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- EMDB-27723: Cryo-EM Structure of Antibody SKW11 in complex with Western Equin... -

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Basic information

Entry
Database: EMDB / ID: EMD-27723
TitleCryo-EM Structure of Antibody SKW11 in complex with Western Equine Encephalitis Virus spike (local refinement from VLP particles)
Map data
Sample
  • Complex: Western Equine Encephalitis Virus trimer in complex with Antibody Fab SKW11
    • Protein or peptide: Antibody SKW11 heavy chain
    • Protein or peptide: Antibody SKW11 light chain
    • Protein or peptide: Spike glycoprotein E1
    • Protein or peptide: Spike glycoprotein E2
  • Ligand: 2-acetamido-2-deoxy-beta-D-glucopyranose
KeywordsAntibody / SKW11 / Western Equine Encephalitis Virus / broadly neutralizing / IMMUNE SYSTEM
Function / homology
Function and homology information


togavirin / T=4 icosahedral viral capsid / symbiont-mediated suppression of host toll-like receptor signaling pathway / host cell cytoplasm / symbiont entry into host cell / symbiont-mediated suppression of host gene expression / serine-type endopeptidase activity / fusion of virus membrane with host endosome membrane / host cell nucleus / virion attachment to host cell ...togavirin / T=4 icosahedral viral capsid / symbiont-mediated suppression of host toll-like receptor signaling pathway / host cell cytoplasm / symbiont entry into host cell / symbiont-mediated suppression of host gene expression / serine-type endopeptidase activity / fusion of virus membrane with host endosome membrane / host cell nucleus / virion attachment to host cell / host cell plasma membrane / virion membrane / structural molecule activity / proteolysis / RNA binding / membrane
Similarity search - Function
Alphavirus E2 glycoprotein, domain B / Peptidase S3, togavirin / Alphavirus E2 glycoprotein / Alphavirus E3 spike glycoprotein / Alphavirus E1 glycoprotein / Alphavirus E2 glycoprotein, domain A / Alphavirus E2 glycoprotein, domain C / Alphavirus E2 glycoprotein / Alphavirus core protein / Alphavirus E3 glycoprotein ...Alphavirus E2 glycoprotein, domain B / Peptidase S3, togavirin / Alphavirus E2 glycoprotein / Alphavirus E3 spike glycoprotein / Alphavirus E1 glycoprotein / Alphavirus E2 glycoprotein, domain A / Alphavirus E2 glycoprotein, domain C / Alphavirus E2 glycoprotein / Alphavirus core protein / Alphavirus E3 glycoprotein / Alphavirus E1 glycoprotein / Alphavirus core protein (CP) domain profile. / Flavivirus/Alphavirus glycoprotein, immunoglobulin-like domain superfamily / Flavivirus glycoprotein, central and dimerisation domain superfamily / Flaviviral glycoprotein E, dimerisation domain / Immunoglobulin E-set / Peptidase S1, PA clan, chymotrypsin-like fold / Peptidase S1, PA clan
Similarity search - Domain/homology
Structural polyprotein
Similarity search - Component
Biological speciesWestern equine encephalitis virus / Macaca fascicularis (crab-eating macaque)
Methodsingle particle reconstruction / cryo EM / Resolution: 5.84 Å
AuthorsCerutti G / Verardi R / Roederer M / Shapiro L
Funding support United States, 1 items
OrganizationGrant numberCountry
National Institutes of Health/National Institute Of Allergy and Infectious Diseases (NIH/NIAID) United States
CitationJournal: Cell / Year: 2023
Title: Vaccine elicitation and structural basis for antibody protection against alphaviruses.
Authors: Matthew S Sutton / Sergei Pletnev / Victoria Callahan / Sungyoul Ko / Yaroslav Tsybovsky / Tatsiana Bylund / Ryan G Casner / Gabriele Cerutti / Christina L Gardner / Veronica Guirguis / ...Authors: Matthew S Sutton / Sergei Pletnev / Victoria Callahan / Sungyoul Ko / Yaroslav Tsybovsky / Tatsiana Bylund / Ryan G Casner / Gabriele Cerutti / Christina L Gardner / Veronica Guirguis / Raffaello Verardi / Baoshan Zhang / David Ambrozak / Margaret Beddall / Hong Lei / Eun Sung Yang / Tracy Liu / Amy R Henry / Reda Rawi / Arne Schön / Chaim A Schramm / Chen-Hsiang Shen / Wei Shi / Tyler Stephens / Yongping Yang / Maria Burgos Florez / Julie E Ledgerwood / Crystal W Burke / Lawrence Shapiro / Julie M Fox / Peter D Kwong / Mario Roederer /
Abstract: Alphaviruses are RNA viruses that represent emerging public health threats. To identify protective antibodies, we immunized macaques with a mixture of western, eastern, and Venezuelan equine ...Alphaviruses are RNA viruses that represent emerging public health threats. To identify protective antibodies, we immunized macaques with a mixture of western, eastern, and Venezuelan equine encephalitis virus-like particles (VLPs), a regimen that protects against aerosol challenge with all three viruses. Single- and triple-virus-specific antibodies were isolated, and we identified 21 unique binding groups. Cryo-EM structures revealed that broad VLP binding inversely correlated with sequence and conformational variability. One triple-specific antibody, SKT05, bound proximal to the fusion peptide and neutralized all three Env-pseudotyped encephalitic alphaviruses by using different symmetry elements for recognition across VLPs. Neutralization in other assays (e.g., chimeric Sindbis virus) yielded variable results. SKT05 bound backbone atoms of sequence-diverse residues, enabling broad recognition despite sequence variability; accordingly, SKT05 protected mice against Venezuelan equine encephalitis virus, chikungunya virus, and Ross River virus challenges. Thus, a single vaccine-elicited antibody can protect in vivo against a broad range of alphaviruses.
History
DepositionJul 27, 2022-
Header (metadata) releaseJul 5, 2023-
Map releaseJul 5, 2023-
UpdateJul 5, 2023-
Current statusJul 5, 2023Processing site: RCSB / Status: Released

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Structure visualization

Supplemental images

emd_27723.png

Downloads & links

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Map

FileDownload / File: emd_27723.map.gz / Format: CCP4 / Size: 1.1 GB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Voxel sizeX=Y=Z: 1.66 Å
Density
Contour LevelBy AUTHOR: 0.3
Minimum - Maximum-1.0449824 - 1.6680474
Average (Standard dev.)0.0007619141 (±0.033410817)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions672672672
Spacing672672672
CellA=B=C: 1115.52 Å
α=β=γ: 90.0 °

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Supplemental data

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Half map: #2

Fileemd_27723_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_27723_half_map_2.map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Sample components

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Entire : Western Equine Encephalitis Virus trimer in complex with Antibody...

EntireName: Western Equine Encephalitis Virus trimer in complex with Antibody Fab SKW11
Components
  • Complex: Western Equine Encephalitis Virus trimer in complex with Antibody Fab SKW11
    • Protein or peptide: Antibody SKW11 heavy chain
    • Protein or peptide: Antibody SKW11 light chain
    • Protein or peptide: Spike glycoprotein E1
    • Protein or peptide: Spike glycoprotein E2
  • Ligand: 2-acetamido-2-deoxy-beta-D-glucopyranose

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Supramolecule #1: Western Equine Encephalitis Virus trimer in complex with Antibody...

SupramoleculeName: Western Equine Encephalitis Virus trimer in complex with Antibody Fab SKW11
type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#4
Source (natural)Organism: Western equine encephalitis virus

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Macromolecule #1: Antibody SKW11 heavy chain

MacromoleculeName: Antibody SKW11 heavy chain / type: protein_or_peptide / ID: 1 / Number of copies: 3 / Enantiomer: LEVO
Source (natural)Organism: Macaca fascicularis (crab-eating macaque)
Molecular weightTheoretical: 24.709627 KDa
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString: QVQLQESGPG LVKPSETLSL TCAVSGDSIS NNYWTWIRHF PGKGLEWIGR IYGDAGSTDY NPSLKSRVTI SMDLSKNQFS LDLTSVTVA DTALYYCASR TTVADNWFDV WGPGVLVTVS SASTKGPSVF PLAPSSRSTS ESTAALGCLV KDYFPEPVTV S WNSGSLTS ...String:
QVQLQESGPG LVKPSETLSL TCAVSGDSIS NNYWTWIRHF PGKGLEWIGR IYGDAGSTDY NPSLKSRVTI SMDLSKNQFS LDLTSVTVA DTALYYCASR TTVADNWFDV WGPGVLVTVS SASTKGPSVF PLAPSSRSTS ESTAALGCLV KDYFPEPVTV S WNSGSLTS GVHTFPAVLQ SSGLYSLSSV VTVPSSSLGT QTYVCNVNHK PSNTKVDKRV EIKTCGGLEV LFQ

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Macromolecule #2: Antibody SKW11 light chain

MacromoleculeName: Antibody SKW11 light chain / type: protein_or_peptide / ID: 2 / Number of copies: 3 / Enantiomer: LEVO
Source (natural)Organism: Macaca fascicularis (crab-eating macaque)
Molecular weightTheoretical: 23.317848 KDa
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString: DIQMTQSPSS LSASVGDTVT ITCRATQSIS SLLAWYQYKP GKAPKLLIYQ ASSLHIGVPS RFSGSGSGTD FTLTISSLQS EDFATYYCQ QHDSRPWTFG QGTKVDIKRT VAAPSVFIFP PSEDQVKSGT VSVVCLLNNF YPREASVKWK VDGALKTGNS Q ESVTEQDS ...String:
DIQMTQSPSS LSASVGDTVT ITCRATQSIS SLLAWYQYKP GKAPKLLIYQ ASSLHIGVPS RFSGSGSGTD FTLTISSLQS EDFATYYCQ QHDSRPWTFG QGTKVDIKRT VAAPSVFIFP PSEDQVKSGT VSVVCLLNNF YPREASVKWK VDGALKTGNS Q ESVTEQDS KDNTYSLSST LTLSSTEYQS HKVYACEVTH QGLSSPVTKS FNRGEC

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Macromolecule #3: Spike glycoprotein E1

MacromoleculeName: Spike glycoprotein E1 / type: protein_or_peptide / ID: 3 / Number of copies: 3 / Enantiomer: LEVO
Source (natural)Organism: Western equine encephalitis virus
Molecular weightTheoretical: 52.905348 KDa
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString: RPTNAETFGE TLNHLWFNNQ PFLWAQLCIP LAALVILFRC FSCCMPFLLV AGVCLGKVDA FEHATTVPNV PGIPYKALVE RAGYAPLNL EITVVSSELT PSTNKEYVTC RFHTVIPSPQ VKCCGSLECK ASSKADYTCR VFGGVYPFMW GGAQCFCDSE N TQLSEAYV ...String:
RPTNAETFGE TLNHLWFNNQ PFLWAQLCIP LAALVILFRC FSCCMPFLLV AGVCLGKVDA FEHATTVPNV PGIPYKALVE RAGYAPLNL EITVVSSELT PSTNKEYVTC RFHTVIPSPQ VKCCGSLECK ASSKADYTCR VFGGVYPFMW GGAQCFCDSE N TQLSEAYV EFAPDCTIDH AVALKVHTAA LKVGLRIVYG NTTAHLDTFV NGVTPGSSRD LKVIAGPISA AFSPFDHKVV IR KGLVYNY DFPEYGAMKP GAFGDIQASS LDATDIVART DIRLLKPSVK NIHVPYTQAV SGYEMWKNNS GRPLQETAPF GCK IEVEPL RASNCAYGHI PISIDIPDAA FVRSSESPTI LEVSCTVADC IYSADFGGSL TLQYKADREG HCPVHSHSTT AVLK EATTH VTAVGSITLH FSTSSPQANF IVSLCGKKTT CNAECKPPAD HIIGEPHKVD QEFQAAVSKT SWNWLLALFG GASSL IVVG LIVLV

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Macromolecule #4: Spike glycoprotein E2

MacromoleculeName: Spike glycoprotein E2 / type: protein_or_peptide / ID: 4 / Number of copies: 3 / Enantiomer: LEVO
Source (natural)Organism: Western equine encephalitis virus
Molecular weightTheoretical: 45.29373 KDa
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString: PYLGFCPYCR HSAPCFSPIK IENVWDESDD GSIRIQVSAQ FGYNQAGTAD VTKFRYMSFD HDHDIKEDSM DKIAISTSGP CRRLGHKGY FLLAQCPPGD SVTVSITSGA SENSCTVEKK IRRKFVGREE YLFPPVHGKL VKCHVYDHLK ETSAGYITMH R PGPHAYKS ...String:
PYLGFCPYCR HSAPCFSPIK IENVWDESDD GSIRIQVSAQ FGYNQAGTAD VTKFRYMSFD HDHDIKEDSM DKIAISTSGP CRRLGHKGY FLLAQCPPGD SVTVSITSGA SENSCTVEKK IRRKFVGREE YLFPPVHGKL VKCHVYDHLK ETSAGYITMH R PGPHAYKS YLEEASGEVY IKPPSGKNVT YECKCGDYST GIVSTRTKMN GCTKAKQCIA YKSDQTKWVF NSPDLIRHTD HS VQGKLHI PFRLTPTVCP VPLAHTPTVT KWFKGITLHL TATRPTLLTT RKLGLRADAT AEWITGTTSR NFSVGREGLE YVW GNHEPV RVWAQESAPG DPHGWPHEII IHYYHRHPVY TVIVLCGVAL AILVGTASSA ACIAKARRDC LTPYALAPNA TVPT ALAVL CCI

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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: 7 / Formula: NAG
Molecular weightTheoretical: 221.208 Da
Chemical component information

ChemComp-NAG:
2-acetamido-2-deoxy-beta-D-glucopyranose / N-Acetylglucosamine

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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: 7.4
VitrificationCryogen name: ETHANE

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Electron microscopy

MicroscopeTFS KRIOS
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: OTHER / Imaging mode: BRIGHT FIELDBright-field microscopy / Nominal defocus max: 2.0 µm / Nominal defocus min: 0.8 µm
Image recordingFilm or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Average electron dose: 46.5 e/Å2
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Startup modelType of model: NONE
Initial angle assignmentType: MAXIMUM LIKELIHOOD
Final angle assignmentType: MAXIMUM LIKELIHOOD
Final reconstructionResolution.type: BY AUTHOR / Resolution: 5.84 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 854520

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