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- EMDB-22988: Adeno-associated virus serotype 5 in complex with the cellular re... -
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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-22988 | |||||||||
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Title | Adeno-associated virus serotype 5 in complex with the cellular receptor AAVR at 2.5 Angstroms resolution, AAV5 AAVR | |||||||||
![]() | Adeno-associated virus serotype 5 bound to its cellular receptor AAVR, cryo-em sharpened map at 2.5 Angstroms resolution. | |||||||||
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![]() | AAV5 / AAV / AAV-5 / AAVR / Adeno Associated Virus / PKD domain / VIRUS LIKE PARTICLE / parvovirus / virus / gene therapy / Receptor / Adeno-associated virus receptor / PKD1 / PKD | |||||||||
Function / homology | ![]() T=1 icosahedral viral capsid / neuron migration / cytoplasmic vesicle / Golgi membrane / nucleolus / structural molecule activity / Golgi apparatus / membrane / plasma membrane Similarity search - Function | |||||||||
Biological species | ![]() ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 2.5 Å | |||||||||
![]() | Silveria M / Chapman MS | |||||||||
Funding support | ![]()
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![]() | ![]() Title: The Structure of an AAV5-AAVR Complex at 2.5 Å Resolution: Implications for Cellular Entry and Immune Neutralization of AAV Gene Therapy Vectors. Authors: Mark A Silveria / Edward E Large / Grant M Zane / Tommi A White / Michael S Chapman / ![]() Abstract: Adeno-Associated Virus is the leading vector for gene therapy. Although it is the vector for all in vivo gene therapies approved for clinical use by the US Food and Drug Administration, its biology ...Adeno-Associated Virus is the leading vector for gene therapy. Although it is the vector for all in vivo gene therapies approved for clinical use by the US Food and Drug Administration, its biology is still not yet fully understood. It has been shown that different serotypes of AAV bind to their cellular receptor, AAVR, in different ways. Previously we have reported a 2.4Å structure of AAV2 bound to AAVR that shows ordered structure for only one of the two AAVR domains with which AAV2 interacts. In this study we present a 2.5Å resolution structure of AAV5 bound to AAVR. AAV5 binds to the first polycystic kidney disease (PKD) domain of AAVR that was not ordered in the AAV2 structure. Interactions of AAV5 with AAVR are analyzed in detail, and the implications for AAV2 binding are explored through molecular modeling. Moreover, we find that binding sites for the antibodies ADK5a, ADK5b, and 3C5 on AAV5 overlap with the binding site of AAVR. These insights provide a structural foundation for development of gene therapy agents to better evade immune neutralization without disrupting cellular entry. | |||||||||
History |
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Structure visualization
Movie |
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Structure viewer | EM map: ![]() ![]() ![]() |
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 106.9 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 24.2 KB 24.2 KB | Display Display | ![]() |
Images | ![]() | 260.9 KB | ||
Masks | ![]() | 512 MB | ![]() | |
Filedesc metadata | ![]() | 7.3 KB | ||
Others | ![]() ![]() ![]() | 423.1 MB 432.7 MB 432.8 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 1.1 MB | Display | ![]() |
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Full document | ![]() | 1.1 MB | Display | |
Data in XML | ![]() | 19.5 KB | Display | |
Data in CIF | ![]() | 23 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 7kpnMC ![]() 7kp3C C: citing same article ( M: atomic model generated by this map |
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Similar structure data |
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Links
EMDB pages | ![]() ![]() |
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Related items in Molecule of the Month |
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Map
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Annotation | Adeno-associated virus serotype 5 bound to its cellular receptor AAVR, cryo-em sharpened map at 2.5 Angstroms resolution. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 0.66515 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Mask #1
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Projections & Slices |
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Density Histograms |
-Additional map: Adeno-associated virus serotype 5 bound to its cellular...
File | emd_22988_additional_1.map | ||||||||||||
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Annotation | Adeno-associated virus serotype 5 bound to its cellular receptor AAVR, cryo-em unsharpened map at 2.5 Angstroms resolution. | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Adeno-associated virus serotype 5 bound to its cellular...
File | emd_22988_half_map_1.map | ||||||||||||
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Annotation | Adeno-associated virus serotype 5 bound to its cellular receptor AAVR, cryo-em half map 2 of 2 for a map at 2.5 Angstroms resolution. | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Adeno-associated virus serotype 5 bound to its cellular...
File | emd_22988_half_map_2.map | ||||||||||||
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Annotation | Adeno-associated virus serotype 5 bound to its cellular receptor AAVR, cryo-em half map 1 of 2 for a map at 2.5 Angstroms resolution. | ||||||||||||
Projections & Slices |
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Density Histograms |
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Sample components
-Entire : Adeno-associated virus - 5
Entire | Name: ![]() |
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Components |
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-Supramolecule #1: Adeno-associated virus - 5
Supramolecule | Name: Adeno-associated virus - 5 / type: virus / ID: 1 / Parent: 0 / Macromolecule list: all Details: Expressed using SF9 cells with a pfastbac LIC vector. Purified with cesium chloride ultracentrifugation. NCBI-ID: 82300 / Sci species name: Adeno-associated virus - 5 / Virus type: VIRUS-LIKE PARTICLE / Virus isolate: SEROTYPE / Virus enveloped: No / Virus empty: Yes |
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Host (natural) | Organism: ![]() |
Molecular weight | Theoretical: 3.746 MDa |
Virus shell | Shell ID: 1 / Name: VP3 / Diameter: 250.0 Å / T number (triangulation number): 1 |
-Macromolecule #1: Dyslexia-associated protein KIAA0319-like protein
Macromolecule | Name: Dyslexia-associated protein KIAA0319-like protein / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() |
Molecular weight | Theoretical: 22.182744 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: MASHHHHHHV SAGESVQITL PKNEVQLNAY VLQEPPKGET YTYDWQLITH PRDYSGEMEG KHSQILKLSK LTPGLYEFKV IVEGQNAHG EGYVNVTVKP EPRKNRPPIA IVSPQFQEIS LPTTSTVIDG SQSTDDDKIV QYHWEELKGP LREEKISEDT A ILKLSKLV ...String: MASHHHHHHV SAGESVQITL PKNEVQLNAY VLQEPPKGET YTYDWQLITH PRDYSGEMEG KHSQILKLSK LTPGLYEFKV IVEGQNAHG EGYVNVTVKP EPRKNRPPIA IVSPQFQEIS LPTTSTVIDG SQSTDDDKIV QYHWEELKGP LREEKISEDT A ILKLSKLV PGNYTFSLTV VDSDGATNST TANLTVNKAV D UniProtKB: Dyslexia-associated protein KIAA0319-like protein |
-Macromolecule #2: Capsid protein
Macromolecule | Name: Capsid protein / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() |
Molecular weight | Theoretical: 80.366211 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: SFVDHPPDWL EEVGEGLREF LGLEAGPPKP KPNQQHQDQA RGLVLPGYNY LGPGNGLDRG EPVNRADEVA REHDISYNEQ LEAGDNPYL KYNHADAEFQ EKLADDTSFG GNLGKAVFQA KKRVLEPFGL VEEGAKTAPT GKRIDDHFPK RKKARTEEDS K PSTSSDAE ...String: SFVDHPPDWL EEVGEGLREF LGLEAGPPKP KPNQQHQDQA RGLVLPGYNY LGPGNGLDRG EPVNRADEVA REHDISYNEQ LEAGDNPYL KYNHADAEFQ EKLADDTSFG GNLGKAVFQA KKRVLEPFGL VEEGAKTAPT GKRIDDHFPK RKKARTEEDS K PSTSSDAE AGPSGSQQLQ IPAQPASSLG ADTMSAGGGG PLGDNNQGAD GVGNASGDWH CDSTWMGDRV VTKSTRTWVL PS YNNHQYR EIKSGSVDGS NANAYFGYST PWGYFDFNRF HSHWSPRDWQ RLINNYWGFR PRSLRVKIFN IQVKEVTVQD STT TIANNL TSTVQVFTDD DYQLPYVVGN GTEGCLPAFP PQVFTLPQYG YATLNRDNTE NPTERSSFFC LEYFPSKMLR TGNN FEFTY NFEEVPFHSS FAPSQNLFKL ANPLVDQYLY RFVSTNNTGG VQFNKNLAGR YANTYKNWFP GPMGRTQGWN LGSGV NRAS VSAFATTNRM ELEGASYQVP PQPNGMTNNL QGSNTYALEN TMIFNSQPAN PGTTATYLEG NMLITSESET QPVNRV AYN VGGQMATNNQ SSTTAPATGT YNLQEIVPGS VWMERDVYLQ GPIWAKIPET GAHFHPSPAM GGFGLKHPPP MMLIKNT PV PGNITSFSDV PVSSFITQYS TGQVTVEMEW ELKKENSKRW NPEIQYTNNY NDPQFVDFAP DSTGEYRTTR PIGTRYLT R PL UniProtKB: Capsid protein |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Concentration | 0.75 mg/mL | ||||||||||||
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Buffer | pH: 7.4 Component:
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Grid | Model: PELCO Ultrathin Carbon with Lacey Carbon / Material: COPPER / Pretreatment - Type: GLOW DISCHARGE | ||||||||||||
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 293 K / Instrument: FEI VITROBOT MARK IV Details: Two 2uL aliquots applied to grid (manual blotting between), prior to automated 3 second blot before plunging.. | ||||||||||||
Details | Monodisperse |
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Electron microscopy
Microscope | FEI TITAN KRIOS |
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Temperature | Min: 93.0 K / Max: 93.0 K |
Image recording | Film or detector model: GATAN K3 (6k x 4k) / Detector mode: INTEGRATING / Digitization - Dimensions - Width: 11520 pixel / Digitization - Dimensions - Height: 8184 pixel / Number grids imaged: 1 / Number real images: 734 / Average electron dose: 32.9 e/Å2 / Details: Pixel size was 0.664 angstrom. |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: -2.7 µm / Nominal defocus min: -0.7000000000000001 µm / Nominal magnification: 64000 |
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 - Source name: PDB / Chain - Initial model type: experimental model |
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Details | Stand-alone RSRef was used for refinement of magnification, resolution, envelope correction and atomic B-factors. This was alternated with RSRef-embedded CNS was used for molecular dynamics optimization (1st round) and stereochemically-restrained all-atom least-squares optimization. |
Refinement | Space: REAL / Protocol: FLEXIBLE FIT / Target criteria: Least-squares residual |
Output model | ![]() PDB-7kpn: |