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- PDB-5yja: Crystal structure of highly active BTUO mutant P287G without dehy... -

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Basic information

Entry
Database: PDB / ID: 5yja
TitleCrystal structure of highly active BTUO mutant P287G without dehydration
ComponentsUric acid degradation bifunctional protein
KeywordsOXIDOREDUCTASE / protein engineering / enzyme / loop flexibility / entropy of activation
Function / homology
Function and homology information


2-oxo-4-hydroxy-4-carboxy-5-ureidoimidazoline decarboxylase activity / 2-oxo-4-hydroxy-4-carboxy-5-ureidoimidazoline decarboxylase / factor-independent urate hydroxylase / urate oxidase activity / allantoin metabolic process / urate catabolic process / purine nucleobase metabolic process / peroxisome
Similarity search - Function
2-oxo-4-hydroxy-4-carboxy-5-ureidoimidazoline decarboxylase, type 1 / Oxo-4-hydroxy-4-carboxy-5-ureidoimidazoline decarboxylase / Oxo-4-hydroxy-4-carboxy-5-ureidoimidazoline decarboxylase superfamily / OHCU decarboxylase / Urate Oxidase / Urate Oxidase; / Uricase, conserved site / Uricase signature. / Uricase / Uricase ...2-oxo-4-hydroxy-4-carboxy-5-ureidoimidazoline decarboxylase, type 1 / Oxo-4-hydroxy-4-carboxy-5-ureidoimidazoline decarboxylase / Oxo-4-hydroxy-4-carboxy-5-ureidoimidazoline decarboxylase superfamily / OHCU decarboxylase / Urate Oxidase / Urate Oxidase; / Uricase, conserved site / Uricase signature. / Uricase / Uricase / Roll / Alpha Beta
Similarity search - Domain/homology
8-AZAXANTHINE / : / 2-METHOXYETHANOL / OXYGEN MOLECULE / Uric acid degradation bifunctional protein
Similarity search - Component
Biological speciesBacillus sp. (bacteria)
MethodX-RAY DIFFRACTION / SYNCHROTRON / Resolution: 1.65 Å
AuthorsHibi, T. / Itoh, T. / Nishiya, Y.
Citation
Journal: to be published
Title: Flexibility of a Distal Interface Loop Modulates Water Network in the Active Site of Bacillus sp. TB-90 Urate Oxidase
Authors: Hibi, T. / Itoh, T. / Nishiya, Y.
#1: Journal: Biochemistry / Year: 2016
Title: Hyperstabilization of Tetrameric Bacillus sp. TB-90 Urate Oxidase by Introducing Disulfide Bonds through Structural Plasticity.
Authors: Hibi, T. / Kume, A. / Kawamura, A. / Itoh, T. / Fukada, H. / Nishiya, Y.
#2: Journal: Biochemistry / Year: 2014
Title: Intersubunit salt bridges with a sulfate anion control subunit dissociation and thermal stabilization of Bacillus sp. TB-90 urate oxidase.
Authors: Hibi, T. / Hayashi, Y. / Fukada, H. / Itoh, T. / Nago, T. / Nishiya, Y.
History
DepositionOct 9, 2017Deposition site: PDBJ / Processing site: PDBJ
Revision 1.0Oct 10, 2018Provider: repository / Type: Initial release
Revision 1.1Dec 19, 2018Group: Data collection / Experimental preparation / Structure summary
Category: entity / exptl_crystal_grow
Item: _entity.formula_weight / _exptl_crystal_grow.pdbx_details

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

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Assembly

Deposited unit
A: Uric acid degradation bifunctional protein
B: Uric acid degradation bifunctional protein
C: Uric acid degradation bifunctional protein
D: Uric acid degradation bifunctional protein
hetero molecules


Theoretical massNumber of molelcules
Total (without water)149,43318
Polymers148,2704
Non-polymers1,16314
Water19,4381079
1


  • Idetical with deposited unit
  • defined by author&software
  • Evidence: gel filtration
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area25680 Å2
ΔGint-99 kcal/mol
Surface area40670 Å2
MethodPISA
Unit cell
Length a, b, c (Å)133.017, 144.252, 70.272
Angle α, β, γ (deg.)90.00, 90.00, 90.00
Int Tables number18
Space group name H-MP21212

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Components

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Protein , 1 types, 4 molecules ABCD

#1: Protein
Uric acid degradation bifunctional protein


Mass: 37067.594 Da / Num. of mol.: 4 / Fragment: UNP RESIDUES 172-494 / Mutation: deletion of 1-7 and 318-331
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Bacillus sp. (strain TB-90) (bacteria) / Strain: TB-90 / Gene: uao / Production host: Escherichia coli DH5[alpha] (bacteria)
References: UniProt: Q45697, 2-oxo-4-hydroxy-4-carboxy-5-ureidoimidazoline decarboxylase, factor-independent urate hydroxylase

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Non-polymers , 6 types, 1093 molecules

#2: Chemical
ChemComp-AZA / 8-AZAXANTHINE


Mass: 153.099 Da / Num. of mol.: 4 / Source method: obtained synthetically / Formula: C4H3N5O2 / Feature type: SUBJECT OF INVESTIGATION
#3: Chemical
ChemComp-OXY / OXYGEN MOLECULE


Mass: 31.999 Da / Num. of mol.: 4 / Source method: obtained synthetically / Formula: O2 / Feature type: SUBJECT OF INVESTIGATION
#4: Chemical ChemComp-SO4 / SULFATE ION


Mass: 96.063 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: SO4
#5: Chemical ChemComp-K / POTASSIUM ION


Mass: 39.098 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: K
#6: Chemical ChemComp-MXE / 2-METHOXYETHANOL


Mass: 76.094 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: C3H8O2
#7: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 1079 / Source method: isolated from a natural source / Formula: H2O

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Experimental details

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Experiment

ExperimentMethod: X-RAY DIFFRACTION / Number of used crystals: 1

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Sample preparation

CrystalDensity Matthews: 2.26 Å3/Da / Density % sol: 45.08 %
Crystal growTemperature: 288 K / Method: vapor diffusion, hanging drop / pH: 8.5
Details: 16% PEG 8000, 0.1 M Tris-HCl, 0.07 M K2SO4, 2 mM 8-azaxanthine

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: SPring-8 / Beamline: BL38B1 / Wavelength: 1 Å
DetectorType: RAYONIX MX-225 / Detector: CCD / Date: Nov 18, 2015
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 1 Å / Relative weight: 1
ReflectionResolution: 1.65→50 Å / Num. obs: 162624 / % possible obs: 99 % / Redundancy: 11.4 % / Biso Wilson estimate: 26.9 Å2 / Rmerge(I) obs: 0.089 / Rpim(I) all: 0.027 / Rrim(I) all: 0.093 / Χ2: 1.041 / Net I/σ(I): 22.4
Reflection shellResolution: 1.65→1.68 Å / Redundancy: 10 % / Rmerge(I) obs: 0.888 / Num. unique obs: 8061 / CC1/2: 0.8 / Rpim(I) all: 0.285 / Rrim(I) all: 0.934 / Χ2: 1.032 / % possible all: 99.8

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Processing

Software
NameVersionClassification
PHENIX(1.12_2829)refinement
HKL-2000data reduction
HKL-2000data scaling
PHASERphasing
RefinementResolution: 1.65→39.683 Å / SU ML: 0.15 / Cross valid method: FREE R-VALUE / σ(F): 1.34 / Phase error: 18.36
RfactorNum. reflection% reflection
Rfree0.1867 16293 10.02 %
Rwork0.1637 --
obs0.1659 161351 99.8 %
Solvent computationShrinkage radii: 0.9 Å / VDW probe radii: 1.11 Å
Refine analyzeLuzzati coordinate error obs: 0.172 Å
Refinement stepCycle: LAST / Resolution: 1.65→39.683 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms9414 0 74 1079 10567
Refine LS restraints
Refine-IDTypeDev idealNumber
X-RAY DIFFRACTIONf_bond_d0.0099812
X-RAY DIFFRACTIONf_angle_d0.94413343
X-RAY DIFFRACTIONf_dihedral_angle_d9.0845730
X-RAY DIFFRACTIONf_chiral_restr0.0591490
X-RAY DIFFRACTIONf_plane_restr0.0071703
LS refinement shell
Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkRefine-ID% reflection obs (%)
1.6492-1.6680.25335120.22634830X-RAY DIFFRACTION99
1.668-1.68760.23695390.20774803X-RAY DIFFRACTION100
1.6876-1.70820.25075600.21044857X-RAY DIFFRACTION100
1.7082-1.72980.23665350.19984787X-RAY DIFFRACTION100
1.7298-1.75260.23545480.19534844X-RAY DIFFRACTION100
1.7526-1.77660.22355590.18524805X-RAY DIFFRACTION100
1.7766-1.8020.2155390.17924867X-RAY DIFFRACTION100
1.802-1.82890.20665510.18174819X-RAY DIFFRACTION100
1.8289-1.85740.21135420.17954847X-RAY DIFFRACTION100
1.8574-1.88790.20985090.18134857X-RAY DIFFRACTION100
1.8879-1.92040.20735590.17294825X-RAY DIFFRACTION100
1.9204-1.95540.19795350.16694894X-RAY DIFFRACTION100
1.9554-1.9930.1945300.16964829X-RAY DIFFRACTION100
1.993-2.03360.2015420.16854858X-RAY DIFFRACTION100
2.0336-2.07790.18025230.15464887X-RAY DIFFRACTION100
2.0779-2.12620.1785520.15264849X-RAY DIFFRACTION100
2.1262-2.17940.17045650.14774833X-RAY DIFFRACTION100
2.1794-2.23830.1635210.14974879X-RAY DIFFRACTION100
2.2383-2.30410.16986140.14774790X-RAY DIFFRACTION100
2.3041-2.37850.17955690.15164832X-RAY DIFFRACTION100
2.3785-2.46350.18735450.15154902X-RAY DIFFRACTION100
2.4635-2.56210.17585280.15054882X-RAY DIFFRACTION100
2.5621-2.67870.18335430.14894924X-RAY DIFFRACTION100
2.6787-2.81990.18445060.15664928X-RAY DIFFRACTION100
2.8199-2.99650.20065190.16574923X-RAY DIFFRACTION100
2.9965-3.22780.18155950.16344884X-RAY DIFFRACTION100
3.2278-3.55240.18355330.15764955X-RAY DIFFRACTION100
3.5524-4.0660.16375770.15394937X-RAY DIFFRACTION100
4.066-5.1210.14895130.13555052X-RAY DIFFRACTION100
5.121-39.69470.21755300.21055152X-RAY DIFFRACTION98
Refinement TLS params.

Method: refined / Refine-ID: X-RAY DIFFRACTION

IDL112)L122)L132)L222)L232)L332)S11 (Å °)S12 (Å °)S13 (Å °)S21 (Å °)S22 (Å °)S23 (Å °)S31 (Å °)S32 (Å °)S33 (Å °)T112)T122)T132)T222)T232)T332)Origin x (Å)Origin y (Å)Origin z (Å)
11.31160.2867-0.14361.35970.0780.2153-0.0343-0.0746-0.33680.1504-0.03190.00030.14330.02530.07240.20320.0477-0.01740.17250.03250.229115.9612163.599426.4308
20.78810.0397-0.13662.09030.12740.6432-0.0112-0.1487-0.02690.15260.0095-0.22490.07240.12720.00390.12650.0331-0.03890.19710.01290.157725.3996177.781530.51
31.1947-0.1873-0.28180.6499-0.14410.2511-0.0131-0.0586-0.15580.021-0.06450.02160.1364-0.05170.05810.1559-0.01-0.02910.1150.03230.15045.1877166.846828.558
41.3142-0.1638-0.26781.2044-0.16520.6299-0.0466-0.1655-0.24460.11450.00310.07560.1062-0.01090.0510.2268-0.0056-0.00840.1660.0360.1903-2.7792163.682535.2913
52.1203-0.48040.87120.9628-0.78242.1495-0.1052-0.2229-0.1060.1490.0834-0.02010.1714-0.0785-0.06180.20240.0014-0.0040.16920.00970.1421-9.609173.52132.4451
60.72350.3602-0.08581.4864-0.05190.53750.0397-0.0976-0.04240.2258-0.02220.16110.0972-0.0887-0.02420.1212-0.02390.02050.10260.00480.0857-10.1225191.489839.1498
71.23270.1546-0.09120.70060.18530.5191-0.0135-0.010.08730.0039-0.03670.03440.02130.02780.06190.1031-0.0021-0.00150.0677-0.00950.07030.3954198.411829.0057
81.2986-0.7635-0.36761.40760.22090.44080.0303-0.07070.0610.04060.0222-0.116-0.00410.1617-0.03020.09910.0047-0.01860.1135-0.01140.084114.0384200.197932.2014
90.9532-0.0354-0.10370.7251-0.2910.4981-0.00520.04260.04950.00310.0387-0.17690.12240.40170.02470.14870.0307-0.01530.1914-0.00380.168919.5292192.371220.2371
100.9803-0.5961-0.0091.3566-0.14410.54310.0393-0.02580.0894-0.10670.0018-0.10260.03860.0533-0.03210.0831-0.00130.00690.06500.05887.0515201.73151.66
111.3339-0.1135-0.09561.4564-0.14650.550.03920.1784-0.1307-0.2725-0.0347-0.17880.20850.0744-0.01240.18230.04750.02220.1444-0.0170.113113.32187.0245-6.9998
121.2914-0.13910.25860.76920.2132.5295-0.0694-0.02270.1606-0.0746-0.0292-0.0876-0.25380.0140.06360.1030.0096-0.00120.05990.02910.08957.8594200.05956.781
130.68740.00310.05170.68170.05340.00420.03760.0579-0.026-0.0355-0.01460.15260.0582-0.1575-0.00470.1132-0.0196-0.0210.16070.00150.1061-15.8779192.75665.4146
141.20130.7126-0.5471.4914-0.33880.99590.020.04350.0222-0.04250.01310.08390.0477-0.1668-0.02820.0894-0.0057-0.01730.10310.01060.082-12.0569200.67562.3235
150.91490.7102-0.5091.682-0.9942.69790.0147-0.00690.0874-0.02650.05120.12860.1468-0.3976-0.14590.119-0.0286-0.02130.18290.00910.1622-17.5559191.275611.7437
161.4413-0.34350.07541.4048-0.0980.4430.01090.0667-0.3722-0.1146-0.00290.02070.2084-0.0109-0.00070.2274-0.0492-0.02110.1759-0.0360.2464-13.5126162.41898.908
170.9202-0.0387-0.2121.38690.01680.4579-0.00620.1349-0.0506-0.0960.00840.2250.0532-0.07150.00490.1189-0.0389-0.03120.1888-0.01260.1632-23.4954176.56154.6683
181.9229-0.07540.7030.53160.13851.67230.0310.0434-0.17470.0506-0.11340.11370.33630.09770.02880.1755-0.0267-0.00730.0708-0.05690.1772-11.0835163.699612.4627
190.6781-0.05-0.3470.8745-0.02480.29760.06060.1642-0.0359-0.1174-0.0651-0.16830.07270.06980.02160.18560.0242-0.01060.2075-0.00950.1727.7675170.6162-1.6189
201.19640.6554-0.49651.5826-0.18250.51160.03330.0045-0.1813-0.0233-0.1004-0.31860.11930.09930.07490.24040.0183-0.00970.204-0.0140.206211.1686169.25320.8917
211.86060.2710.05011.5871-0.05221.35290.0660.2702-0.4411-0.1471-0.090.08940.1887-0.09040.00520.23940.0184-0.01850.173-0.09270.2534-1.9039158.2195-0.2002
223.16281.70751.51711.94331.31412.0103-0.15110.4499-0.3466-0.23030.1964-0.1707-0.01230.2409-0.0390.20960.00140.01590.217-0.03260.19167.5345165.4935-1.0568
231.99011.0470.71882.11150.90181.6222-0.28180.3033-0.1594-0.34910.18180.0631-0.0796-0.04790.10450.1892-0.03280.01080.1904-0.01590.16149.9886174.00071.1427
Refinement TLS group
IDRefine-IDRefine TLS-IDSelection details
1X-RAY DIFFRACTION1chain 'A' and (resid 8 through 43 )
2X-RAY DIFFRACTION2chain 'A' and (resid 44 through 119 )
3X-RAY DIFFRACTION3chain 'A' and (resid 120 through 192 )
4X-RAY DIFFRACTION4chain 'A' and (resid 193 through 291 )
5X-RAY DIFFRACTION5chain 'A' and (resid 292 through 310 )
6X-RAY DIFFRACTION6chain 'B' and (resid 9 through 119 )
7X-RAY DIFFRACTION7chain 'B' and (resid 120 through 188 )
8X-RAY DIFFRACTION8chain 'B' and (resid 189 through 276 )
9X-RAY DIFFRACTION9chain 'B' and (resid 277 through 313 )
10X-RAY DIFFRACTION10chain 'C' and (resid 9 through 43 )
11X-RAY DIFFRACTION11chain 'C' and (resid 44 through 119 )
12X-RAY DIFFRACTION12chain 'C' and (resid 120 through 161 )
13X-RAY DIFFRACTION13chain 'C' and (resid 162 through 192 )
14X-RAY DIFFRACTION14chain 'C' and (resid 193 through 276 )
15X-RAY DIFFRACTION15chain 'C' and (resid 277 through 314 )
16X-RAY DIFFRACTION16chain 'D' and (resid 8 through 43 )
17X-RAY DIFFRACTION17chain 'D' and (resid 44 through 119 )
18X-RAY DIFFRACTION18chain 'D' and (resid 120 through 161 )
19X-RAY DIFFRACTION19chain 'D' and (resid 162 through 192 )
20X-RAY DIFFRACTION20chain 'D' and (resid 193 through 213 )
21X-RAY DIFFRACTION21chain 'D' and (resid 214 through 248 )
22X-RAY DIFFRACTION22chain 'D' and (resid 249 through 293 )
23X-RAY DIFFRACTION23chain 'D' and (resid 294 through 310 )

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