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- EMDB-6702: Cryo-EM Study of an Engineered Enterovirus 71-Like Particle Mimic... -

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Entry
Database: EMDB / ID: 6702
TitleCryo-EM Study of an Engineered Enterovirus 71-Like Particle Mimicking 80S Empty Capsid
Map data
SampleEnterovirus A71:
virus
SourceEnterovirus A71
Methodsingle particle reconstruction / cryo EM / 3.71 Å resolution
AuthorsWang X / Ku Z / Zhang X / Cong Y / Huang Z
CitationJournal: J. Virol. / Year: 2018
Title: Structure, Immunogenicity, and Protective Mechanism of an Engineered Enterovirus 71-Like Particle Vaccine Mimicking 80S Empty Capsid.
Authors: Xiaoli Wang / Zhiqiang Ku / Xiang Zhang / Xiaohua Ye / Jinhuan Chen / Qingwei Liu / Wei Zhang / Chao Zhang / Zhenglin Fu / Xia Jin / Yao Cong / Zhong Huang
Abstract: Enterovirus 71 (EV71) is the major causative agent of severe hand, foot, and mouth disease, which affects millions of young children in the Asia-Pacific region annually. In this study, we engineered ...Enterovirus 71 (EV71) is the major causative agent of severe hand, foot, and mouth disease, which affects millions of young children in the Asia-Pacific region annually. In this study, we engineered a novel EV71 virus-like particle (VLP) that lacks VP4 (therefore designated VLP) and investigated its structure, antigenicity, and vaccine potential. The cryo-electron microscopy (cryo-EM) structure of VLP was reconstructed to 3.71-Å resolution. Results from structural and biochemical analyses revealed that VLP resembles the end product of the viral uncoating process, the 80S empty capsid. VLP is able to elicit high-titer neutralizing antibodies and to fully protect mice against lethal viral challenge. Mechanistic studies showed that, at the cellular level, the anti-VLP sera exert neutralization effects at both pre- and postattachment stages by inhibiting both virus attachment and internalization, and at the molecular level, the antisera can block multiple interactions between EV71 and its key receptors. Our study gives a better understanding of EV71 capsid assembly and provides important information for the design and development of new-generation vaccines for EV71, and perhaps for other enteroviruses, as well. Enterovirus 71 (EV71) infection may lead to severe hand, foot, and mouth disease, with significant morbidity and mortality. Knowledge regarding EV71 particle assembly remains limited. Here, we report the generation and characterization of a novel EV71 virus-like particle that lacks the VP4 capsid subunit protein. This particle, termed VLP, structurally mimics the 80S empty capsid, which is the end stage of EV71 uncoating. We further show that VLP exhibits desirable immunogenicity and protective efficacy in proof-of-concept studies. In addition, the inhibitory mechanisms of the VLP-induced antibodies are unraveled at both the cellular and molecular levels. Our work provides the first evidence of picornaviral particle assembly in the complete absence of VP4 and identifies VLP as an improved EV71 vaccine candidate with desirable traits. These findings not only enhance our understanding of particle assembly and uncoating of picornaviruses, but also provide important information for structure-guided vaccine design for EV71 and other enteroviruses.
DateDeposition: Feb 8, 2017 / Header (metadata) release: Oct 18, 2017 / Map release: Oct 25, 2017 / Last update: Nov 1, 2017

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Structure visualization

Movie
  • Surface view with section colored by density value
  • Surface level: 2.5
  • Imaged by UCSF Chimera
  • Download
  • Surface view colored by radius
  • Surface level: 2.5
  • Imaged by UCSF Chimera
  • Download
Movie viewer
Structure viewerEM map:
SurfViewMolmilJmol/JSmol
Supplemental images

Downloads & links

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Map

Fileemd_6702.map.gz (map file in CCP4 format, 322487 KB)
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
432 pix
1 Å/pix.
= 430.92 Å
432 pix
1 Å/pix.
= 430.92 Å
432 pix
1 Å/pix.
= 430.92 Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 0.9975 Å
Density
Contour Level:2.5 (by author), 2.5 (movie #1):
Minimum - Maximum-7.170642 - 13.352638
Average (Standard dev.)2.773026E-9 (1)
Details

EMDB XML:

Space Group Number1
Map Geometry
Axis orderXYZ
Dimensions432432432
Origin-216-216-216
Limit215215215
Spacing432432432
CellA=B=C: 430.92 Å
α=β=γ: 90 deg.

CCP4 map header:

modeImage stored as Reals
Å/pix. X/Y/Z0.99750.99750.9975
M x/y/z432432432
origin x/y/z0.0000.0000.000
length x/y/z430.920430.920430.920
α/β/γ90.00090.00090.000
start NX/NY/NZ
NX/NY/NZ
MAP C/R/S123
start NC/NR/NS-216-216-216
NC/NR/NS432432432
D min/max/mean-7.17113.3530.000

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Supplemental data

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Sample components

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Entire Enterovirus A71

EntireName: Enterovirus A71 / Number of components: 1

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Component #1: virus, Enterovirus A71

VirusName: Enterovirus A71 / Class: VIRUS-LIKE PARTICLE / Empty: Yes / Enveloped: No / Isolate: OTHER
SpeciesSpecies: Enterovirus A71
Source (engineered)Expression System: Spodoptera frugiperda (fall armyworm) / Vector: pIEX/Bac-1

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Experimental details

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Sample preparation

SpecimenSpecimen state: particle / Method: cryo EM
Sample solutionpH: 7
VitrificationInstrument: FEI VITROBOT MARK III / Cryogen name: ETHANE / Humidity: 100 %

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Electron microscopy imaging

Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company
ImagingMicroscope: FEI TITAN KRIOS
Electron gunElectron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Electron dose: 38 e/Å2 / Illumination mode: FLOOD BEAM
LensImaging mode: BRIGHT FIELD
Specimen HolderModel: OTHER
CameraDetector: GATAN K2 (4k x 4k)

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Image processing

ProcessingMethod: single particle reconstruction / Applied symmetry: I (icosahedral) / Number of projections: 7887
3D reconstructionResolution: 3.71 Å / Resolution method: FSC 0.143 CUT-OFF

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