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Yorodumi- EMDB-23640: Cryo-EM structure of the HCMV pentamer bound by antibodies 1-103,... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-23640 | |||||||||
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Title | Cryo-EM structure of the HCMV pentamer bound by antibodies 1-103, 1-32 and 2-25 | |||||||||
Map data | Focused refinement calculated in cryoSPARC and sharpened using DeepEMhancer. | |||||||||
Sample |
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Function / homology | Function and homology information host cell Golgi apparatus / entry receptor-mediated virion attachment to host cell / symbiont entry into host cell / fusion of virus membrane with host plasma membrane / viral envelope / host cell plasma membrane / virion membrane / membrane Similarity search - Function | |||||||||
Biological species | Human betaherpesvirus 5 / Homo sapiens (human) / Human cytomegalovirus | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.81 Å | |||||||||
Authors | Wrapp D / McLellan JS | |||||||||
Citation | Journal: Sci Adv / Year: 2022 Title: Structural basis for HCMV Pentamer recognition by neuropilin 2 and neutralizing antibodies. Authors: Daniel Wrapp / Xiaohua Ye / Zhiqiang Ku / Hang Su / Harrison G Jones / Nianshuang Wang / Akaash K Mishra / Daniel C Freed / Fengsheng Li / Aimin Tang / Leike Li / Dabbu Kumar Jaijyan / Hua ...Authors: Daniel Wrapp / Xiaohua Ye / Zhiqiang Ku / Hang Su / Harrison G Jones / Nianshuang Wang / Akaash K Mishra / Daniel C Freed / Fengsheng Li / Aimin Tang / Leike Li / Dabbu Kumar Jaijyan / Hua Zhu / Dai Wang / Tong-Ming Fu / Ningyan Zhang / Zhiqiang An / Jason S McLellan / Abstract: Human cytomegalovirus (HCMV) encodes multiple surface glycoprotein complexes to infect a variety of cell types. The HCMV Pentamer, composed of gH, gL, UL128, UL130, and UL131A, enhances entry into ...Human cytomegalovirus (HCMV) encodes multiple surface glycoprotein complexes to infect a variety of cell types. The HCMV Pentamer, composed of gH, gL, UL128, UL130, and UL131A, enhances entry into epithelial, endothelial, and myeloid cells by interacting with the cell surface receptor neuropilin 2 (NRP2). Despite the critical nature of this interaction, the molecular determinants that govern NRP2 recognition remain unclear. Here, we describe the cryo-EM structure of NRP2 bound to Pentamer. The high-affinity interaction between these proteins is calcium dependent and differs from the canonical carboxyl-terminal arginine (CendR) binding that NRP2 typically uses. We also determine the structures of four neutralizing human antibodies bound to the HCMV Pentamer to define susceptible epitopes. Two of these antibodies compete with NRP2 binding, but the two most potent antibodies recognize a previously unidentified epitope that does not overlap the NRP2-binding site. Collectively, these findings provide a structural basis for HCMV tropism and antibody-mediated neutralization. | |||||||||
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_23640.map.gz | 212.2 MB | EMDB map data format | |
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Header (meta data) | emd-23640-v30.xml emd-23640.xml | 33.4 KB 33.4 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_23640_fsc.xml | 15.5 KB | Display | FSC data file |
Images | emd_23640.png | 120.1 KB | ||
Others | emd_23640_additional_1.map.gz emd_23640_half_map_1.map.gz emd_23640_half_map_2.map.gz | 237.9 MB 285.2 MB 285.2 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-23640 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-23640 | HTTPS FTP |
-Validation report
Summary document | emd_23640_validation.pdf.gz | 715.9 KB | Display | EMDB validaton report |
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Full document | emd_23640_full_validation.pdf.gz | 715.5 KB | Display | |
Data in XML | emd_23640_validation.xml.gz | 23.2 KB | Display | |
Data in CIF | emd_23640_validation.cif.gz | 30.4 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-23640 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-23640 | HTTPS FTP |
-Related structure data
Related structure data | 7m30MC 7kbaC 7kbbC 7lyvC 7lywC 7m1cC 7m22C 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_23640.map.gz / Format: CCP4 / Size: 307.5 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | Focused refinement calculated in cryoSPARC and sharpened using DeepEMhancer. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.075 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Additional map: Full map that was later subjected to focused...
File | emd_23640_additional_1.map | ||||||||||||
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Annotation | Full map that was later subjected to focused refinement to calculate the primary map. Calculated in cryoSPARC and sharpened using DeepEMhancer. | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Half map A.
File | emd_23640_half_map_1.map | ||||||||||||
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Annotation | Half map A. | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Half map B.
File | emd_23640_half_map_2.map | ||||||||||||
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Annotation | Half map B. | ||||||||||||
Projections & Slices |
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Density Histograms |
-Sample components
+Entire : Quaternary complex of the HCMV pentamer bound by Fabs 1-103, 1-32...
+Supramolecule #1: Quaternary complex of the HCMV pentamer bound by Fabs 1-103, 1-32...
+Supramolecule #2: HCMV pentamer
+Supramolecule #3: 2-25 Fab
+Supramolecule #4: 1-32 Fab
+Supramolecule #5: 1-103 Fab
+Macromolecule #1: Envelope glycoprotein L
+Macromolecule #2: Envelope protein UL128
+Macromolecule #3: Envelope glycoprotein UL130
+Macromolecule #4: UL131A
+Macromolecule #5: 2-25 Fab Heavy Chain
+Macromolecule #6: 2-25 Fab Light Chain
+Macromolecule #7: 1-32 Fab Heavy Chain
+Macromolecule #8: 1-32 Fab Light Chain
+Macromolecule #9: 1-103 Fab Heavy Chain
+Macromolecule #10: 1-103 Fab Light Chain
+Macromolecule #11: 2-acetamido-2-deoxy-beta-D-glucopyranose
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Concentration | 0.2 mg/mL | |||||||||||||||
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Buffer | pH: 8 Component:
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Grid | Model: C-flat-1.2/1.3 / Material: COPPER / Support film - Material: CARBON / Support film - topology: HOLEY ARRAY / Pretreatment - Type: PLASMA CLEANING | |||||||||||||||
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277.15 K / Instrument: FEI VITROBOT MARK IV Details: Blotted for six seconds with a force of "1" before plunging. |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Average electron dose: 36.0 e/Å2 Details: Collected from a single grid at a mixture of 0 degrees tilt and -30 degrees tilt |
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 |