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Yorodumi- EMDB-12544: Hantaan virus-like particle glycoprotein spike in complex with Fa... -
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-Basic information
Entry | Database: EMDB / ID: EMD-12544 | |||||||||
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Title | Hantaan virus-like particle glycoprotein spike in complex with Fab fragment HTN-Gn1. | |||||||||
Map data | A reconstruction of a Hantaan virus-like particle glycoprotein spike bound to the Fab fragment of HTN-Gn1. | |||||||||
Sample |
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Function / homology | Function and homology information symbiont-mediated suppression of host TRAF-mediated signal transduction / host cell Golgi membrane / host cell mitochondrion / host cell surface / host cell endoplasmic reticulum membrane / virus-mediated perturbation of host defense response / virion membrane / signal transduction / membrane / metal ion binding Similarity search - Function | |||||||||
Biological species | Oryctolagus cuniculus (rabbit) / Hantaan orthohantavirus | |||||||||
Method | subtomogram averaging / cryo EM / Resolution: 19.0 Å | |||||||||
Authors | Stass R / Rissanen I / Bowden TA / Huiskonen JT | |||||||||
Funding support | United Kingdom, 1 items
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Citation | Journal: mBio / Year: 2021 Title: Structural Basis for a Neutralizing Antibody Response Elicited by a Recombinant Hantaan Virus Gn Immunogen. Authors: Ilona Rissanen / Stefanie A Krumm / Robert Stass / Annalis Whitaker / James E Voss / Emily A Bruce / Sylvia Rothenberger / Stefan Kunz / Dennis R Burton / Juha T Huiskonen / Jason W Botten / ...Authors: Ilona Rissanen / Stefanie A Krumm / Robert Stass / Annalis Whitaker / James E Voss / Emily A Bruce / Sylvia Rothenberger / Stefan Kunz / Dennis R Burton / Juha T Huiskonen / Jason W Botten / Thomas A Bowden / Katie J Doores / Abstract: Hantaviruses are a group of emerging pathogens capable of causing severe disease upon zoonotic transmission to humans. The mature hantavirus surface presents higher-order tetrameric assemblies of two ...Hantaviruses are a group of emerging pathogens capable of causing severe disease upon zoonotic transmission to humans. The mature hantavirus surface presents higher-order tetrameric assemblies of two glycoproteins, Gn and Gc, which are responsible for negotiating host cell entry and constitute key therapeutic targets. Here, we demonstrate that recombinantly derived Gn from Hantaan virus (HTNV) elicits a neutralizing antibody response (serum dilution that inhibits 50% infection [ID], 1:200 to 1:850) in an animal model. Using antigen-specific B cell sorting, we isolated monoclonal antibodies (mAbs) exhibiting neutralizing and non-neutralizing activity, termed mAb HTN-Gn1 and mAb nnHTN-Gn2, respectively. Crystallographic analysis reveals that these mAbs target spatially distinct epitopes at disparate sites of the N-terminal region of the HTNV Gn ectodomain. Epitope mapping onto a model of the higher order (Gn-Gc) spike supports the immune accessibility of the mAb HTN-Gn1 epitope, a hypothesis confirmed by electron cryo-tomography of the antibody with virus-like particles. These data define natively exposed regions of the hantaviral Gn that can be targeted in immunogen design. The spillover of pathogenic hantaviruses from rodent reservoirs into the human population poses a continued threat to human health. Here, we show that a recombinant form of the Hantaan virus (HTNV) surface-displayed glycoprotein, Gn, elicits a neutralizing antibody response in rabbits. We isolated a neutralizing (HTN-Gn1) and a non-neutralizing (nnHTN-Gn2) monoclonal antibody and provide the first molecular-level insights into how the Gn glycoprotein may be targeted by the antibody-mediated immune response. These findings may guide rational vaccine design approaches focused on targeting the hantavirus glycoprotein envelope. | |||||||||
History |
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-Structure visualization
Movie |
Movie viewer |
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Structure viewer | EM map: SurfViewMolmilJmol/JSmol |
Supplemental images |
-Downloads & links
-EMDB archive
Map data | emd_12544.map.gz | 847 KB | EMDB map data format | |
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Header (meta data) | emd-12544-v30.xml emd-12544.xml | 17.1 KB 17.1 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_12544_fsc.xml | 2.5 KB | Display | FSC data file |
Images | emd_12544.png | 98.7 KB | ||
Masks | emd_12544_msk_1.map | 932 KB | Mask map | |
Others | emd_12544_half_map_1.map.gz emd_12544_half_map_2.map.gz | 840 KB 445.6 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-12544 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-12544 | HTTPS FTP |
-Related structure data
Related structure data | 7nrhMC 7nksC 7o9sC M: atomic model generated by this map C: citing same article (ref.) |
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Similar structure data |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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-Map
File | Download / File: emd_12544.map.gz / Format: CCP4 / Size: 931.6 KB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | A reconstruction of a Hantaan virus-like particle glycoprotein spike bound to the Fab fragment of HTN-Gn1. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 7.04 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Mask #1
File | emd_12544_msk_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: Half map (odd) used to estimate resolution by FSC.
File | emd_12544_half_map_1.map | ||||||||||||
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Annotation | Half map (odd) used to estimate resolution by FSC. | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Half map (even) used to estimate resolution by FSC.
File | emd_12544_half_map_2.map | ||||||||||||
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Annotation | Half map (even) used to estimate resolution by FSC. | ||||||||||||
Projections & Slices |
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Density Histograms |
-Sample components
-Entire : Hantaan orthohantavirus
Entire | Name: Hantaan orthohantavirus |
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Components |
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-Supramolecule #1: Hantaan orthohantavirus
Supramolecule | Name: Hantaan orthohantavirus / type: virus / ID: 1 / Parent: 0 / Macromolecule list: all / NCBI-ID: 1980471 / Sci species name: Hantaan orthohantavirus / Virus type: VIRUS-LIKE PARTICLE / Virus isolate: SPECIES / Virus enveloped: Yes / Virus empty: Yes |
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Host system | Organism: Homo sapiens (human) |
-Macromolecule #1: Fab fragment HTN-Gn1 Heavy chain
Macromolecule | Name: Fab fragment HTN-Gn1 Heavy chain / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: Oryctolagus cuniculus (rabbit) |
Molecular weight | Theoretical: 24.284264 KDa |
Recombinant expression | Organism: Homo sapiens (human) |
Sequence | String: TGQSLVESGG DLVKPEGSLT LTCTASGFSF SSTHWICWVR QAPGKGLEWI ACIYVGNTYD SYYANWAKGR FTISKTSSTT VTLQMTTLT AADTATYFCA RSGSVFGVVS LWGPGTLVTV SSGQPKAPSV FPLAPCCGDT PSSTVTLGCL VKGYLPEPVT V TWNSGTLT ...String: TGQSLVESGG DLVKPEGSLT LTCTASGFSF SSTHWICWVR QAPGKGLEWI ACIYVGNTYD SYYANWAKGR FTISKTSSTT VTLQMTTLT AADTATYFCA RSGSVFGVVS LWGPGTLVTV SSGQPKAPSV FPLAPCCGDT PSSTVTLGCL VKGYLPEPVT V TWNSGTLT NGVRTFPSVR QSSGLYSLSS VVSVTSSSQP VTCNVAHPAT NTKVDKTVAP STCSGTKHHH HHH |
-Macromolecule #2: Fab fragment HTN-Gn1 Light chain
Macromolecule | Name: Fab fragment HTN-Gn1 Light chain / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: Oryctolagus cuniculus (rabbit) |
Molecular weight | Theoretical: 22.943162 KDa |
Recombinant expression | Organism: Homo sapiens (human) |
Sequence | String: TGQVLTQTPA SVSEPVEGTV TIKCQASQSI NNWLSWYQQR PGQPPKLLIY DASTVASGVS SRFKGSGSGT EFTLTISDLE CADAATYAC QSYGYGISIT DNSAFGGGTE VVVRGDPVAP SVLIFPPAAD QVATGTVTIV CVANKYFPDV TVTWEVDGTT Q TTGIENSK ...String: TGQVLTQTPA SVSEPVEGTV TIKCQASQSI NNWLSWYQQR PGQPPKLLIY DASTVASGVS SRFKGSGSGT EFTLTISDLE CADAATYAC QSYGYGISIT DNSAFGGGTE VVVRGDPVAP SVLIFPPAAD QVATGTVTIV CVANKYFPDV TVTWEVDGTT Q TTGIENSK TPQNSADCTY NLSSTLTLTS TQYNSHKEYT CKVTQGTTSV VQSFNRGDC |
-Macromolecule #3: Envelope polyprotein
Macromolecule | Name: Envelope polyprotein / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: Hantaan orthohantavirus |
Molecular weight | Theoretical: 40.350871 KDa |
Recombinant expression | Organism: Homo sapiens (human) |
Sequence | String: TGSLRNVYDM KIECPHTVSF GENSVIGYVE LPPMPLADTA QMVPESSCSM DNHQSINTIT KYTQVIWRGK ADPGQSSQNS FETVSTEVD LKGTCVLKHK MVEESYRSRK SITCYDLSCN STFCKPTLYM IVPIHACNMM KSCLIALGPY RVQVVYERTY C MTGVLIEG ...String: TGSLRNVYDM KIECPHTVSF GENSVIGYVE LPPMPLADTA QMVPESSCSM DNHQSINTIT KYTQVIWRGK ADPGQSSQNS FETVSTEVD LKGTCVLKHK MVEESYRSRK SITCYDLSCN STFCKPTLYM IVPIHACNMM KSCLIALGPY RVQVVYERTY C MTGVLIEG KCFVPDQSVV SIIKHGIFDI ASVHVVCFFV AVKGNTYKLF EQVKKSFEST CNDTENKVQG YYICIVGGNS AP IYVPTLD DFRSMEAFTG IFKSPHGEDH DLAGEEIASY SIVGPANAKV PHSASSDTLS LIAYSGIPSY SSLSILTSST DAK HVFSPG LFPKLNHTNC DKSAIPLTWT GMIDLPGYYE GTKHHHHHH |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | subtomogram averaging |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 7.5 / Details: PBS |
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Vitrification | Cryogen name: ETHANE-PROPANE |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Average electron dose: 4.77 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |