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- EMDB-26736: Hantavirus MAPV Gn(H)/Gc protein in complex with 2 Fabs SNV-24 an... -

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Entry
Database: EMDB / ID: EMD-26736
TitleHantavirus MAPV Gn(H)/Gc protein in complex with 2 Fabs SNV-24 and SNV-53
Map data
Sample
  • Complex: Hanta MAPV virus Gn(H)/Gc protein in complex with 2 Fabs
    • Complex: Hanta MAPV virus Gn(H)/Gc protein
    • Complex: FabFragment antigen-binding
Biological speciesMaporal virus / Homo sapiens (human)
Methodsingle particle reconstruction / negative staining / Resolution: 22.0 Å
AuthorsBinshtein E / Crowe JE
Funding support1 items
OrganizationGrant numberCountry
Defense Advanced Research Projects Agency (DARPA)P3 HR0011-18-2-0001
CitationJournal: Elife / Year: 2023
Title: Antigenic mapping and functional characterization of human New World hantavirus neutralizing antibodies.
Authors: Taylor B Engdahl / Elad Binshtein / Rebecca L Brocato / Natalia A Kuzmina / Lucia M Principe / Steven A Kwilas / Robert K Kim / Nathaniel S Chapman / Monique S Porter / Pablo Guardado-Calvo ...Authors: Taylor B Engdahl / Elad Binshtein / Rebecca L Brocato / Natalia A Kuzmina / Lucia M Principe / Steven A Kwilas / Robert K Kim / Nathaniel S Chapman / Monique S Porter / Pablo Guardado-Calvo / Félix A Rey / Laura S Handal / Summer M Diaz / Irene A Zagol-Ikapitte / Minh H Tran / W Hayes McDonald / Jens Meiler / Joseph X Reidy / Andrew Trivette / Alexander Bukreyev / Jay W Hooper / James E Crowe /
Abstract: Hantaviruses are high-priority emerging pathogens carried by rodents and transmitted to humans by aerosolized excreta or, in rare cases, person-to-person contact. While infections in humans are ...Hantaviruses are high-priority emerging pathogens carried by rodents and transmitted to humans by aerosolized excreta or, in rare cases, person-to-person contact. While infections in humans are relatively rare, mortality rates range from 1 to 40% depending on the hantavirus species. There are currently no FDA-approved vaccines or therapeutics for hantaviruses, and the only treatment for infection is supportive care for respiratory or kidney failure. Additionally, the human humoral immune response to hantavirus infection is incompletely understood, especially the location of major antigenic sites on the viral glycoproteins and conserved neutralizing epitopes. Here, we report antigenic mapping and functional characterization for four neutralizing hantavirus antibodies. The broadly neutralizing antibody SNV-53 targets an interface between Gn/Gc, neutralizes through fusion inhibition and cross-protects against the Old World hantavirus species Hantaan virus when administered pre- or post-exposure. Another broad antibody, SNV-24, also neutralizes through fusion inhibition but targets domain I of Gc and demonstrates weak neutralizing activity to authentic hantaviruses. ANDV-specific, neutralizing antibodies (ANDV-5 and ANDV-34) neutralize through attachment blocking and protect against hantavirus cardiopulmonary syndrome (HCPS) in animals but target two different antigenic faces on the head domain of Gn. Determining the antigenic sites for neutralizing antibodies will contribute to further therapeutic development for hantavirus-related diseases and inform the design of new broadly protective hantavirus vaccines.
History
DepositionApr 25, 2022-
Header (metadata) releaseApr 5, 2023-
Map releaseApr 5, 2023-
UpdateApr 5, 2023-
Current statusApr 5, 2023Processing site: RCSB / Status: Released

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Structure visualization

Supplemental images

emd_26736.png

Downloads & links

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Map

FileDownload / File: emd_26736.map.gz / Format: CCP4 / Size: 3.4 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Voxel sizeX=Y=Z: 4.54 Å
Density
Contour LevelBy AUTHOR: 2.0
Minimum - Maximum-5.0981 - 14.295677
Average (Standard dev.)0.0026393926 (±0.43658802)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions969696
Spacing969696
CellA=B=C: 435.84 Å
α=β=γ: 90.0 °

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Supplemental data

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Half map: #1

Fileemd_26736_half_map_1.map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Half map: #2

Fileemd_26736_half_map_2.map
Projections & Slices
AxesZYX

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Slices (1/2)
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Histogram (log scale)

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Sample components

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Entire : Hanta MAPV virus Gn(H)/Gc protein in complex with 2 Fabs

EntireName: Hanta MAPV virus Gn(H)/Gc protein in complex with 2 Fabs
Components
  • Complex: Hanta MAPV virus Gn(H)/Gc protein in complex with 2 Fabs
    • Complex: Hanta MAPV virus Gn(H)/Gc protein
    • Complex: FabFragment antigen-binding

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Supramolecule #1: Hanta MAPV virus Gn(H)/Gc protein in complex with 2 Fabs

SupramoleculeName: Hanta MAPV virus Gn(H)/Gc protein in complex with 2 Fabs
type: complex / ID: 1 / Chimera: Yes / Parent: 0
Molecular weightTheoretical: 200 KDa

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Supramolecule #2: Hanta MAPV virus Gn(H)/Gc protein

SupramoleculeName: Hanta MAPV virus Gn(H)/Gc protein / type: complex / ID: 2 / Chimera: Yes / Parent: 1
Source (natural)Organism: Maporal virus
Molecular weightTheoretical: 150 KDa

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Supramolecule #3: Fab

SupramoleculeName: Fab / type: complex / ID: 3 / Chimera: Yes / Parent: 1
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 50 KDa

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Experimental details

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Structure determination

Methodnegative staining
Processingsingle particle reconstruction
Aggregation stateparticle

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Sample preparation

Concentration0.01 mg/mL
BufferpH: 8
StainingType: NEGATIVE / Material: uranyl formate
GridModel: Homemade / Material: COPPER / Mesh: 400 / Support film - Material: CARBON / Support film - topology: CONTINUOUS

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Electron microscopy

MicroscopeFEI TECNAI F20
Electron beamAcceleration voltage: 200 kV / Electron source: FIELD EMISSION GUN
Electron opticsC2 aperture diameter: 100.0 µm / Illumination mode: OTHER / Imaging mode: BRIGHT FIELDBright-field microscopy / Cs: 2.2 mm / Nominal defocus max: 1.8 µm / Nominal defocus min: 1.2 µm / Nominal magnification: 50000
Sample stageSpecimen holder model: SIDE ENTRY, EUCENTRIC
Image recordingFilm or detector model: GATAN ULTRASCAN 4000 (4k x 4k) / Detector mode: COUNTING / Number grids imaged: 1 / Number real images: 330 / Average exposure time: 1.0 sec. / Average electron dose: 33.0 e/Å2
Experimental equipment
Model: Tecnai F20 / Image courtesy: FEI Company

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Image processing

Initial angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: cryoSPARC
Final 3D classificationSoftware - Name: cryoSPARC
Final angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: cryoSPARC
Final reconstructionResolution.type: BY AUTHOR / Resolution: 22.0 Å / Resolution method: FSC 0.5 CUT-OFF / Software - Name: cryoSPARC / Number images used: 37800

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Atomic model buiding 1

RefinementSpace: REAL

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