Journal: J Virol / Year: 2012 Title: Structural basis for broad detection of genogroup II noroviruses by a monoclonal antibody that binds to a site occluded in the viral particle. Authors: Grant S Hansman / David W Taylor / Jason S McLellan / Thomas J Smith / Ivelin Georgiev / Jeremy R H Tame / Sam-Yong Park / Makoto Yamazaki / Fumio Gondaira / Motohiro Miki / Kazuhiko ...Authors: Grant S Hansman / David W Taylor / Jason S McLellan / Thomas J Smith / Ivelin Georgiev / Jeremy R H Tame / Sam-Yong Park / Makoto Yamazaki / Fumio Gondaira / Motohiro Miki / Kazuhiko Katayama / Kazuyoshi Murata / Peter D Kwong / Abstract: Human noroviruses are genetically and antigenically highly divergent. Monoclonal antibodies raised in mice against one kind of norovirus virus-like particle (VLP), however, were found to have broad ...Human noroviruses are genetically and antigenically highly divergent. Monoclonal antibodies raised in mice against one kind of norovirus virus-like particle (VLP), however, were found to have broad recognition. In this study, we present the crystal structure of the antigen-binding fragment (Fab) for one of these broadly reactive monoclonal antibodies, 5B18, in complex with the capsid-protruding domain from a genogroup II genotype 10 (GII.10) norovirus at 3.3-Å resolution and, also, the cryo-electron microscopy structure of the GII.10 VLP at ∼10-Å resolution. The GII.10 VLP structure was more similar in overall architecture to the GV.1 murine norovirus virion than to the prototype GI.1 human norovirus VLP, with the GII.10 protruding domain raised ∼15 Å off the shell domain and rotated ∼40° relative to the GI.1 protruding domain. In the crystal structure, the 5B18 Fab bound to a highly conserved region of the protruding domain. Based on the VLP structure, this region is involved in interactions with other regions of the capsid and is buried in the virus particle. Despite the occluded nature of the recognized epitope in the VLP structure, enzyme-linked immunosorbent assay (ELISA) binding suggested that the 5B18 antibody was able to capture intact VLPs. Together, the results provide evidence that the norovirus particle is capable of extreme conformational flexibility, which may allow for antibody recognition of conserved surfaces that would otherwise be buried on intact particles.
History
Deposition
Dec 20, 2011
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Header (metadata) release
Jan 9, 2012
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Map release
Jan 30, 2012
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Update
Feb 8, 2012
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Current status
Feb 8, 2012
Processing site: RCSB / Status: Released
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Structure visualization
Movie
Surface view with section colored by density value
Organism: Homo sapiens (human) / synonym: VERTEBRATES
Molecular weight
Experimental: 10 MDa / Theoretical: 10 MDa
Virus shell
Shell ID: 1 / T number (triangulation number): 3
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Experimental details
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Structure determination
Method
cryo EM
Processing
single particle reconstruction
Aggregation state
particle
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Sample preparation
Concentration
1.0 mg/mL
Buffer
pH: 7.3 / Details: PBS
Grid
Details: Quantifoil R1.2/1.3 Mo 200 mesh
Vitrification
Cryogen name: ETHANE / Chamber humidity: 100 % / Instrument: FEI VITROBOT MARK IV Details: Vitrification instrument: FEI Vitrobot Mark IV. Quantifoil Mo 200 mesh holey carbon grid with a thin layer of carbon over the holes Method: automatic blotting for 8 seconds
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Electron microscopy
Microscope
JEOL 2200FS
Date
Jul 22, 2011
Image recording
Category: CCD / Film or detector model: GENERIC TVIPS (4k x 4k) / Average electron dose: 20 e/Å2
Electron beam
Acceleration voltage: 200 kV / Electron source: FIELD EMISSION GUN
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