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- EMDB-6796: CryoEM structure of HPV18 L1-only VLP -

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Basic information

Entry
Database: EMDB / ID: 6796
TitleCryoEM structure of HPV18 L1-only VLP
Map data
SampleAlphapapillomavirus 7:
virus
SourceAlphapapillomavirus 7
Methodsingle particle reconstruction / cryo EM / 20 Å resolution
AuthorsLi ZH / Yan XD / Zheng QB / Gu Y / Li SW
CitationJournal: Vaccine / Year: 2017
Title: Characterization of an Escherichia coli-derived human papillomavirus type 16 and 18 bivalent vaccine.
Authors: Ying Gu / Minxi Wei / Daning Wang / Zhihai Li / Minghui Xie / Huirong Pan / Ting Wu / Jun Zhang / Shaowei Li / Ningshao Xia
Abstract: Human papillomavirus (HPV) types 16 and 18 account for approximately 70% of cervical cancer worldwide. Neutralizing HPV prophylactic vaccines offer significant benefit, as they block HPV infection ...Human papillomavirus (HPV) types 16 and 18 account for approximately 70% of cervical cancer worldwide. Neutralizing HPV prophylactic vaccines offer significant benefit, as they block HPV infection and prevent subsequent disease. However, the three licensed HPV vaccines that cover these two genotypes were produced in eukaryotic cells, which is expensive, particularly for low-income countries where HPV is highest. Here, we report a new HPV16 and -18 bivalent candidate vaccine produced from Escherichia coli. We used two strategies of N-terminal truncation of HPV L1 proteins and soluble non-fusion expression to generate HPV16 and HPV18 L1-only virus-like particles (VLPs) in a scalable process. Through comprehensive characterization of the bivalent candidate vaccine, we confirm lot consistency in a pilot scale-up of 30L, 100L and 500L. Using cryo-EM 3D reconstruction, we found that HPV16 and -18VLPs present in a T=7 icosahedral arrangement, similar in shape and size to that of the native virions. This HPV16/18 bivalent vaccine shares comparable immunogenicity with the licensed vaccines. Overall, we show that the production of a HPV16/18 bivalent vaccine from an E. coli expression system is robust and scalable, with potentially good accessibility worldwide as a population-based immunization strategy.
DateDeposition: Jul 15, 2017 / Header (metadata) release: Sep 6, 2017 / Map release: Sep 6, 2017 / Last update: Sep 6, 2017

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Structure visualization

Movie
  • Surface view with section colored by density value
  • Surface level: 1
  • Imaged by UCSF Chimera
  • Download
  • Surface view colored by radius
  • Surface level: 1
  • Imaged by UCSF Chimera
  • Download
Movie viewer
Structure viewerEM map:
SurfViewMolmilJmol/JSmol
Supplemental images

Downloads & links

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Map

Fileemd_6796.map.gz (map file in CCP4 format, 23329 KB)
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
180 pix
4.44 Å/pix.
= 799.2 Å
180 pix
4.44 Å/pix.
= 799.2 Å
180 pix
4.44 Å/pix.
= 799.2 Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 4.44 Å
Density
Contour Level:1 (by author), 1 (movie #1):
Minimum - Maximum-4.1965246 - 7.800851
Average (Standard dev.)-5.35577E-10 (1)
Details

EMDB XML:

Space Group Number1
Map Geometry
Axis orderXYZ
Dimensions180180180
Origin-92-92-92
Limit878787
Spacing180180180
CellA=B=C: 799.2 Å
α=β=γ: 90 deg.

CCP4 map header:

modeImage stored as Reals
Å/pix. X/Y/Z4.444.444.44
M x/y/z180180180
origin x/y/z0.0000.0000.000
length x/y/z799.200799.200799.200
α/β/γ90.00090.00090.000
start NX/NY/NZ
NX/NY/NZ
MAP C/R/S123
start NC/NR/NS-92-92-92
NC/NR/NS180180180
D min/max/mean-4.1977.801-0.000

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Supplemental data

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Sample components

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Entire Alphapapillomavirus 7

EntireName: Alphapapillomavirus 7 / Number of components: 1

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Component #1: virus, Alphapapillomavirus 7

VirusName: Alphapapillomavirus 7 / Class: VIRUS-LIKE PARTICLE / Empty: Yes / Enveloped: No / Isolate: OTHER
SpeciesSpecies: Alphapapillomavirus 7
Source (engineered)Expression System: Escherichia coli (E. coli)

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Experimental details

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Sample preparation

SpecimenSpecimen state: particle / Method: cryo EM
Sample solutionpH: 7.3
VitrificationCryogen name: ETHANE

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Electron microscopy imaging

Experimental equipment
Model: Tecnai F30 / Image courtesy: FEI Company
ImagingMicroscope: FEI TECNAI F30
Electron gunElectron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Electron dose: 25 e/Å2 / Illumination mode: OTHER
LensImaging mode: BRIGHT FIELD
Specimen HolderModel: OTHER
CameraDetector: FEI FALCON I (4k x 4k)

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Image processing

ProcessingMethod: single particle reconstruction / Number of projections: 700
3D reconstructionResolution: 20 Å / Resolution method: FSC 0.5 CUT-OFF

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