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Yorodumi- PDB-3thp: Crystal structure and RNA binding properties of the RRM/AlkB doma... -
+Open data
-Basic information
Entry | Database: PDB / ID: 3thp | ||||||
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Title | Crystal structure and RNA binding properties of the RRM/AlkB domains in human ABH8, an enzyme catalyzing tRNA hypermodification, Northeast Structural Genomics Consortium Target HR5601B | ||||||
Components | Alkylated DNA repair protein alkB homolog 8 | ||||||
Keywords | OXIDOREDUCTASE / Structural Genomics / PSI-Biology / Northeast Structural Genomics Consortium / NESG / alpha-beta two domain-protein containing a zinc structure motif / tRNA modifying enzyme | ||||||
Function / homology | Function and homology information tRNA (carboxymethyluridine34-5-O)-methyltransferase / tRNA (5-carboxymethyluridine(34)-5-O)-methyltransferase activity / tRNA (uridine) methyltransferase activity / tRNA wobble uridine modification / tRNA modification in the nucleus and cytosol / 2-oxoglutarate-dependent dioxygenase activity / S-adenosylmethionine-dependent methyltransferase activity / tRNA methylation / tRNA binding / iron ion binding ...tRNA (carboxymethyluridine34-5-O)-methyltransferase / tRNA (5-carboxymethyluridine(34)-5-O)-methyltransferase activity / tRNA (uridine) methyltransferase activity / tRNA wobble uridine modification / tRNA modification in the nucleus and cytosol / 2-oxoglutarate-dependent dioxygenase activity / S-adenosylmethionine-dependent methyltransferase activity / tRNA methylation / tRNA binding / iron ion binding / DNA damage response / zinc ion binding / nucleoplasm / nucleus / cytoplasm / cytosol Similarity search - Function | ||||||
Biological species | Homo sapiens (human) | ||||||
Method | X-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 3.2 Å | ||||||
Authors | Pastore, C. / Topalidou, I. / Forouhar, F. / Yan, A.C. / Levy, M. / Hunt, J.F. / Northeast Structural Genomics Consortium (NESG) | ||||||
Citation | Journal: J.Biol.Chem. / Year: 2012 Title: Crystal structure and RNA binding properties of the RNA recognition motif (RRM) and AlkB domains in human AlkB homolog 8 (ABH8), an enzyme catalyzing tRNA hypermodification. Authors: Pastore, C. / Topalidou, I. / Forouhar, F. / Yan, A.C. / Levy, M. / Hunt, J.F. | ||||||
History |
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-Structure visualization
Structure viewer | Molecule: MolmilJmol/JSmol |
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-Downloads & links
-Download
PDBx/mmCIF format | 3thp.cif.gz | 76.3 KB | Display | PDBx/mmCIF format |
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PDB format | pdb3thp.ent.gz | 55.3 KB | Display | PDB format |
PDBx/mmJSON format | 3thp.json.gz | Tree view | PDBx/mmJSON format | |
Others | Other downloads |
-Validation report
Summary document | 3thp_validation.pdf.gz | 445.9 KB | Display | wwPDB validaton report |
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Full document | 3thp_full_validation.pdf.gz | 454.4 KB | Display | |
Data in XML | 3thp_validation.xml.gz | 14.2 KB | Display | |
Data in CIF | 3thp_validation.cif.gz | 18 KB | Display | |
Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/th/3thp ftp://data.pdbj.org/pub/pdb/validation_reports/th/3thp | HTTPS FTP |
-Related structure data
Related structure data | 3thtC 2cq2S 2fdiS C: citing same article (ref.) S: Starting model for refinement |
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Similar structure data | |
Other databases |
-Links
-Assembly
Deposited unit |
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Unit cell |
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Details | The light scattering data of this construct without C-tag shows mainly monomeric protein in the solution, while the same construct with C-tag appears to have population of monomer and tetrameric species. Consistent with the latter observation, the crystal structure reveals that a few residues of the C-tag play an important role in protein tetramerization, which is possibly not a physiological phenomenon |
-Components
#1: Protein | Mass: 39687.945 Da / Num. of mol.: 1 / Fragment: RRM and AlkB domains of ABH8 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: ABH8, ALKBH8 / Production host: Escherichia coli (E. coli) / Strain (production host): B834(lambda)(DE3) References: UniProt: Q96BT7, Oxidoreductases; Acting on paired donors, with incorporation or reduction of molecular oxygen; With 2-oxoglutarate as one donor, and incorporation of one atom of oxygen into each donor |
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#2: Chemical | ChemComp-ZN / |
#3: Chemical | ChemComp-MN / |
#4: Chemical | ChemComp-AKG / |
-Experimental details
-Experiment
Experiment | Method: X-RAY DIFFRACTION / Number of used crystals: 1 |
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-Sample preparation
Crystal | Density Matthews: 2.54 Å3/Da / Density % sol: 51.61 % |
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Crystal grow | Temperature: 293 K / Method: microbatch under oil / pH: 7 Details: 30% PEG 3350 and 0.15 M D/L malic acid, 2.8 mM MnCl2, and 8.6 mM 2-oxyglutarate., pH 7, microbatch under oil, temperature 293K |
-Data collection
Diffraction | Mean temperature: 100 K |
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Diffraction source | Source: SYNCHROTRON / Site: NSLS / Beamline: X4C / Wavelength: 0.97885 Å |
Detector | Type: MAR CCD 165 mm / Detector: CCD / Date: Nov 10, 2010 / Details: mirrors |
Radiation | Monochromator: Si 111 CHANNEL / Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray |
Radiation wavelength | Wavelength: 0.97885 Å / Relative weight: 1 |
Reflection | Resolution: 3.2→40 Å / Num. all: 12782 / Num. obs: 12770 / % possible obs: 99.9 % / Observed criterion σ(F): 0 / Observed criterion σ(I): 0 / Redundancy: 7.4 % / Biso Wilson estimate: 42.5 Å2 / Rmerge(I) obs: 0.154 / Rsym value: 0.124 / Net I/σ(I): 14.3 |
Reflection shell | Resolution: 3.2→3.31 Å / Redundancy: 6.4 % / Rmerge(I) obs: 0.437 / Mean I/σ(I) obs: 3.7 / Num. unique all: 1293 / Rsym value: 0.393 / % possible all: 99.6 |
-Processing
Software |
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Refinement | Method to determine structure: MOLECULAR REPLACEMENT Starting model: PDB entry 2CQ2 for RRM domain, PDB entry 2FDI for AlkB domain. Resolution: 3.2→35.57 Å / Rfactor Rfree error: 0.008 / Data cutoff high absF: 296587.36 / Data cutoff low absF: 0 / Isotropic thermal model: RESTRAINED / Cross valid method: THROUGHOUT / σ(F): 0 / σ(I): 0 / Stereochemistry target values: Engh & Huber
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Solvent computation | Solvent model: FLAT MODEL / Bsol: 24.031 Å2 / ksol: 0.35 e/Å3 | |||||||||||||||||||||||||
Displacement parameters | Biso mean: 44.2 Å2
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Refine analyze |
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Refinement step | Cycle: LAST / Resolution: 3.2→35.57 Å
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Refine LS restraints |
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LS refinement shell | Resolution: 3.2→3.31 Å / Rfactor Rfree error: 0.041 / Total num. of bins used: 10
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