation in plasma membrane / picornain 3C / T=pseudo3 icosahedral viral capsid / host cell cytoplasmic vesicle membrane / nucleoside-triphosphate phosphatase / channel activity / monoatomic ion transmembrane transport ...symbiont-mediated suppression of host cytoplasmic pattern recognition receptor signaling pathway via inhibition of MDA-5 activity / picornain 2A / symbiont-mediated suppression of host mRNA export from nucleus / symbiont genome entry into host cell via pore formation in plasma membrane / picornain 3C / T=pseudo3 icosahedral viral capsid / host cell cytoplasmic vesicle membrane / nucleoside-triphosphate phosphatase / channel activity / monoatomic ion transmembrane transport / DNA replication / RNA helicase activity / endocytosis involved in viral entry into host cell / symbiont-mediated activation of host autophagy / RNA-directed RNA polymerase / cysteine-type endopeptidase activity / viral RNA genome replication / RNA-directed RNA polymerase activity / DNA-templated transcription / virion attachment to host cell / host cell nucleus / structural molecule activity / ATP hydrolysis activity / proteolysis / RNA binding / zinc ion binding / ATP binding / membrane Similarity search - Function | |||||||||
Biological species | ![]() ![]() ![]() ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 13.0 Å | |||||||||
![]() | Plevka P / Perera R / Cardosa J / Suksatu A / Kuhn RJ / Rossmann MG | |||||||||
![]() | ![]() Title: Neutralizing antibodies can initiate genome release from human enterovirus 71. Authors: Pavel Plevka / Pei-Yin Lim / Rushika Perera / Jane Cardosa / Ampa Suksatu / Richard J Kuhn / Michael G Rossmann / ![]() Abstract: Antibodies were prepared by immunizing mice with empty, immature particles of human enterovirus 71 (EV71), a picornavirus that causes severe neurological disease in young children. The capsid ...Antibodies were prepared by immunizing mice with empty, immature particles of human enterovirus 71 (EV71), a picornavirus that causes severe neurological disease in young children. The capsid structure of these empty particles is different from that of the mature virus and is similar to "A" particles encountered when picornaviruses recognize a potential host cell before genome release. The monoclonal antibody E18, generated by this immunization, induced a conformational change when incubated at temperatures between 4 °C and 37 °C with mature virus, transforming infectious virions into A particles. The resultant loss of genome that was observed by cryo-EM and a fluorescent SYBR Green dye assay inactivated the virus, establishing the mechanism by which the virus is inactivated and demonstrating that the E18 antibody has potential as an anti-EV71 therapy. The antibody-mediated virus neutralization by the induction of genome release has not been previously demonstrated. Furthermore, the present results indicate that antibodies with genome-release activity could also be produced for other picornaviruses by immunization with immature particles. | |||||||||
History |
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Structure visualization
Movie |
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Structure viewer | EM map: ![]() ![]() ![]() |
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 23.9 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 12.3 KB 12.3 KB | Display Display | ![]() |
Images | ![]() | 228.4 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 4c10MC ![]() 2397C ![]() 2434C ![]() 4c0uC ![]() 4c0yC M: atomic model generated by this map C: citing same article ( |
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Similar structure data |
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Links
EMDB pages | ![]() ![]() |
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Related items in Molecule of the Month |
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Map
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Annotation | Reconstruction of human enterovirus 71 with bound Fab fragments of antibody E19 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 2.48 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
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Sample components
-Entire : Reconstruction of human enterovirus 71 with bound Fab fragments o...
Entire | Name: Reconstruction of human enterovirus 71 with bound Fab fragments of antibody E19 |
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Components |
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-Supramolecule #1000: Reconstruction of human enterovirus 71 with bound Fab fragments o...
Supramolecule | Name: Reconstruction of human enterovirus 71 with bound Fab fragments of antibody E19 type: sample / ID: 1000 Oligomeric state: Icosahedral virus and 60 copies of bound Fab Number unique components: 2 |
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Molecular weight | Theoretical: 6.8 MDa |
-Supramolecule #1: Human enterovirus 71
Supramolecule | Name: Human enterovirus 71 / type: virus / ID: 1 / Name.synonym: EV71 / NCBI-ID: 39054 / Sci species name: Human enterovirus 71 / Virus type: VIRION / Virus isolate: SEROTYPE / Virus enveloped: No / Virus empty: Yes / Syn species name: EV71 / Sci species serotype: EV-174 |
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Host (natural) | Organism: ![]() |
Molecular weight | Theoretical: 3.2 MDa |
Virus shell | Shell ID: 1 / Name: capsid / Diameter: 280 Å / T number (triangulation number): 1 |
-Macromolecule #1: EV19/5/C1-6/F1/C11
Macromolecule | Name: EV19/5/C1-6/F1/C11 / type: protein_or_peptide / ID: 1 / Name.synonym: E19 / Number of copies: 60 / Oligomeric state: monomer / Recombinant expression: No |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 4.6 KDa |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Concentration | 2 mg/mL |
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Buffer | pH: 8 / Details: 10 mM Tris-HCl, pH 8.0, 0.5 M NaCl, and 1 mM EDTA |
Grid | Details: Quantifoin R1.2/1.3 Holey carbon 200Mesh |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 70 % / Chamber temperature: 100 K / Instrument: HOMEMADE PLUNGER / Method: Blot for 2 second before plunging |
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Electron microscopy
Microscope | FEI/PHILIPS CM200FEG |
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Temperature | Min: 95 K / Max: 120 K / Average: 119 K |
Alignment procedure | Legacy - Astigmatism: Objective lens astigmatism was corrected at 100,000 times magnification |
Date | Sep 16, 2013 |
Image recording | Category: FILM / Film or detector model: KODAK SO-163 FILM / Digitization - Scanner: NIKON SUPER COOLSCAN 9000 / Number real images: 142 / Average electron dose: 18 e/Å2 / Bits/pixel: 16 |
Electron beam | Acceleration voltage: 200 kV / Electron source: ![]() |
Electron optics | Calibrated magnification: 49500 / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.0 mm / Nominal defocus max: 4.5 µm / Nominal defocus min: 0.75 µm / Nominal magnification: 50000 |
Sample stage | Specimen holder model: GATAN LIQUID NITROGEN |
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Image processing
Details | Particles were boxed with e2boxer.py |
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CTF correction | Details: CTF determined for all particles from one film, phase flip |
Final reconstruction | Applied symmetry - Point group: I (icosahedral) / Algorithm: OTHER / Resolution.type: BY AUTHOR / Resolution: 13.0 Å / Resolution method: FSC 0.5 CUT-OFF / Software - Name: eman, eman2 / Number images used: 10833 |
Final two d classification | Number classes: 122 |
-Atomic model buiding 1
Initial model | PDB ID: Chain - #0 - Chain ID: A / Chain - #1 - Chain ID: B / Chain - #2 - Chain ID: C / Chain - #3 - Chain ID: D |
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Software | Name: EMFit |
Refinement | Space: REAL / Protocol: RIGID BODY FIT / Target criteria: Rcrit |
Output model | ![]() PDB-4c10: |