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Open data
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Basic information
Entry | ![]() | |||||||||
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Title | Orf virus scaffolding protein Orfv075 | |||||||||
![]() | 3D reconstruction of Orfv075 (sharpened) | |||||||||
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![]() | Scaffold protein / Capsid protein / Poxvirus / VIRAL PROTEIN | |||||||||
Function / homology | Poxvirus rifampicin-resistance / Poxvirus rifampicin resistance protein / response to antibiotic / 62 kDa protein![]() | |||||||||
Biological species | ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 1.9 Å | |||||||||
![]() | Hyun J / Kim S / Ko S / Kim M / Jang Y | |||||||||
Funding support | ![]()
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![]() | ![]() Title: Cryo-EM structure of orf virus scaffolding protein orfv075. Authors: Seungmi Kim / Sumin Ko / Minjae Kim / Yeontae Jang / Jaekyung Hyun / ![]() Abstract: Capsid-like poxvirus scaffold proteins self-assemble into semi-regular lattice that govern the formation of spherical immature virus particles. The scaffolding is a critical step in virus ...Capsid-like poxvirus scaffold proteins self-assemble into semi-regular lattice that govern the formation of spherical immature virus particles. The scaffolding is a critical step in virus morphogenesis as exemplified by the drug rifampicin that impairs the recruitment of scaffold onto the viral membrane in vaccinia virus (VACV). Here we report cryo-electron microscopy structure of scaffolding protein Orfv075 of orf virus (ORFV) that causes smallpox-like diseases in sheep, goats and occasionally humans via zoonotic infection. We demonstrate that the regions that are involved in intertrimeric interactions for scaffold assembly are largely conserved in comparison to its VACV orthologue protein D13 whose intermediate assembly structures have been previously characterized. By contrast, less conserved regions are located away from these interfaces, indicating both viruses share similar assembly mechanisms. We also show that the phenylalanine-rich binding site of rifampicin in D13 is conserved in Orfv075, and molecular docking simulation confirms similar binding modes. Our study provides structural basis of scaffolding protein as a target for anti-poxvirus treatment across wide range of poxvirus genera. | |||||||||
History |
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Structure visualization
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 203.7 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 22.8 KB 22.8 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 13.6 KB | Display | ![]() |
Images | ![]() | 180.7 KB | ||
Masks | ![]() | 216 MB | ![]() | |
Filedesc metadata | ![]() | 7 KB | ||
Others | ![]() ![]() ![]() | 108.7 MB 200.7 MB 200.7 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 1017.6 KB | Display | ![]() |
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Full document | ![]() | 1017.2 KB | Display | |
Data in XML | ![]() | 20.6 KB | Display | |
Data in CIF | ![]() | 26.2 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 9ikcMC M: atomic model generated by this map C: citing same article ( |
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Similar structure data | Similarity search - Function & homology ![]() |
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Links
EMDB pages | ![]() ![]() |
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Map
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Annotation | 3D reconstruction of Orfv075 (sharpened) | ||||||||||||||||||||
Voxel size | X=Y=Z: 0.65 Å | ||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Mask #1
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Projections & Slices |
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Density Histograms |
-Additional map: 3D reconstruction of Orfv075 (unsharpened)
File | emd_60655_additional_1.map | ||||||||||||
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Annotation | 3D reconstruction of Orfv075 (unsharpened) | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: First halfmap of Orfv075 3D reconstruction
File | emd_60655_half_map_1.map | ||||||||||||
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Annotation | First halfmap of Orfv075 3D reconstruction | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Second halfmap of Orfv075 3D reconstruction
File | emd_60655_half_map_2.map | ||||||||||||
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Annotation | Second halfmap of Orfv075 3D reconstruction | ||||||||||||
Projections & Slices |
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Density Histograms |
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Sample components
-Entire : Orfv075 trimer
Entire | Name: Orfv075 trimer |
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Components |
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-Supramolecule #1: Orfv075 trimer
Supramolecule | Name: Orfv075 trimer / type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1 |
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Source (natural) | Organism: ![]() |
Molecular weight | Theoretical: 190 KDa |
-Macromolecule #1: 62 kDa protein
Macromolecule | Name: 62 kDa protein / type: protein_or_peptide / ID: 1 / Number of copies: 3 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() |
Molecular weight | Theoretical: 63.652172 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: MSYYHHHHHH DYDIPTTENL YFQGAMNNSV VSLVGGDDAV RRASVFATDH RAPTVYMPQY ITTQGVVDTT SDAVTVTFEI RDKYISAMN NFVLSVDLPE IKGVGKMCYV PYIAYKLIRH VAVNSAADTI WETSGEELFD SCLDNERVME LSGFSRELND L STGSSPND ...String: MSYYHHHHHH DYDIPTTENL YFQGAMNNSV VSLVGGDDAV RRASVFATDH RAPTVYMPQY ITTQGVVDTT SDAVTVTFEI RDKYISAMN NFVLSVDLPE IKGVGKMCYV PYIAYKLIRH VAVNSAADTI WETSGEELFD SCLDNERVME LSGFSRELND L STGSSPND VIKEAACVHA YIKTPFDADK TFSTLKLSDS KVTVTVTLNP VACVMVYDET FDAAKLAKEF PYSMELSFIG YM VKNLCPR PAFIEMPRRR VEQINHTTAV ITDVHACTSL SVYMKPVLSD ANNRFISYPG FQQSEGDFVM AFVERLLEDM VIV SNCYPE GFPETAEIVE VPPSGVVSIQ DTDVFVRIDD VPVGMRVFLH TNILVFATRK NSVVYNMSKK FSAITGAYSR ATSR IRFTT AIHSVNIGDA SVPVGVWTCQ RNVYNGDNRS PEARAKDLFV ADPFLKGVDF KNKIDVIARM DVRFGNEVLY SENSA VSRV FGEILGKTPG VRTLQFNFTP STFFSPTALN SNVSRGKDKL AVRVTTAHME AHNPLMYVPR QMVVVCNEVY RLSYDA GIV AEKVTAQ UniProtKB: 62 kDa protein |
-Macromolecule #2: water
Macromolecule | Name: water / type: ligand / ID: 2 / Number of copies: 150 / Formula: HOH |
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Molecular weight | Theoretical: 18.015 Da |
Chemical component information | ![]() ChemComp-HOH: |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Concentration | 0.13 mg/mL | ||||||||||||||||||
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Buffer | pH: 8 Component:
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Grid | Model: Quantifoil R1.2/1.3 / Material: COPPER / Mesh: 300 / Support film - Material: GRAPHENE OXIDE / Support film - topology: CONTINUOUS | ||||||||||||||||||
Vitrification | Cryogen name: ETHANE / Chamber humidity: 90 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK IV |
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Electron microscopy
Microscope | TFS KRIOS |
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Temperature | Min: 77.0 K / Max: 93.0 K |
Specialist optics | Energy filter - Name: GIF Bioquantum / Energy filter - Slit width: 20 eV |
Image recording | Film or detector model: GATAN K3 BIOCONTINUUM (6k x 4k) / Digitization - Dimensions - Width: 5760 pixel / Digitization - Dimensions - Height: 4092 pixel / Number grids imaged: 1 / Number real images: 10975 / Average exposure time: 2.6 sec. / Average electron dose: 50.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | C2 aperture diameter: 70.0 µm / Calibrated magnification: 76923 / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 1.8 µm / Nominal defocus min: 0.7000000000000001 µm / Nominal magnification: 130000 |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
-Atomic model buiding 1
Initial model | PDB ID: Chain - Chain ID: A / Chain - Residue range: 1-547 / Chain - Source name: PDB / Chain - Initial model type: experimental model |
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Details | Structure of orthologue protein D13 (PDB 7VFD) was fitted into the density map of orfv075 using UCSF Chimera. The fitted structure was modified in Coot based on sequence alignment between D13 and Orfv075. The resulting Orfv075 model was refined using PHENIX. |
Refinement | Space: REAL / Protocol: RIGID BODY FIT / Target criteria: Cross-correlation coefficient |
Output model | ![]() PDB-9ikc: |