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- EMDB-46044: Cryo-EM structure of DA03E17 Fab in complex with influenza virus ... -

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Basic information

Entry
Database: EMDB / ID: EMD-46044
TitleCryo-EM structure of DA03E17 Fab in complex with influenza virus neuraminidase from B/Colorado/06/2017
Map data
Sample
  • Complex: DA03E17 Fab in complex with influenza virus neuraminidase from B/Colorado/06/2017
    • Protein or peptide: DA03E17 Fab heavy chain
    • Protein or peptide: DA03E17 Fab light chain
    • Protein or peptide: Neuraminidase
  • Ligand: CALCIUM ION
KeywordsNeuraminidase / Hydrolase / Antibody complex / VIRAL PROTEIN / VIRAL PROTEIN-IMMUNE SYSTEM complex
Function / homology
Function and homology information


exo-alpha-sialidase / exo-alpha-sialidase activity / carbohydrate metabolic process / host cell plasma membrane / virion membrane / metal ion binding / membrane
Similarity search - Function
Glycoside hydrolase, family 34 / Neuraminidase / Sialidase superfamily
Similarity search - Domain/homology
Biological speciesHomo sapiens (human) / Influenza B virus (B/Colorado/06/2017)
Methodsingle particle reconstruction / cryo EM / Resolution: 2.47 Å
AuthorsJo G / Ward AB
Funding support United States, 1 items
OrganizationGrant numberCountry
National Institutes of Health/National Institute Of Allergy and Infectious Diseases (NIH/NIAID)75N93019C00051 United States
CitationJournal: Nat Commun / Year: 2025
Title: Structural basis of broad protection against influenza virus by human antibodies targeting the neuraminidase active site via a recurring motif in CDR H3.
Authors: Gyunghee Jo / Seiya Yamayoshi / Krystal M Ma / Olivia Swanson / Jonathan L Torres / James A Ferguson / Monica L Fernández-Quintero / Jiachen Huang / Jeffrey Copps / Alesandra J Rodriguez / ...Authors: Gyunghee Jo / Seiya Yamayoshi / Krystal M Ma / Olivia Swanson / Jonathan L Torres / James A Ferguson / Monica L Fernández-Quintero / Jiachen Huang / Jeffrey Copps / Alesandra J Rodriguez / Jon M Steichen / Yoshihiro Kawaoka / Julianna Han / Andrew B Ward /
Abstract: Influenza viruses evolve rapidly, driving seasonal epidemics and posing global pandemic threats. While neuraminidase (NA) has emerged as a vaccine target, shared molecular features of NA antibody ...Influenza viruses evolve rapidly, driving seasonal epidemics and posing global pandemic threats. While neuraminidase (NA) has emerged as a vaccine target, shared molecular features of NA antibody responses are still not well understood. Here, we describe cryo-electron microscopy structures of the broadly protective human antibody DA03E17, which was previously identified from an H1N1-infected donor, in complex with NA from A/H1N1, A/H3N2, and B/Victoria-lineage viruses. DA03E17 targets the highly conserved NA active site using its long CDR H3, which features a DR (Asp-Arg) motif that engages catalytic residues and mimics sialic acid interactions. We further demonstrate that this motif is conserved among several NA active site-targeting antibodies, indicating a common receptor mimicry strategy. We also identified BCR sequences containing this DR motif across all donors in a healthy human repertoire database, suggesting that such precursors may be relatively common and have vaccine targeting potential. Our findings reveal shared molecular features in NA active site-targeting antibodies that can be harnessed to design broad, immune-focused influenza vaccines.
History
DepositionAug 2, 2024-
Header (metadata) releaseAug 13, 2025-
Map releaseAug 13, 2025-
UpdateAug 13, 2025-
Current statusAug 13, 2025Processing site: RCSB / Status: Released

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Structure visualization

Supplemental images

emd_46044.png

Downloads & links

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Map

FileDownload / File: emd_46044.map.gz / Format: CCP4 / Size: 512 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
0.73 Å/pix.
x 512 pix.
= 371.2 Å		0.73 Å/pix.
x 512 pix.
= 371.2 Å		0.73 Å/pix.
x 512 pix.
= 371.2 Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 0.725 Å
Density

Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 0.08
Minimum - Maximum-0.22318101 - 0.431287
Average (Standard dev.)-0.00004409872 (±0.008485948)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions512512512
Spacing512512512
CellA=B=C: 371.2 Å
α=β=γ: 90.0 °

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Supplemental data

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Mask #1

Fileemd_46044_msk_1.map
Projections & Slices
AxesZYX

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Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Half map: #2

Fileemd_46044_half_map_1.map
Projections & Slices
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Histogram

Histogram (log scale)

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Half map: #1

Fileemd_46044_half_map_2.map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
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Histogram

Histogram (log scale)

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Sample components

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Entire : DA03E17 Fab in complex with influenza virus neuraminidase from B/...

EntireName: DA03E17 Fab in complex with influenza virus neuraminidase from B/Colorado/06/2017
Components
  • Complex: DA03E17 Fab in complex with influenza virus neuraminidase from B/Colorado/06/2017
    • Protein or peptide: DA03E17 Fab heavy chain
    • Protein or peptide: DA03E17 Fab light chain
    • Protein or peptide: Neuraminidase
  • Ligand: CALCIUM ION

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Supramolecule #1: DA03E17 Fab in complex with influenza virus neuraminidase from B/...

SupramoleculeName: DA03E17 Fab in complex with influenza virus neuraminidase from B/Colorado/06/2017
type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#3
Source (natural)Organism: Homo sapiens (human)

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Macromolecule #1: DA03E17 Fab heavy chain

MacromoleculeName: DA03E17 Fab heavy chain / type: protein_or_peptide / ID: 1 / Number of copies: 4 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 24.554438 KDa
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString: EVQLVESGPG LVKPSQTLSL TCTVSGGSFS SGGYLWSWVR QHPGKGLEWI GYILYSGSPY YNPSLESRAT ISLDTSKNQF SLRLISVTA ADAAMYYCAR VDGSGNTDRY YFYGMDVWGQ GTMVTVSSAS TKGPSVFPLA PSSKSTSGGT AALGCLVKDY F PEPVTVSW ...String:
EVQLVESGPG LVKPSQTLSL TCTVSGGSFS SGGYLWSWVR QHPGKGLEWI GYILYSGSPY YNPSLESRAT ISLDTSKNQF SLRLISVTA ADAAMYYCAR VDGSGNTDRY YFYGMDVWGQ GTMVTVSSAS TKGPSVFPLA PSSKSTSGGT AALGCLVKDY F PEPVTVSW NSGALTSGVH TFPAVLQSSG LYSLSSVVTV PSSSLGTQTY ICNVNHKPSN TKVDKRVEPK SCD

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Macromolecule #2: DA03E17 Fab light chain

MacromoleculeName: DA03E17 Fab light chain / type: protein_or_peptide / ID: 2 / Number of copies: 4 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 23.486031 KDa
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString: DIQMTQSPSS VSASVGDRVT ITCRASRGIG DWLAWYQQKP GKAPKLLIYA ASSLQRGVPS RFSGSGSGTD FTLTISSLQP DDFATYYCQ QADGWEVWTF GQGTKVDVKR TVAAPSVFIF PPSDEQLKSG TASVVCLLNN FYPREAKVQW KVDNALQSGN S QESVTEQD ...String:
DIQMTQSPSS VSASVGDRVT ITCRASRGIG DWLAWYQQKP GKAPKLLIYA ASSLQRGVPS RFSGSGSGTD FTLTISSLQP DDFATYYCQ QADGWEVWTF GQGTKVDVKR TVAAPSVFIF PPSDEQLKSG TASVVCLLNN FYPREAKVQW KVDNALQSGN S QESVTEQD SKDSTYSLSS TLTLSKADYE KHKVYACEVT HQGLSSPVTK SFNRGEC

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Macromolecule #3: Neuraminidase

MacromoleculeName: Neuraminidase / type: protein_or_peptide / ID: 3 / Number of copies: 4 / Enantiomer: LEVO / EC number: exo-alpha-sialidase
Source (natural)Organism: Influenza B virus (B/Colorado/06/2017)
Molecular weightTheoretical: 53.536656 KDa
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString: MYSMQLASCV TLTLVLLVNS QHHHHHHGSA WSHPQFEKGG SSSDYSDLQR VKQELLEEVK KELQKVKEEI IEAFVQELRK RGSLVPRGS GGPEPEWTYP RLSCPGSTFQ KALLISPHRF GETKGNSAPL IIREPFVACG PNECKHFALT HYAAQPGGYY N GTRGDRNK ...String:
MYSMQLASCV TLTLVLLVNS QHHHHHHGSA WSHPQFEKGG SSSDYSDLQR VKQELLEEVK KELQKVKEEI IEAFVQELRK RGSLVPRGS GGPEPEWTYP RLSCPGSTFQ KALLISPHRF GETKGNSAPL IIREPFVACG PNECKHFALT HYAAQPGGYY N GTRGDRNK LRHLISVKLG KIPTVENSIF HMAAWSGSAC HDGKEWTYIG VDGPDNNALL KVKYGEAYTD TYHSYANNIL RT QESACNC IGGNCYLMIT DGSASGVSEC RFLKIREGRI IKEIFPTGRV KHTEECTCGF ASNKTIECAC RDNRYTAKRP FVK LNVETD TAEIRLMCTD TYLDTPRPND GSITGPCESD GDKGSGGIKG GFVHQRMKSK IGRWYSRTMS QTERMGMGLY VKYG GDPWA DSDALAFSGV MVSMKEPGWY SFGFEIKDKK CDVPCIGIEM VHDGGKETWH SAATAIYCLM GSGQLLWDTV TGVDM AL

UniProtKB: Neuraminidase

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Macromolecule #5: CALCIUM ION

MacromoleculeName: CALCIUM ION / type: ligand / ID: 5 / Number of copies: 4 / Formula: CA
Molecular weightTheoretical: 40.078 Da

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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

Concentration0.4 mg/mL
BufferpH: 7.4
VitrificationCryogen name: ETHANE / Instrument: FEI VITROBOT MARK IV

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Electron microscopy

MicroscopeTFS GLACIOS
Image recordingFilm or detector model: FEI FALCON IV (4k x 4k) / Average electron dose: 45.06 e/Å2
Electron beamAcceleration voltage: 200 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 1.4000000000000001 µm / Nominal defocus min: 0.7000000000000001 µm / Nominal magnification: 190000
Sample stageSpecimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER

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Image processing

CTF correctionSoftware - Name: cryoSPARC (ver. 4.2.1) / Type: PHASE FLIPPING AND AMPLITUDE CORRECTION
Startup modelType of model: INSILICO MODEL / Details: cryoSPARC - ab initio
Final reconstructionApplied symmetry - Point group: C4 (4 fold cyclic) / Resolution.type: BY AUTHOR / Resolution: 2.47 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: cryoSPARC (ver. 4.2.1) / Number images used: 420785
Initial angle assignmentType: MAXIMUM LIKELIHOOD
Final angle assignmentType: MAXIMUM LIKELIHOOD
FSC plot (resolution estimation)

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