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- PDB-5ids: Crystal Structure of a Glucose-1-phosphate Thymidylyltransferase ... -

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Basic information

Entry
Database: PDB / ID: 5ids
TitleCrystal Structure of a Glucose-1-phosphate Thymidylyltransferase from Burkholderia vietnamiensis
ComponentsGlucose-1-phosphate thymidylyltransferase
KeywordsTRANSFERASE / SSGCID / Glucose-1-phosphate thymidylyltransferase / Burkholderia vietnamiensis / Nucleotidyltransferase / Structural Genomics / Seattle Structural Genomics Center for Infectious Disease
Function / homology
Function and homology information


glucose-1-phosphate thymidylyltransferase / glucose-1-phosphate thymidylyltransferase activity / : / metal ion binding
Similarity search - Function
Glucose-1-phosphate thymidylyltransferase, short form / Nucleotidyl transferase domain / Nucleotidyl transferase / Spore Coat Polysaccharide Biosynthesis Protein SpsA; Chain A / Spore Coat Polysaccharide Biosynthesis Protein SpsA; Chain A / Nucleotide-diphospho-sugar transferases / Alpha-Beta Complex / Alpha Beta
Similarity search - Domain/homology
Glucose-1-phosphate thymidylyltransferase
Similarity search - Component
Biological speciesBurkholderia vietnamiensis (bacteria)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / molecular replacement / Resolution: 2.3 Å
AuthorsSeattle Structural Genomics Center for Infectious Disease (SSGCID)
CitationJournal: to be published
Title: Crystal Structure of a Glucose-1-phosphate Thymidylyltransferase from Burkholderia vietnamiensis
Authors: Dranow, D.M. / Hornayi, P.S. / Lorimer, D.D. / Edwards, T.E.
History
DepositionFeb 24, 2016Deposition site: RCSB / Processing site: RCSB
Revision 1.0Mar 9, 2016Provider: repository / Type: Initial release
Revision 1.1Nov 22, 2017Group: Derived calculations / Refinement description / Category: pdbx_struct_oper_list / software
Item: _pdbx_struct_oper_list.symmetry_operation / _software.name
Revision 1.2Sep 27, 2023Group: Data collection / Database references / Refinement description
Category: chem_comp_atom / chem_comp_bond ...chem_comp_atom / chem_comp_bond / database_2 / pdbx_initial_refinement_model
Item: _database_2.pdbx_DOI / _database_2.pdbx_database_accession

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: Glucose-1-phosphate thymidylyltransferase
B: Glucose-1-phosphate thymidylyltransferase
C: Glucose-1-phosphate thymidylyltransferase
D: Glucose-1-phosphate thymidylyltransferase


Theoretical massNumber of molelcules
Total (without water)133,2754
Polymers133,2754
Non-polymers00
Water3,513195
1
A: Glucose-1-phosphate thymidylyltransferase
B: Glucose-1-phosphate thymidylyltransferase


Theoretical massNumber of molelcules
Total (without water)66,6372
Polymers66,6372
Non-polymers00
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area2340 Å2
ΔGint-20 kcal/mol
Surface area22950 Å2
MethodPISA
2
C: Glucose-1-phosphate thymidylyltransferase
D: Glucose-1-phosphate thymidylyltransferase


Theoretical massNumber of molelcules
Total (without water)66,6372
Polymers66,6372
Non-polymers00
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area2520 Å2
ΔGint-17 kcal/mol
Surface area22760 Å2
MethodPISA
Unit cell
Length a, b, c (Å)112.410, 115.580, 95.810
Angle α, β, γ (deg.)90.000, 90.360, 90.000
Int Tables number5
Space group name H-MC121
Components on special symmetry positions
IDModelComponents
11B-327-

HOH

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Components

#1: Protein
Glucose-1-phosphate thymidylyltransferase


Mass: 33318.730 Da / Num. of mol.: 4 / Fragment: BuviA.00118.c.B1
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Burkholderia vietnamiensis (strain G4 / LMG 22486) (bacteria)
Strain: G4 / LMG 22486 / Gene: Bcep1808_3215 / Plasmid: BuviA.00118.c.B1 / Production host: Escherichia coli (E. coli) / Strain (production host): BL21(DE3)
References: UniProt: A4JIV2, glucose-1-phosphate thymidylyltransferase
#2: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 195 / 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: 2.33 Å3/Da / Density % sol: 47.32 %
Crystal growTemperature: 290 K / Method: vapor diffusion, sitting drop / pH: 6.5
Details: BuviA.00118.c.B1.PS02537 at 20.28 mg/ml, protein was mixed 1:1 with MCSG1(h5): 0.2 M potassium chloride, 20% (w/v) PEG-3350, cryoprotected with 20% ethylene glycol

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: APS / Beamline: 21-ID-F / Wavelength: 0.97872 Å
DetectorType: MARMOSAIC 225 mm CCD / Detector: CCD / Date: Feb 3, 2016 / Details: Beryllium Lenses
RadiationMonochromator: Diamond [111] / Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.97872 Å / Relative weight: 1
ReflectionResolution: 2.3→50 Å / Num. obs: 54334 / % possible obs: 99.9 % / Observed criterion σ(I): -3 / Redundancy: 4.3 % / Biso Wilson estimate: 41.64 Å2 / CC1/2: 0.999 / Rmerge(I) obs: 0.061 / Net I/σ(I): 15.09
Reflection shell
Resolution (Å)Highest resolution (Å)Rmerge(I) obsMean I/σ(I) obsDiffraction-ID% possible all
2.3-2.360.5522.361100
2.36-2.420.4852.63199.9
2.42-2.490.3773.37199.9
2.49-2.570.2984.3199.9
2.57-2.660.2375.431100
2.66-2.750.2016.381100
2.75-2.850.1558.2199.9
2.85-2.970.12310.28199.9
2.97-3.10.09612.93199.9
3.1-3.250.07516.111100
3.25-3.430.05920.231100
3.43-3.640.0524.281100
3.64-3.890.04427.141100
3.89-4.20.03831.611100
4.2-4.60.03733.571100
4.6-5.140.03435.131100
5.14-5.940.034341100
5.94-7.270.03533.97199.9
7.27-10.290.03240.11100
10.290.03140.63196.7

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Phasing

PhasingMethod: molecular replacement

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Processing

Software
NameVersionClassification
XSCALEdata scaling
PHENIXrefinement
PDB_EXTRACT3.2data extraction
XDSdata reduction
MoRDaphasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: 1FXO
Resolution: 2.3→48.611 Å / SU ML: 0.3 / Cross valid method: FREE R-VALUE / σ(F): 1.34 / Phase error: 25.61
RfactorNum. reflection% reflection
Rfree0.2309 2088 3.84 %
Rwork0.1761 --
obs0.1781 54309 99.87 %
Solvent computationShrinkage radii: 0.9 Å / VDW probe radii: 1.11 Å
Displacement parametersBiso max: 141.84 Å2 / Biso mean: 56.0287 Å2 / Biso min: 26.34 Å2
Refinement stepCycle: final / Resolution: 2.3→48.611 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms8477 0 0 195 8672
Biso mean---48.7 -
Num. residues----1152
Refine LS restraints
Refine-IDTypeDev idealNumber
X-RAY DIFFRACTIONf_bond_d0.0088702
X-RAY DIFFRACTIONf_angle_d0.93511902
X-RAY DIFFRACTIONf_chiral_restr0.0571337
X-RAY DIFFRACTIONf_plane_restr0.0061567
X-RAY DIFFRACTIONf_dihedral_angle_d12.3265116
LS refinement shell

Refine-ID: X-RAY DIFFRACTION / Total num. of bins used: 15

Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkNum. reflection all% reflection obs (%)
2.3001-2.35360.31531430.253134543597100
2.3536-2.41240.30531460.237434723618100
2.4124-2.47760.29771540.216834323586100
2.4776-2.55060.3071740.212534343608100
2.5506-2.63290.27011640.196334363600100
2.6329-2.7270.31471290.201835023631100
2.727-2.83610.28441490.199134573606100
2.8361-2.96520.25471380.20434783616100
2.9652-3.12150.25631170.19735093626100
3.1215-3.3170.2541000.195634903590100
3.317-3.57310.27151350.186235043639100
3.5731-3.93250.20921380.168434733611100
3.9325-4.50120.17951240.141435303654100
4.5012-5.66970.18131370.144935153652100
5.6697-48.6220.1871400.15853535367599
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)
16.5889-2.86121.14581.6706-0.43152.31990.09990.3184-0.27320.0330.07540.0990.0349-0.1521-0.20840.3202-0.06830.0840.39420.03210.309318.74123.303537.1709
21.5562-0.4151-0.25572.27470.09793.89710.068-0.17620.2033-0.00780.1801-0.3592-0.18130.2306-0.21560.2577-0.06330.01270.4442-0.08190.417928.64532.095942.9756
35.9017-3.82490.999.8697-2.63385.0121-0.0341-1.5020.33140.57040.35090.65210.8758-0.8193-0.32730.5432-0.0404-0.03130.78860.01080.442316.3574-12.028855.2041
42.9538-1.2803-0.35862.79540.38951.96260.0199-0.271-0.18720.27880.1411-0.05840.1552-0.0214-0.14470.2988-0.0683-0.05990.3910.00240.292221.1771-7.287849.302
56.3081-0.125-1.57943.7110.23273.43870.2291-0.3940.7706-0.05820.014-0.4684-0.706-0.3607-0.21820.624-0.05510.0690.462-0.21380.574525.394821.733146.9305
60.8245-1.27510.18872.7744-1.57542.0561-0.13611.05711.0765-0.1688-0.3432-0.2374-0.670.33670.21390.51410.00450.2110.71510.16860.52036.498619.774323.588
71.57330.426-0.1291.3632-0.64512.17870.15640.83930.2336-0.44230.031-0.2046-0.0772-0.1377-0.08180.54830.14420.15380.79440.12750.43614.944914.389319.0542
82.1711-0.00780.92184.3243-3.28453.68210.01750.50911.01260.22030.07320.2319-0.5007-0.39090.00250.52870.16670.10510.72840.26420.6185-9.685126.163525.3346
93.3861-0.39360.48071.9302-0.95515.6561-0.1770.40121.4345-0.32270.3768-0.1408-0.46930.0289-0.00230.76120.21670.15820.86530.42660.9108-4.25133.288317.0683
102.87640.84841.47462.6370.79862.652-0.1017-0.0742-0.09110.30570.18650.0921-0.1172-0.3818-0.10370.36590.08980.07830.45710.05230.2939-3.287214.100538.1192
112.45621.903-0.28693.4155-2.34116.84880.01140.2663-0.3301-0.5207-0.0719-0.13490.5746-0.2980.02870.4670.03750.04460.5538-0.07390.3898-4.3871-2.619324.7791
122.05521.18411.11351.06861.12062.46120.2464-0.6399-0.3653-0.04410.0350.20980.1121-0.3081-0.37930.4612-0.11140.09520.35790.01510.529632.86151.263212.3023
139.6641-5.58392.41297.4366-2.66650.9812-0.1158-0.367-0.20060.26480.1019-0.06410.0576-0.3567-0.04490.3901-0.09710.07530.3338-0.07090.292544.53169.055915.1067
142.11420.04750.17092.391-1.20952.7062-0.0648-0.03420.0430.11870.13480.3519-0.1554-0.1673-0.03160.3369-0.08170.10710.3745-0.07090.460529.772910.409613.5447
153.02943.28971.03996.3264-1.35155.279-0.1158-0.1062-0.5077-0.11610.4354-0.16480.5694-0.1125-0.34880.4618-0.10040.16160.4899-0.1170.559825.9972-1.211317.8931
163.24781.3708-0.1743.1906-1.04712.5354-0.27510.1754-0.6735-0.5370.1373-0.13410.8816-0.25160.10260.6192-0.15640.10880.4315-0.12760.567130.3741-6.79960.55
175.9052-1.62523.68420.8723-1.78757.42680.08030.11250.1048-0.15930.0266-0.30890.2459-0.2106-0.03090.377-0.08990.12860.3105-0.0760.32645.80358.24730.6434
182.39610.7043-3.26173.9554-1.67836.0996-0.15550.49070.3410.35350.19180.1614-0.5440.1117-0.10010.3773-0.03710.01350.3871-0.01930.484829.601119.25211.8063
196.6051-1.2878-4.94085.0471-1.81945.93340.5182-0.53691.12950.7144-0.2420.191-1.7820.4542-0.41850.7856-0.13340.12220.4168-0.0240.738932.758627.70346.2705
207.02993.55764.21442.83673.07554.2342-0.4368-0.50980.22410.22620.00310.2717-0.4870.06620.4670.487-0.04220.09470.3531-0.01790.438654.068922.416525.1783
213.9607-4.89360.37896.0764-0.71431.5614-0.4562-0.0922-0.64690.06020.19950.402-0.0145-0.20470.11030.465-0.13210.08010.3972-0.02790.366249.143214.087217.6031
223.96270.4167-0.82284.44810.87484.95550.1272-0.6212-0.45280.7163-0.33520.19190.4165-0.50710.30290.5421-0.14150.04030.49760.00620.359956.017713.5829.4764
234.23190.2129-1.15344.14741.25384.5885-0.0014-0.29220.1330.4149-0.1697-0.0489-0.16480.03280.06810.3919-0.085-0.05510.3665-0.0010.291963.25323.444428.5735
244.15340.426-0.41825.5630.79115.62320.0278-0.09880.6189-0.1887-0.1669-0.0308-0.56780.22520.16090.3719-0.1277-0.07320.3541-0.03120.49164.705535.500422.4304
251.13530.42140.3592.02920.88493.0662-0.0851-0.0396-0.07560.098-0.0021-0.2120.07080.25420.06410.3408-0.05730.02790.35410.00840.358164.505818.517818.7538
267.9405-6.917-0.2548.908-2.64712.87640.03990.2893-1.89620.9952-0.51260.89411.0698-0.53760.40250.7134-0.06430.03280.3759-0.05080.926161.7245-3.82921.9324
Refinement TLS group
IDRefine-IDRefine TLS-IDSelection detailsAuth asym-IDAuth seq-ID
1X-RAY DIFFRACTION1chain 'A' and (resid 2 through 35 )A2 - 35
2X-RAY DIFFRACTION2chain 'A' and (resid 36 through 139 )A36 - 139
3X-RAY DIFFRACTION3chain 'A' and (resid 140 through 156 )A140 - 156
4X-RAY DIFFRACTION4chain 'A' and (resid 157 through 246 )A157 - 246
5X-RAY DIFFRACTION5chain 'A' and (resid 247 through 290 )A247 - 290
6X-RAY DIFFRACTION6chain 'B' and (resid 3 through 19 )B3 - 19
7X-RAY DIFFRACTION7chain 'B' and (resid 20 through 104 )B20 - 104
8X-RAY DIFFRACTION8chain 'B' and (resid 105 through 182 )B105 - 182
9X-RAY DIFFRACTION9chain 'B' and (resid 183 through 218 )B183 - 218
10X-RAY DIFFRACTION10chain 'B' and (resid 219 through 247 )B219 - 247
11X-RAY DIFFRACTION11chain 'B' and (resid 248 through 290 )B248 - 290
12X-RAY DIFFRACTION12chain 'C' and (resid 2 through 17 )C2 - 17
13X-RAY DIFFRACTION13chain 'C' and (resid 18 through 35 )C18 - 35
14X-RAY DIFFRACTION14chain 'C' and (resid 36 through 76 )C36 - 76
15X-RAY DIFFRACTION15chain 'C' and (resid 77 through 100 )C77 - 100
16X-RAY DIFFRACTION16chain 'C' and (resid 101 through 218 )C101 - 218
17X-RAY DIFFRACTION17chain 'C' and (resid 219 through 246 )C219 - 246
18X-RAY DIFFRACTION18chain 'C' and (resid 247 through 265 )C247 - 265
19X-RAY DIFFRACTION19chain 'C' and (resid 266 through 290 )C266 - 290
20X-RAY DIFFRACTION20chain 'D' and (resid 2 through 19 )D2 - 19
21X-RAY DIFFRACTION21chain 'D' and (resid 20 through 35 )D20 - 35
22X-RAY DIFFRACTION22chain 'D' and (resid 36 through 76 )D36 - 76
23X-RAY DIFFRACTION23chain 'D' and (resid 77 through 139 )D77 - 139
24X-RAY DIFFRACTION24chain 'D' and (resid 140 through 199 )D140 - 199
25X-RAY DIFFRACTION25chain 'D' and (resid 200 through 265 )D200 - 265
26X-RAY DIFFRACTION26chain 'D' and (resid 266 through 290 )D266 - 290

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