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- EMDB-36441: Cryo-EM structure of dengue virus serotype 3 strain CH53489 spher... -

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Basic information

Entry
Database: EMDB / ID: EMD-36441
TitleCryo-EM structure of dengue virus serotype 3 strain CH53489 spherical particle in complex with human antibody DENV-290 Fab at 37 deg C
Map datapostprocessed map
Sample
  • Complex: Dengue virus serotype 3 strain CH53489 in complex with human antibody DENV-290 Fab at 37 deg C
    • Complex: Dengue virus serotype 3 strain CH53489
      • Protein or peptide: Envelope protein
      • Protein or peptide: Membrane protein
    • Complex: Human antibody DENV-290 Fab (heavy and light chain)
      • Protein or peptide: Human antibody DENV-290 heavy chain
      • Protein or peptide: Human antibody DENV-290 light chain
Keywordsdengue virus / human antibody / dengue-antibody structure / virus
Biological speciesDengue virus type 3 / Homo sapiens (human) / Aedes albopictus (Asian tiger mosquito)
Methodsingle particle reconstruction / cryo EM / Resolution: 5.6 Å
AuthorsFibriansah G / Ng TS / Tan AWK / Shi J / Lok SM
Funding support Singapore, 1 items
OrganizationGrant numberCountry
National Research Foundation (NRF, Singapore)NRF-NRFI2016-01 Singapore
CitationJournal: To Be Published
Title: Ultrapotent human antibodies lock E protein dimers of diverse DENV3 morphological variants
Authors: Fibriansah G / Ng TS / Lim XN / Shebanova A / Ng LC / Tan SL / Tan AWK / Shi J / Crowe JE / Lok SM
History
DepositionJun 6, 2023-
Header (metadata) releaseJun 12, 2024-
Map releaseJun 12, 2024-
UpdateJun 12, 2024-
Current statusJun 12, 2024Processing site: PDBj / Status: Released

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Structure visualization

Supplemental images

emd_36441.png

Downloads & links

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Map

FileDownload / File: emd_36441.map.gz / Format: CCP4 / Size: 512 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Annotationpostprocessed map
Voxel sizeX=Y=Z: 1.71 Å
Density
Contour LevelBy AUTHOR: 2.0
Minimum - Maximum-5.6456323 - 13.366391
Average (Standard dev.)-0.000000009142066 (±1.0)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin-256-256-256
Dimensions512512512
Spacing512512512
CellA=B=C: 875.52 Å
α=β=γ: 90.0 °

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Supplemental data

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Mask #1

Fileemd_36441_msk_1.map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Half map: half map 1

Fileemd_36441_half_map_1.map
Annotationhalf map 1
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Half map: half map 2

Fileemd_36441_half_map_2.map
Annotationhalf map 2
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Sample components

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Entire : Dengue virus serotype 3 strain CH53489 in complex with human anti...

EntireName: Dengue virus serotype 3 strain CH53489 in complex with human antibody DENV-290 Fab at 37 deg C
Components
  • Complex: Dengue virus serotype 3 strain CH53489 in complex with human antibody DENV-290 Fab at 37 deg C
    • Complex: Dengue virus serotype 3 strain CH53489
      • Protein or peptide: Envelope protein
      • Protein or peptide: Membrane protein
    • Complex: Human antibody DENV-290 Fab (heavy and light chain)
      • Protein or peptide: Human antibody DENV-290 heavy chain
      • Protein or peptide: Human antibody DENV-290 light chain

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Supramolecule #1: Dengue virus serotype 3 strain CH53489 in complex with human anti...

SupramoleculeName: Dengue virus serotype 3 strain CH53489 in complex with human antibody DENV-290 Fab at 37 deg C
type: complex / ID: 1 / Parent: 0 / Macromolecule list: all

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Supramolecule #2: Dengue virus serotype 3 strain CH53489

SupramoleculeName: Dengue virus serotype 3 strain CH53489 / type: complex / ID: 2 / Parent: 1 / Macromolecule list: #1-#2
Source (natural)Organism: Dengue virus type 3

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Supramolecule #3: Human antibody DENV-290 Fab (heavy and light chain)

SupramoleculeName: Human antibody DENV-290 Fab (heavy and light chain) / type: complex / ID: 3 / Parent: 1 / Macromolecule list: #3-#4
Source (natural)Organism: Homo sapiens (human)

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Macromolecule #1: Envelope protein

MacromoleculeName: Envelope protein / type: protein_or_peptide / ID: 1 / Enantiomer: LEVO
Source (natural)Organism: Aedes albopictus (Asian tiger mosquito)
SequenceString: mrcvgvgnrd fveglsgat w vdvvlehg gc vttmakn kpt ldielq ktea tqlat lrklc iegk itnitt dsr cptqgea vl peeqdqny v ckhtyvdrg wgngcglfgk gslvtcakf q clepiegk vv qyenlky tvi itvhtg dqhq vgnet ...String:
mrcvgvgnrd fveglsgat w vdvvlehg gc vttmakn kpt ldielq ktea tqlat lrklc iegk itnitt dsr cptqgea vl peeqdqny v ckhtyvdrg wgngcglfgk gslvtcakf q clepiegk vv qyenlky tvi itvhtg dqhq vgnet qgvta eitp qastte ail peygtlg le csprtgld f nemilltmk nkawmvhrqw ffdlplpwa s gattetpt wn rkellvt fkn ahakkq evvv lgsqe gamht altg ateiqn sgg tsifagh lk crlkmdkl e lkgmsyamc tntfvlkkev setqhgtil i kveykged ap ckipfst edg qgkahn grli tanpv vtkke epvn ieaepp fge snivigi gd nalkinwy k kgssigkmf eatargarrm ailgdtawd f gsvggvln sl gkmvhqi fgs aytalf sgvs wvmki gigvl ltwi glnskn tsm sfsciai gi itlylgav v qa

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Macromolecule #2: Membrane protein

MacromoleculeName: Membrane protein / type: protein_or_peptide / ID: 2 / Enantiomer: LEVO
Source (natural)Organism: Aedes albopictus (Asian tiger mosquito)
SequenceString:
svala phvg mgldtr tqt wmsaega wr qvekvetw a lrhpgftil alflahyigt sltqkvvif i llmlvtps mt

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Macromolecule #3: Human antibody DENV-290 heavy chain

MacromoleculeName: Human antibody DENV-290 heavy chain / type: protein_or_peptide / ID: 3 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString:
QVELVESGGD VVQPGKSLRL SCAASGFTFT NYAMHWLRQA PGKGLEWVAV ISSDVNDKYY ADSVKGRFTI SRDNSKNTLY LQMNSLTPED TAVYYCAREQ AVGTNPWAFD YWGQGTLVTV SS

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Macromolecule #4: Human antibody DENV-290 light chain

MacromoleculeName: Human antibody DENV-290 light chain / type: protein_or_peptide / ID: 4 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString:
HIVMTQSPLS LSVTPGQPAS ISCKSSQISS WGSDGKTYLY WYLQKPGQSP QLLIYEVSSR FSGVSDRFSG SGSGTDFTLK ISRVQAEDVG LYYCMQGLHL PLTFGQGTRL EIK

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Experimental details

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Structure determination

Methodcryo EM
Processingsingle particle reconstruction
Aggregation stateparticle

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Sample preparation

BufferpH: 8
Details: NTE buffer (12 mM Tris-HCl pH 8.0, 120 mM NaCl and 1 mM EDTA)
VitrificationCryogen name: ETHANE
DetailsThe purified virus was mixed with DENV-290 Fab at a molar ratio of 1.5 Fab for every E-protein and then the mixture was incubated at 4 deg C for 30 min. The complex was further incubated at 37 deg C for 30 min, and then incubated at 4 deg C for another 2 h

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Image recordingFilm or detector model: FEI FALCON II (4k x 4k) / Average electron dose: 18.0 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 5.0 µm / Nominal defocus min: 0.5 µm
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Startup modelType of model: EMDB MAP
EMDB ID:

5933

Final reconstructionApplied symmetry - Point group: I (icosahedral) / Resolution.type: BY AUTHOR / Resolution: 5.6 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: RELION / Number images used: 23584
Initial angle assignmentType: MAXIMUM LIKELIHOOD
Final angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: RELION

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