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- EMDB-43894: HIV-1 B41 SOSIP.v4.1 Env in Complex with N241 Epitope and Anti-Im... -

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Basic information

Entry
Database: EMDB / ID: EMD-43894
TitleHIV-1 B41 SOSIP.v4.1 Env in Complex with N241 Epitope and Anti-Immune Complex Antibodies
Map data
Sample
  • Complex: HIV-1 B41 SOSIP Env in Complex with Rabbit Polyclonal Antibodies - N241 Epitope and Anti-Immune Complex pAb
    • Protein or peptide: HIV-1 B41 SOSIP.v4.1 Env - gp160
KeywordsHIV / Polyclonal / Antibodies / CryoEMPEM / Rabbit / Anti-Immune Complex / V1/V3 / VIRAL PROTEIN
Biological speciesOryctolagus cuniculus (rabbit) / Human immunodeficiency virus 1
Methodsingle particle reconstruction / cryo EM / Resolution: 5.8 Å
AuthorsBrown S / Antansijevic A / Ward AB
Funding support United States, 2 items
OrganizationGrant numberCountry
National Institutes of Health/National Institute Of Allergy and Infectious Diseases (NIH/NIAID)R01 AI136621 United States
Bill & Melinda Gates FoundationINV-002916 United States
CitationJournal: Sci Immunol / Year: 2025
Title: Anti-immune complex antibodies are elicited during repeated immunization with HIV Env immunogens.
Authors: Sharidan Brown / Aleksandar Antanasijevic / Leigh M Sewall / Daniel Montiel Garcia / James Ferguson / Philip J M Brouwer / Rogier W Sanders / Andrew B Ward /
Abstract: Vaccination strategies against HIV-1 aim to elicit broadly neutralizing antibodies (bnAbs) using prime-boost regimens with HIV envelope (Env) immunogens. Epitope mapping has shown that early antibody ...Vaccination strategies against HIV-1 aim to elicit broadly neutralizing antibodies (bnAbs) using prime-boost regimens with HIV envelope (Env) immunogens. Epitope mapping has shown that early antibody responses are directed to easily accessible nonneutralizing epitopes on Env instead of bnAb epitopes. Autologously neutralizing antibody responses appear upon boosting, once immunodominant epitopes are saturated. Here, we use electron microscopy-based polyclonal epitope mapping (EMPEM) to elucidate how repeated immunization with HIV Env SOSIP immunogens results in the generation of Ab2α anti-idiotypic antibodies in rabbits and rhesus macaques. We present the structures of six anti-immune complex antibodies and find that they target idiotopes composed of framework regions of antibodies bound to Env. Examination of cryo-electron microscopy density enabled prediction of sequences for an anti-immune complex antibody, the paratope of which is enriched with aromatic amino acids. This work sheds light on current vaccine development efforts for HIV, as well as for other pathogens in which repeated exposure to antigen is required.
History
DepositionFeb 29, 2024-
Header (metadata) releaseJan 29, 2025-
Map releaseJan 29, 2025-
UpdateJan 29, 2025-
Current statusJan 29, 2025Processing site: RCSB / Status: Released

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Structure visualization

Supplemental images

Downloads & links

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Map

FileDownload / File: emd_43894.map.gz / Format: CCP4 / Size: 178 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Projections & slices

Image control

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AxesZ (Sec.)Y (Row.)X (Col.)
1.15 Å/pix.
x 360 pix.
= 414. Å
1.15 Å/pix.
x 360 pix.
= 414. Å
1.15 Å/pix.
x 360 pix.
= 414. Å

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Images are generated by Spider.

Voxel sizeX=Y=Z: 1.15 Å
Density
Contour LevelBy AUTHOR: 0.009
Minimum - Maximum-0.008255402 - 0.022076149
Average (Standard dev.)0.00006632823 (±0.0008242848)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions360360360
Spacing360360360
CellA=B=C: 414.0 Å
α=β=γ: 90.0 °

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Supplemental data

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Mask #1

Fileemd_43894_msk_1.map
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Half map: #1

Fileemd_43894_half_map_1.map
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Half map: #2

Fileemd_43894_half_map_2.map
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Sample components

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Entire : HIV-1 B41 SOSIP Env in Complex with Rabbit Polyclonal Antibodies ...

EntireName: HIV-1 B41 SOSIP Env in Complex with Rabbit Polyclonal Antibodies - N241 Epitope and Anti-Immune Complex pAb
Components
  • Complex: HIV-1 B41 SOSIP Env in Complex with Rabbit Polyclonal Antibodies - N241 Epitope and Anti-Immune Complex pAb
    • Protein or peptide: HIV-1 B41 SOSIP.v4.1 Env - gp160

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Supramolecule #1: HIV-1 B41 SOSIP Env in Complex with Rabbit Polyclonal Antibodies ...

SupramoleculeName: HIV-1 B41 SOSIP Env in Complex with Rabbit Polyclonal Antibodies - N241 Epitope and Anti-Immune Complex pAb
type: complex / ID: 1 / Parent: 0 / Macromolecule list: all
Source (natural)Organism: Oryctolagus cuniculus (rabbit)

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Macromolecule #1: HIV-1 B41 SOSIP.v4.1 Env - gp160

MacromoleculeName: HIV-1 B41 SOSIP.v4.1 Env - gp160 / type: protein_or_peptide / ID: 1 / Enantiomer: LEVO
Source (natural)Organism: Human immunodeficiency virus 1
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString: MDAMKRGLCC VLLLCGAVFV SPSQEIHARF RRGARAAKKW VTVYYGVPVW KEATTTLFCA SDAKAYDTEV HNVWATHACV PTDPNPQEIV LGNVTENFNM WKNNMVEQMH EDIISLWDQS LKPCVKLTPL CVTLNCNNVN TNNTNNSTNA TISDWEKMET GEMKNCSFNV ...String:
MDAMKRGLCC VLLLCGAVFV SPSQEIHARF RRGARAAKKW VTVYYGVPVW KEATTTLFCA SDAKAYDTEV HNVWATHACV PTDPNPQEIV LGNVTENFNM WKNNMVEQMH EDIISLWDQS LKPCVKLTPL CVTLNCNNVN TNNTNNSTNA TISDWEKMET GEMKNCSFNV TTSIRDKIKK EYALFYKLDV VPLENKNNIN NTNITNYRLI NCNTSVITQA CPKVSFEPIP IHYCAPAGFA ILKCNSKTFN GSGPCTNVST VQCTHGIRPV VSTQLLLNGS LAEEEIVIRS ENITDNAKTI IVQLNEAVEI NCTRPNNNTR KSIHIGPGRA FYATGDIIGN IRQAHCNISK ARWNETLGQI VAKLEEQFPN KTIIFNHSSG GDPEIVTHSF NCGGEFFYCN TTPLFNSTWN NTRTDDYPTG GEQNITLQCR IKQIINMWQG VGKAMYAPPI RGQIRCSSNI TGLLLTRDGG RDQNGTETFR PGGGNMRDNW RSELYKYKVV KIEPLGIAPT ACKRRVQRRR RRRAVGLGAF ILGFLGAAGS TMGAASMALT VQARLLLSGI VQQQNNLLRA PEAQQHMLQL TVWGIKQLQA RVLAVERYLR DQQLLGIWGC SGKIICCTNV PWNDSWSNKT INEIWDNMTW MQWEKEIDNY TQHIYTLLEV SQIQQEKNEQ ELLELD

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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

Concentration5.8 mg/mL
BufferpH: 7.4
GridModel: Quantifoil R1.2/1.3 / Material: COPPER / Mesh: 300 / Support film - Material: CARBON / Support film - topology: HOLEY
VitrificationCryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 283 K / Instrument: FEI VITROBOT MARK IV

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Electron microscopy

MicroscopeFEI TALOS ARCTICA
Image recordingFilm or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Average electron dose: 50.2 e/Å2
Electron beamAcceleration voltage: 200 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 1.8 µm / Nominal defocus min: 1.0 µm
Experimental equipment
Model: Talos Arctica / Image courtesy: FEI Company

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Image processing

Startup modelType of model: INSILICO MODEL / In silico model: Ab initio model generated using CryoSPARC.
Final reconstructionResolution.type: BY AUTHOR / Resolution: 5.8 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: RELION (ver. 3.0) / Number images used: 33100
Initial angle assignmentType: MAXIMUM LIKELIHOOD
Final angle assignmentType: MAXIMUM LIKELIHOOD
FSC plot (resolution estimation)

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