+
Open data
-
Basic information
Entry | Database: EMDB / ID: EMD-23625 | |||||||||
---|---|---|---|---|---|---|---|---|---|---|
Title | HMPV F v3-B in complex with MPE33 Fab | |||||||||
![]() | ||||||||||
![]() |
| |||||||||
Function / homology | Precursor fusion glycoprotein F0, Paramyxoviridae / Fusion glycoprotein F0 / membrane => GO:0016020 / fusion of virus membrane with host plasma membrane / host cell plasma membrane / virion membrane / plasma membrane / Fusion glycoprotein F0![]() | |||||||||
Biological species | ![]() ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 4.79 Å | |||||||||
![]() | Gorman J / Kwong PD | |||||||||
Funding support | ![]()
| |||||||||
![]() | ![]() Title: Interprotomer disulfide-stabilized variants of the human metapneumovirus fusion glycoprotein induce high titer-neutralizing responses. Authors: Guillaume B E Stewart-Jones / Jason Gorman / Li Ou / Baoshan Zhang / M Gordon Joyce / Lijuan Yang / Cheng Cheng / Gwo-Yu Chuang / Kathryn E Foulds / Wing-Pui Kong / Adam S Olia / Mallika ...Authors: Guillaume B E Stewart-Jones / Jason Gorman / Li Ou / Baoshan Zhang / M Gordon Joyce / Lijuan Yang / Cheng Cheng / Gwo-Yu Chuang / Kathryn E Foulds / Wing-Pui Kong / Adam S Olia / Mallika Sastry / Chen-Hsiang Shen / John-Paul Todd / Yaroslav Tsybovsky / Raffaello Verardi / Yongping Yang / Peter L Collins / Davide Corti / Antonio Lanzavecchia / Diana G Scorpio / John R Mascola / Ursula J Buchholz / Peter D Kwong / ![]() ![]() Abstract: Human metapneumovirus (HMPV) is a major cause of respiratory disease worldwide, particularly among children and the elderly. Although there is no licensed HMPV vaccine, promising candidates have been ...Human metapneumovirus (HMPV) is a major cause of respiratory disease worldwide, particularly among children and the elderly. Although there is no licensed HMPV vaccine, promising candidates have been identified for related pneumoviruses based on the structure-based stabilization of the fusion (F) glycoprotein trimer, with prefusion-stabilized F glycoprotein trimers eliciting significantly higher neutralizing responses than their postfusion F counterparts. However, immunization with HMPV F trimers in either prefusion or postfusion conformations has been reported to elicit equivalent neutralization responses. Here we investigate the impact of stabilizing disulfides, especially interprotomer disulfides (IP-DSs) linking protomers of the F trimer, on the elicitation of HMPV-neutralizing responses. We designed F trimer disulfides, screened for their expression, and used electron microscopy (EM) to confirm their formation, including that of an unexpected postfusion variant. In mice, IP-DS-stabilized prefusion and postfusion HMPV F elicited significantly higher neutralizing responses than non-IP-DS-stabilized HMPV Fs. In macaques, the impact of IP-DS stabilization was more measured, although IP-DS-stabilized variants of either prefusion or postfusion HMPV F induced neutralizing responses many times the average titers observed in a healthy human cohort. Serological and absorption-based analyses of macaque responses revealed elicited HMPV-neutralizing responses to be absorbed differently by IP-DS-containing and by non-IP-DS-containing postfusion Fs, suggesting IP-DS stabilization to alter not only the immunogenicity of select epitopes but their antigenicity as well. We speculate the observed increase in immunogenicity by IP-DS trimers to be related to reduced interprotomer flexibility within the HMPV F trimer. | |||||||||
History |
|
-
Structure visualization
Movie |
![]() |
---|---|
Structure viewer | EM map: ![]() ![]() ![]() |
Supplemental images |
-
Downloads & links
-EMDB archive
Map data | ![]() | 230.2 MB | ![]() | |
---|---|---|---|---|
Header (meta data) | ![]() ![]() | 23.9 KB 23.9 KB | Display Display | ![]() |
Images | ![]() | 94 KB | ||
Masks | ![]() | 244.1 MB | ![]() | |
Others | ![]() ![]() ![]() ![]() | 120 MB 8.5 MB 226.4 MB 226.5 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
-
Links
EMDB pages | ![]() ![]() |
---|---|
Related items in Molecule of the Month |
-
Map
File | ![]() | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Voxel size | X=Y=Z: 1.0961 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
|
-Supplemental data
-Mask #1
File | ![]() | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Projections & Slices |
| ||||||||||||
Density Histograms |
-Additional map: unsharpened map
File | emd_23625_additional_1.map | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Annotation | unsharpened map | ||||||||||||
Projections & Slices |
| ||||||||||||
Density Histograms |
-Additional map: density modified map with ResolveCryoEM
File | emd_23625_additional_2.map | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Annotation | density modified map with ResolveCryoEM | ||||||||||||
Projections & Slices |
| ||||||||||||
Density Histograms |
-Half map: half map B
File | emd_23625_half_map_1.map | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Annotation | half map B | ||||||||||||
Projections & Slices |
| ||||||||||||
Density Histograms |
-Half map: half map A
File | emd_23625_half_map_2.map | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Annotation | half map A | ||||||||||||
Projections & Slices |
| ||||||||||||
Density Histograms |
-
Sample components
-Entire : HMPV F v3-B in complex with MPE33
Entire | Name: HMPV F v3-B in complex with MPE33 |
---|---|
Components |
|
-Supramolecule #1: HMPV F v3-B in complex with MPE33
Supramolecule | Name: HMPV F v3-B in complex with MPE33 / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
---|
-Supramolecule #2: HMPV F v3-B
Supramolecule | Name: HMPV F v3-B / type: complex / ID: 2 / Parent: 1 / Macromolecule list: #1 |
---|---|
Source (natural) | Organism: ![]() |
Recombinant expression | Organism: ![]() |
-Supramolecule #3: MPE33
Supramolecule | Name: MPE33 / type: complex / ID: 3 / Parent: 1 / Macromolecule list: #2-#3 |
---|---|
Source (natural) | Organism: ![]() |
Recombinant expression | Organism: ![]() |
-Macromolecule #1: HMPV F v3-B
Macromolecule | Name: HMPV F v3-B / type: protein_or_peptide / ID: 1 / Enantiomer: LEVO |
---|---|
Source (natural) | Organism: ![]() |
Recombinant expression | Organism: ![]() |
Sequence | String: TPQHGLKESY LEESCSTITE GYLSVLRTGW YTNVFTLEVG DVENLTCTDG PSLIKTELDL TKSALRELKT CSADQLAREE QIEGGGGGGF VLGAIALGVA TAAAVTAGIA IAKTIRLESE VNAIKGCLKT TNECVSTLGN GVRVLATAVR ELKEFVSKNL TSAINKNKCD ...String: TPQHGLKESY LEESCSTITE GYLSVLRTGW YTNVFTLEVG DVENLTCTDG PSLIKTELDL TKSALRELKT CSADQLAREE QIEGGGGGGF VLGAIALGVA TAAAVTAGIA IAKTIRLESE VNAIKGCLKT TNECVSTLGN GVRVLATAVR ELKEFVSKNL TSAINKNKCD IPDLKMAVSF SQFNRRFLNV VRQFSDNAGI TPAISLDLMT DAELARAVSY MPTSAGQIKL MLENRCMVRR KGFGILIGVY GSSVIYMVQL PIFGVIDTPC WIIKAAPSCS EKDGNYACLL REDQGWYCKN AGSTVYYPND KDCETRGDHV FCDTAAGINV AEQSRECNIN ISTTNYPCKV STGRHPISMV ALSPLGALVA CYKGVSCSIG SNRVGIIKQL PKGCSYITNQ DADTVTIDNT VYQLSKVEGE QHVIKGRPVS SSFDPIKFPE DQFNVALDQV FESIENSQAL VDQSNKILNS AESAIGGYIP EAPRDGQAYV RKDGEWVLLS TFLGGLVPR |
-Macromolecule #2: MPE33 Heavy chain
Macromolecule | Name: MPE33 Heavy chain / type: protein_or_peptide / ID: 2 / Enantiomer: LEVO |
---|---|
Source (natural) | Organism: ![]() |
Recombinant expression | Organism: ![]() |
Sequence | String: tgvhsqvqlq esgpglvkps qtlsltctvs gdsisngsyc wnwvrqpagk glewigriyp igntnynpsl ksrvtisldt sknqfslkla svtaadtavy ycarearife gyyyyyygld vwgqgttvtv ssastkgpsv fplapsskst sggtaalgcl vkdyfpepvt ...String: tgvhsqvqlq esgpglvkps qtlsltctvs gdsisngsyc wnwvrqpagk glewigriyp igntnynpsl ksrvtisldt sknqfslkla svtaadtavy ycarearife gyyyyyygld vwgqgttvtv ssastkgpsv fplapsskst sggtaalgcl vkdyfpepvt vswnsgalts gvhtfpavlq ssglyslssv vtvpssslgt qtyicnvnhk psntkvdkkv epkscdkglv pr |
-Macromolecule #3: MPE33 Light chain
Macromolecule | Name: MPE33 Light chain / type: protein_or_peptide / ID: 3 / Enantiomer: LEVO |
---|---|
Source (natural) | Organism: ![]() |
Recombinant expression | Organism: ![]() |
Sequence | String: EIVLTQSPAT LSLSPGERAT LSCRASQTIS SYLAWYQQKP GQAPRLLISD VSKRATGIPA RFSGSGSGTD FTLTISSLEP EDFAVYYCHQ RSNWDTFGQG TKLEIK |
-Experimental details
-Structure determination
Method | cryo EM |
---|---|
![]() | single particle reconstruction |
Aggregation state | particle |
-
Sample preparation
Concentration | 1.0 mg/mL |
---|---|
Buffer | pH: 7.4 / Component - Formula: PBS |
Grid | Model: C-flat-1.2/1.3 / Support film - Material: CARBON / Support film - topology: HOLEY |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 90 % / Chamber temperature: 293 K / Instrument: FEI VITROBOT MARK IV |
-
Electron microscopy
Microscope | FEI TITAN KRIOS |
---|---|
Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Number grids imaged: 1 / Average exposure time: 10.0 sec. / Average electron dose: 63.78 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | C2 aperture diameter: 70.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
+
Image processing
-Atomic model buiding 1
Initial model | PDB ID: |
---|---|
Refinement | Space: REAL |