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- EMDB-64401: Cryo-EM structure of the N-terminal domain of Omicron BA.1 in com... -

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Entry
Database: EMDB / ID: EMD-64401
TitleCryo-EM structure of the N-terminal domain of Omicron BA.1 in complex with nanobody N103 and S2L20 Fab
Map data
Sample
  • Complex: the N-terminal domain of Omicron BA.1 in complex with nanobody N103 and S2L20 Fab
    • Protein or peptide: S2L20 light chain
    • Protein or peptide: S2L20 heavy chain
    • Protein or peptide: nanobody N103
    • Protein or peptide: Spike glycoprotein
  • Ligand: 2-acetamido-2-deoxy-beta-D-glucopyranose
Keywordscomplex / VIRAL PROTEIN/IMMUNE SYSTEM / VIRAL PROTEIN-IMMUNE SYSTEM complex / VIRAL PROTEIN
Function / homology
Function and homology information


symbiont-mediated disruption of host tissue / Maturation of spike protein / Translation of Structural Proteins / Virion Assembly and Release / host cell surface / host extracellular region / symbiont-mediated-mediated suppression of host tetherin activity / Induction of Cell-Cell Fusion / structural constituent of virion / positive regulation of viral entry into host cell ...symbiont-mediated disruption of host tissue / Maturation of spike protein / Translation of Structural Proteins / Virion Assembly and Release / host cell surface / host extracellular region / symbiont-mediated-mediated suppression of host tetherin activity / Induction of Cell-Cell Fusion / structural constituent of virion / positive regulation of viral entry into host cell / membrane fusion / host cell endoplasmic reticulum-Golgi intermediate compartment membrane / Attachment and Entry / entry receptor-mediated virion attachment to host cell / receptor-mediated virion attachment to host cell / host cell surface receptor binding / symbiont-mediated suppression of host innate immune response / endocytosis involved in viral entry into host cell / receptor ligand activity / fusion of virus membrane with host plasma membrane / fusion of virus membrane with host endosome membrane / viral envelope / symbiont entry into host cell / virion attachment to host cell / host cell plasma membrane / SARS-CoV-2 activates/modulates innate and adaptive immune responses / virion membrane / membrane / identical protein binding / plasma membrane
Similarity search - Function
Spike (S) protein S1 subunit, receptor-binding domain, SARS-CoV-2 / Spike (S) protein S1 subunit, N-terminal domain, SARS-CoV-like / Coronavirus spike glycoprotein S1, C-terminal / Coronavirus spike glycoprotein S1, C-terminal / Spike glycoprotein, N-terminal domain superfamily / Spike S1 subunit, receptor binding domain superfamily, betacoronavirus / Spike glycoprotein, betacoronavirus / Betacoronavirus spike (S) glycoprotein S1 subunit N-terminal (NTD) domain profile. / Spike glycoprotein S1, N-terminal domain, betacoronavirus-like / Betacoronavirus-like spike glycoprotein S1, N-terminal ...Spike (S) protein S1 subunit, receptor-binding domain, SARS-CoV-2 / Spike (S) protein S1 subunit, N-terminal domain, SARS-CoV-like / Coronavirus spike glycoprotein S1, C-terminal / Coronavirus spike glycoprotein S1, C-terminal / Spike glycoprotein, N-terminal domain superfamily / Spike S1 subunit, receptor binding domain superfamily, betacoronavirus / Spike glycoprotein, betacoronavirus / Betacoronavirus spike (S) glycoprotein S1 subunit N-terminal (NTD) domain profile. / Spike glycoprotein S1, N-terminal domain, betacoronavirus-like / Betacoronavirus-like spike glycoprotein S1, N-terminal / Betacoronavirus spike (S) glycoprotein S1 subunit C-terminal (CTD) domain profile. / Spike (S) protein S1 subunit, receptor-binding domain, betacoronavirus / Betacoronavirus spike glycoprotein S1, receptor binding / Spike glycoprotein S2 superfamily, coronavirus / Spike glycoprotein S2, coronavirus, heptad repeat 1 / Spike glycoprotein S2, coronavirus, heptad repeat 2 / Coronavirus spike (S) glycoprotein S2 subunit heptad repeat 1 (HR1) region profile. / Coronavirus spike (S) glycoprotein S2 subunit heptad repeat 2 (HR2) region profile. / Spike glycoprotein S2, coronavirus / Coronavirus spike glycoprotein S2
Similarity search - Domain/homology
Biological speciesVicugna pacos (alpaca) / Homo sapiens (human) / Severe acute respiratory syndrome coronavirus 2
Methodsingle particle reconstruction / cryo EM / Resolution: 2.77 Å
AuthorsLiu B / Liu HH / Wang CM / Han P / Wang QH
Funding support China, 1 items
OrganizationGrant numberCountry
National Natural Science Foundation of China (NSFC) China
CitationJournal: Mol Ther / Year: 2026
Title: From weak but broad to potent and universal: A trispecific antibody against conserved SARS-CoV-2 spike epitopes.
Authors: Xiaodong Tian / Chunmei Wang / Pu Han / Honghui Liu / Lili Wu / Xiaoyun Wang / Shufan Yu / Yanning Zhang / Binbin Zhao / Jiangning Liu / George Fu Gao / Qihui Wang /
Abstract: Most neutralizing antibodies against severe acute respiratory syndrome coronavirus 2 target the receptor-binding domain (RBD). However, due to high immune pressure, the RBD accumulates mutations, ...Most neutralizing antibodies against severe acute respiratory syndrome coronavirus 2 target the receptor-binding domain (RBD). However, due to high immune pressure, the RBD accumulates mutations, thus significantly reducing antibody efficacy against emerging variants/subvariants. Although some RBD-targeting antibodies bind conserved epitopes, they usually exhibit weak-to-moderate neutralization. Similarly, antibodies against the N-terminal domain (NTD) or S2 subunit often retain broad binding but generally lack potent neutralization. To address this, we initially identified the broadly reactive nanobody, N103, which exhibited weak neutralizing potency. Structural and functional analyses revealed that N103 targets a conserved NTD epitope and triggers S1 subunit shedding, thereby destabilizing the spike trimer through an allosteric mechanism. Leveraging this insight, we engineered a trispecific antibody combining N103 with antibodies targeting the conserved RBD and S2 epitopes. This design synergistically integrated their distinct binding profiles and mechanisms, achieving potent and broad neutralization against both pseudoviruses and authentic viruses, including the immune-evasive BA.2.86 subvariant. Furthermore, challenge studies in human angiotensin-converting enzyme 2 knockin mice demonstrated robust in vivo protection. Our findings highlight a cooperative multi-target strategy in which antibodies with limited individual potency can collectively achieve broad and potent neutralization through rational design. This approach provides a promising framework for next-generation antibody therapeutics.
History
DepositionApr 28, 2025-
Header (metadata) releaseMay 6, 2026-
Map releaseMay 6, 2026-
UpdateJun 10, 2026-
Current statusJun 10, 2026Processing site: PDBc / Status: Released

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Structure visualization

Supplemental images

emd_64401.png

Downloads & links

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Map

FileDownload / File: emd_64401.map.gz / Format: CCP4 / Size: 244.1 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
0.69 Å/pix.
x 400 pix.
= 276. Å		0.69 Å/pix.
x 400 pix.
= 276. Å		0.69 Å/pix.
x 400 pix.
= 276. Å

Surface

Projections

Slices (1/3)

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Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 0.69 Å
Density

Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 0.21
Minimum - Maximum-0.37069067 - 1.0029796
Average (Standard dev.)-0.0017537769 (±0.021396488)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions400400400
Spacing400400400
CellA=B=C: 276.0 Å
α=β=γ: 90.0 °

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Supplemental data

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Half map: #2

Fileemd_64401_half_map_1.map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Half map: #1

Fileemd_64401_half_map_2.map
Projections & Slices
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Histogram (log scale)

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Sample components

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Entire : the N-terminal domain of Omicron BA.1 in complex with nanobody N1...

EntireName: the N-terminal domain of Omicron BA.1 in complex with nanobody N103 and S2L20 Fab
Components
  • Complex: the N-terminal domain of Omicron BA.1 in complex with nanobody N103 and S2L20 Fab
    • Protein or peptide: S2L20 light chain
    • Protein or peptide: S2L20 heavy chain
    • Protein or peptide: nanobody N103
    • Protein or peptide: Spike glycoprotein
  • Ligand: 2-acetamido-2-deoxy-beta-D-glucopyranose

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Supramolecule #1: the N-terminal domain of Omicron BA.1 in complex with nanobody N1...

SupramoleculeName: the N-terminal domain of Omicron BA.1 in complex with nanobody N103 and S2L20 Fab
type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#4
Source (natural)Organism: Vicugna pacos (alpaca)

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Macromolecule #1: S2L20 light chain

MacromoleculeName: S2L20 light chain / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 26.017973 KDa
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString: METDTLLLWV LLLWVPGSTG DVIWMTQSPS SLSASVGDRV TITCQASQDI RFYLNWYQQK PGKAPKLLIS DASNMETGVP SRFSGSGSG TDFTFTISSL QPEDIATYYC QQYDNLPFTF GPGTKVDFKR TVAAPSVFIF PPSDEQLKSG TASVVCLLNN F YPREAKVQ ...String:
METDTLLLWV LLLWVPGSTG DVIWMTQSPS SLSASVGDRV TITCQASQDI RFYLNWYQQK PGKAPKLLIS DASNMETGVP SRFSGSGSG TDFTFTISSL QPEDIATYYC QQYDNLPFTF GPGTKVDFKR TVAAPSVFIF PPSDEQLKSG TASVVCLLNN F YPREAKVQ WKVDNALQSG NSQESVTEQD SKDSTYSLSS TLTLSKADYE KHKVYACEVT HQGLSSPVTK SFNRGECS

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Macromolecule #2: S2L20 heavy chain

MacromoleculeName: S2L20 heavy chain / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 52.041504 KDa
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString: METDTLLLWV LLLWVPGSTG DEVQLVESGG GVVQPGGSLR LSCAASGFTF NSYGMHWVRQ APGKGLEWVA FIRYDGGNKY YADSVKGRF TISRDNSKNT LYLQMKSLRA EDTAVYYCAN LKDSRYSGSY YDYWGQGTLV TVSSASTKGP SVFPLAPSSK S TSGGTAAL ...String:
METDTLLLWV LLLWVPGSTG DEVQLVESGG GVVQPGGSLR LSCAASGFTF NSYGMHWVRQ APGKGLEWVA FIRYDGGNKY YADSVKGRF TISRDNSKNT LYLQMKSLRA EDTAVYYCAN LKDSRYSGSY YDYWGQGTLV TVSSASTKGP SVFPLAPSSK S TSGGTAAL GCLVKDYFPE PVTVSWNSGA LTSGVHTFPA VLQSSGLYSL SSVVTVPSSS LGTQTYICNV NHKPSNTKVD KR VEPKSCD KTHTCPPCPA PELLGGPSVF LFPPKPKDTL MISRTPEVTC VVVDVSHEDP EVKFNWYVDG VEVHNAKTKP REE QYNSTY RVVSVLTVLH QDWLNGKEYK CKVSNKALPA PIEKTISKAK GQPREPQVYT LPPSRDELTK NQVSLTCLVK GFYP SDIAV EWESNGQPEN NYKTTPPVLD SDGSFFLYSK LTVDKSRWQQ GNVFSCSVMH EALHNHYTQK SLSLSPGK

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Macromolecule #3: nanobody N103

MacromoleculeName: nanobody N103 / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Vicugna pacos (alpaca)
Molecular weightTheoretical: 12.959353 KDa
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString:
QVQLQESGGG LVQPGGSLRL SCAASVNLFI GATMAWYRQA PGNQRELVAT IGAGGTTNYA DSVKGRFTAS RDDAKKTIYL QMNSLTPED TAVYYCNYGS MSNPRTSGPN AFWGRGTQVT VSS

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Macromolecule #4: Spike glycoprotein

MacromoleculeName: Spike glycoprotein / type: protein_or_peptide / ID: 4 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Severe acute respiratory syndrome coronavirus 2 / Strain: Omicron/BA.1
Molecular weightTheoretical: 34.909473 KDa
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString: MFVFLVLLPL VSSQCVNLTT RTQLPPAYTN SFTRGVYYPD KVFRSSVLHS TQDLFLPFFS NVTWFHVISG TNGTKRFDNP VLPFNDGVY FASIEKSNII RGWIFGTTLD SKTQSLLIVN NATNVVIKVC EFQFCNDPFL DHKNNKSWME SEFRVYSSAN N CTFEYVSQ ...String:
MFVFLVLLPL VSSQCVNLTT RTQLPPAYTN SFTRGVYYPD KVFRSSVLHS TQDLFLPFFS NVTWFHVISG TNGTKRFDNP VLPFNDGVY FASIEKSNII RGWIFGTTLD SKTQSLLIVN NATNVVIKVC EFQFCNDPFL DHKNNKSWME SEFRVYSSAN N CTFEYVSQ PFLMDLEGKQ GNFKNLREFV FKNIDGYFKI YSKHTPIREP EDLPQGFSAL EPLVDLPIGI NITRFQTLLA LH RSYLTPG DSSSGWTAGA AAYYVGYLQP RTFLLKYNEN GTITDAVDCA LDPLSETKCT LKSHHHHHH

UniProtKB: Spike glycoprotein

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Macromolecule #6: 2-acetamido-2-deoxy-beta-D-glucopyranose

MacromoleculeName: 2-acetamido-2-deoxy-beta-D-glucopyranose / type: ligand / ID: 6 / Number of copies: 4 / Formula: NAG
Molecular weightTheoretical: 221.208 Da
Chemical component information

ChemComp-NAG:
2-acetamido-2-deoxy-beta-D-glucopyranose

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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
VitrificationCryogen name: ETHANE

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Electron microscopy

MicroscopeFEI TALOS ARCTICA
Image recordingFilm or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Average electron dose: 60.0 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 2.0 µm / Nominal defocus min: 1.0 µm
Experimental equipment
Model: Talos Arctica / Image courtesy: FEI Company

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Image processing

CTF correctionType: PHASE FLIPPING AND AMPLITUDE CORRECTION
Startup modelType of model: INSILICO MODEL
Final reconstructionResolution.type: BY AUTHOR / Resolution: 2.77 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 185260
Initial angle assignmentType: MAXIMUM LIKELIHOOD
Final angle assignmentType: MAXIMUM LIKELIHOOD

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