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- PDB-3itk: Crystal structure of human UDP-glucose dehydrogenase Thr131Ala, a... -

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Basic information

Entry
Database: PDB / ID: 3itk
TitleCrystal structure of human UDP-glucose dehydrogenase Thr131Ala, apo form.
ComponentsUDP-glucose 6-dehydrogenase
KeywordsOXIDOREDUCTASE / Structural Genomics / Structural Genomics Consortium / SGC / NAD / Phosphoprotein
Function / homology
Function and homology information


Formation of the active cofactor, UDP-glucuronate / chondroitin sulfate biosynthetic process / UDP-glucose 6-dehydrogenase / UDP-glucose 6-dehydrogenase activity / UDP-glucuronate biosynthetic process / heparan sulfate proteoglycan biosynthetic process / glycosaminoglycan biosynthetic process / gastrulation with mouth forming second / protein hexamerization / neuron development ...Formation of the active cofactor, UDP-glucuronate / chondroitin sulfate biosynthetic process / UDP-glucose 6-dehydrogenase / UDP-glucose 6-dehydrogenase activity / UDP-glucuronate biosynthetic process / heparan sulfate proteoglycan biosynthetic process / glycosaminoglycan biosynthetic process / gastrulation with mouth forming second / protein hexamerization / neuron development / NAD binding / extracellular exosome / nucleoplasm / identical protein binding / nucleus / cytosol
Similarity search - Function
UDP-glucose 6-dehydrogenase, eukaryotic type / UDP-glucose/GDP-mannose dehydrogenase, N-terminal / UDP-glucose/GDP-mannose dehydrogenase, dimerisation / UDP-glucose/GDP-mannose dehydrogenase, C-terminal / UDP-glucose/GDP-mannose dehydrogenase / UDP-glucose/GDP-mannose dehydrogenase, C-terminal domain superfamily / UDP-glucose/GDP-mannose dehydrogenase family, central domain / UDP-glucose/GDP-mannose dehydrogenase family, UDP binding domain / UDP-glucose/GDP-mannose dehydrogenase family, NAD binding domain / UDP binding domain ...UDP-glucose 6-dehydrogenase, eukaryotic type / UDP-glucose/GDP-mannose dehydrogenase, N-terminal / UDP-glucose/GDP-mannose dehydrogenase, dimerisation / UDP-glucose/GDP-mannose dehydrogenase, C-terminal / UDP-glucose/GDP-mannose dehydrogenase / UDP-glucose/GDP-mannose dehydrogenase, C-terminal domain superfamily / UDP-glucose/GDP-mannose dehydrogenase family, central domain / UDP-glucose/GDP-mannose dehydrogenase family, UDP binding domain / UDP-glucose/GDP-mannose dehydrogenase family, NAD binding domain / UDP binding domain / Cytochrome c1, transmembrane anchor, C-terminal / 6-phosphogluconate dehydrogenase-like, C-terminal domain superfamily / Single alpha-helices involved in coiled-coils or other helix-helix interfaces / NAD(P)-binding Rossmann-like Domain / NAD(P)-binding domain superfamily / Up-down Bundle / Rossmann fold / 3-Layer(aba) Sandwich / Mainly Alpha / Alpha Beta
Similarity search - Domain/homology
UDP-glucose 6-dehydrogenase
Similarity search - Component
Biological speciesHomo sapiens (human)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 2.4 Å
AuthorsChaikuad, A. / Egger, S. / Yue, W.W. / Sethi, R. / Filippakopoulos, P. / Muniz, J.R.C. / von Delft, F. / Bountra, C. / Arrowsmith, C.H. / Weigelt, J. ...Chaikuad, A. / Egger, S. / Yue, W.W. / Sethi, R. / Filippakopoulos, P. / Muniz, J.R.C. / von Delft, F. / Bountra, C. / Arrowsmith, C.H. / Weigelt, J. / Edwards, A.M. / Kavanagh, K.L. / Nidetzky, B. / Oppermann, U. / Structural Genomics Consortium (SGC)
CitationJournal: J.Biol.Chem. / Year: 2011
Title: Structure and mechanism of human UDP-glucose 6-dehydrogenase.
Authors: Egger, S. / Chaikuad, A. / Kavanagh, K.L. / Oppermann, U. / Nidetzky, B.
History
DepositionAug 28, 2009Deposition site: RCSB / Processing site: RCSB
Revision 1.0Sep 15, 2009Provider: repository / Type: Initial release
Revision 1.1Jul 13, 2011Group: Advisory / Version format compliance
Revision 1.2Aug 31, 2011Group: Database references
Revision 1.3Oct 13, 2021Group: Database references / Derived calculations / Category: database_2 / struct_ref_seq_dif / struct_site
Item: _database_2.pdbx_DOI / _database_2.pdbx_database_accession ..._database_2.pdbx_DOI / _database_2.pdbx_database_accession / _struct_ref_seq_dif.details / _struct_site.pdbx_auth_asym_id / _struct_site.pdbx_auth_comp_id / _struct_site.pdbx_auth_seq_id
Revision 1.4Sep 6, 2023Group: Data collection / Refinement description
Category: chem_comp_atom / chem_comp_bond / pdbx_initial_refinement_model

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: UDP-glucose 6-dehydrogenase
B: UDP-glucose 6-dehydrogenase
C: UDP-glucose 6-dehydrogenase
D: UDP-glucose 6-dehydrogenase
E: UDP-glucose 6-dehydrogenase
F: UDP-glucose 6-dehydrogenase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)314,34728
Polymers312,3206
Non-polymers2,02622
Water21,4021188
1


  • Idetical with deposited unit
  • defined by author&software
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area24460 Å2
ΔGint-157 kcal/mol
Surface area103120 Å2
MethodPISA
Unit cell
Length a, b, c (Å)89.060, 106.610, 349.120
Angle α, β, γ (deg.)90.00, 90.00, 90.00
Int Tables number19
Space group name H-MP212121
Noncrystallographic symmetry (NCS)NCS domain:
IDEns-IDDetails
11A
21E
31B
41C
51D
12A
22E
32B
42C
52D
13A
23E
33B
43C
53D
63F
14A
24E
34B
44C
54D

NCS ensembles :
ID
1
2
3
4
DetailsTHE HEXAMER IN THE ASYMMETRIC UNIT REPRESENTS THE HEXAMERIC BIOLOGICAL UNIT.

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Components

#1: Protein
UDP-glucose 6-dehydrogenase / UDP-Glc dehydrogenase / UDP-GlcDH / UDPGDH


Mass: 52053.395 Da / Num. of mol.: 6 / Fragment: UNP residues 1-466 / Mutation: T131A
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Homo sapiens (human) / Gene: UGDH / Plasmid: pBEN1 / Production host: Escherichia coli (E. coli) / Strain (production host): BL21(DE3)-R3-pRARE2 / References: UniProt: O60701, UDP-glucose 6-dehydrogenase
#2: Chemical
ChemComp-PG4 / TETRAETHYLENE GLYCOL


Mass: 194.226 Da / Num. of mol.: 5 / Source method: obtained synthetically / Formula: C8H18O5 / Comment: precipitant*YM
#3: Chemical
ChemComp-EDO / 1,2-ETHANEDIOL / ETHYLENE GLYCOL


Mass: 62.068 Da / Num. of mol.: 17 / Source method: obtained synthetically / Formula: C2H6O2
#4: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 1188 / 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: 2.65 Å3/Da / Density % sol: 53.64 %
Crystal growTemperature: 293.15 K / pH: 6
Details: 15% PEG smears (PEG 400, 600, 1000, 2000, 3350, 4000, MME 5000, 6000, 8000, 10000), 0.1M MES pH 6.0, VAPOR DIFFUSION, SITTING DROP, temperature 293.15K

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: Diamond / Beamline: I03 / Wavelength: 0.9763 Å
DetectorType: ADSC QUANTUM 315 / Detector: CCD / Date: Jul 15, 2009 / Details: MIRRORS
RadiationMonochromator: SI(111) DOUBLE CRYSTAL / Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.9763 Å / Relative weight: 1
ReflectionResolution: 2.4→52.9 Å / Num. obs: 130411 / % possible obs: 99.8 % / Observed criterion σ(I): 2 / Redundancy: 4 % / Biso Wilson estimate: 40.1 Å2 / Rmerge(I) obs: 0.131 / Net I/σ(I): 7.9
Reflection shellResolution: 2.4→2.53 Å / Redundancy: 3.3 % / Rmerge(I) obs: 0.55 / Mean I/σ(I) obs: 2.1 / % possible all: 99.4

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Processing

Software
NameVersionClassification
ADSCQuantumdata collection
PHASERphasing
REFMAC5.5.0089refinement
MOSFLMdata reduction
SCALAdata scaling
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: PDB ENTRY 2Q3E
Resolution: 2.4→52.9 Å / Cor.coef. Fo:Fc: 0.935 / Cor.coef. Fo:Fc free: 0.906 / SU B: 16.607 / SU ML: 0.174 / TLS residual ADP flag: LIKELY RESIDUAL / Cross valid method: THROUGHOUT / ESU R: 0.381 / ESU R Free: 0.246 / Stereochemistry target values: MAXIMUM LIKELIHOOD
Details: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS. U VALUES: RESIDUAL ONLY. THE NCS RESTRAINTS HAVE BEEN USED IN REFINEMENT.
RfactorNum. reflection% reflectionSelection details
Rfree0.241 2134 1.6 %RANDOM
Rwork0.198 ---
obs0.199 128277 99.7 %-
all-130411 --
Solvent computationIon probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.4 Å / Solvent model: MASK
Displacement parametersBiso mean: 12.77 Å2
Baniso -1Baniso -2Baniso -3
1--0.23 Å20 Å20 Å2
2--0.37 Å20 Å2
3----0.14 Å2
Refine analyzeLuzzati coordinate error obs: 0.328 Å
Refinement stepCycle: LAST / Resolution: 2.4→52.9 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms21621 0 133 1188 22942
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.0160.02222169
X-RAY DIFFRACTIONr_bond_other_d0.0030.0215064
X-RAY DIFFRACTIONr_angle_refined_deg1.4631.9629990
X-RAY DIFFRACTIONr_angle_other_deg1.096336789
X-RAY DIFFRACTIONr_dihedral_angle_1_deg5.44952795
X-RAY DIFFRACTIONr_dihedral_angle_2_deg35.48724.344976
X-RAY DIFFRACTIONr_dihedral_angle_3_deg13.546153816
X-RAY DIFFRACTIONr_dihedral_angle_4_deg17.50115142
X-RAY DIFFRACTIONr_chiral_restr0.0830.23425
X-RAY DIFFRACTIONr_gen_planes_refined0.0070.02124613
X-RAY DIFFRACTIONr_gen_planes_other0.0030.024367
X-RAY DIFFRACTIONr_nbd_refined
X-RAY DIFFRACTIONr_nbd_other
X-RAY DIFFRACTIONr_nbtor_refined
X-RAY DIFFRACTIONr_nbtor_other
X-RAY DIFFRACTIONr_xyhbond_nbd_refined
X-RAY DIFFRACTIONr_xyhbond_nbd_other
X-RAY DIFFRACTIONr_metal_ion_refined
X-RAY DIFFRACTIONr_metal_ion_other
X-RAY DIFFRACTIONr_symmetry_vdw_refined
X-RAY DIFFRACTIONr_symmetry_vdw_other
X-RAY DIFFRACTIONr_symmetry_hbond_refined
X-RAY DIFFRACTIONr_symmetry_hbond_other
X-RAY DIFFRACTIONr_symmetry_metal_ion_refined
X-RAY DIFFRACTIONr_symmetry_metal_ion_other
X-RAY DIFFRACTIONr_mcbond_it1.793313907
X-RAY DIFFRACTIONr_mcbond_other0.61835650
X-RAY DIFFRACTIONr_mcangle_it3.257522479
X-RAY DIFFRACTIONr_scbond_it5.50688262
X-RAY DIFFRACTIONr_scangle_it7.877117506
X-RAY DIFFRACTIONr_rigid_bond_restr
X-RAY DIFFRACTIONr_sphericity_free
X-RAY DIFFRACTIONr_sphericity_bonded
Refine LS restraints NCS

Refine-ID: X-RAY DIFFRACTION

Ens-IDDom-IDAuth asym-IDNumberTypeRms dev position (Å)Weight position
11A457tight positional0.040.05
11B457tight positional0.030.05
11C457tight positional0.040.05
11D457tight positional0.040.05
11E457tight positional0.050.05
22A847tight positional0.040.05
22B847tight positional0.050.05
22C847tight positional0.060.05
22D847tight positional0.050.05
22E847tight positional0.070.05
33A267tight positional0.040.05
33B267tight positional0.060.05
33C267tight positional0.040.05
33D267tight positional0.040.05
33E267tight positional0.040.05
33F267tight positional0.050.05
44A895tight positional0.130.05
44B895tight positional0.140.05
44C895tight positional0.150.05
44D895tight positional0.10.05
44E895tight positional0.120.05
11A523medium positional0.060.5
11B523medium positional0.050.5
11C523medium positional0.130.5
11D523medium positional0.050.5
11E523medium positional0.060.5
22A917medium positional0.050.5
22B917medium positional0.090.5
22C917medium positional0.060.5
22D917medium positional0.050.5
22E917medium positional0.10.5
33A271medium positional0.050.5
33B271medium positional0.070.5
33C271medium positional0.050.5
33D271medium positional0.140.5
33E271medium positional0.050.5
33F271medium positional0.060.5
44A1610medium positional0.220.5
44B1610medium positional0.30.5
44C1610medium positional0.320.5
44D1610medium positional0.180.5
44E1610medium positional0.250.5
11A457tight thermal0.110.5
11B457tight thermal0.110.5
11C457tight thermal0.140.5
11D457tight thermal0.130.5
11E457tight thermal0.140.5
22A847tight thermal0.140.5
22B847tight thermal0.160.5
22C847tight thermal0.160.5
22D847tight thermal0.140.5
22E847tight thermal0.160.5
33A267tight thermal0.170.5
33B267tight thermal0.180.5
33C267tight thermal0.160.5
33D267tight thermal0.190.5
33E267tight thermal0.160.5
33F267tight thermal0.190.5
44A895tight thermal0.990.5
44B895tight thermal0.930.5
44C895tight thermal1.030.5
44D895tight thermal0.940.5
44E895tight thermal0.880.5
11A523medium thermal0.122
11B523medium thermal0.092
11C523medium thermal0.122
11D523medium thermal0.112
11E523medium thermal0.112
22A917medium thermal0.122
22B917medium thermal0.132
22C917medium thermal0.142
22D917medium thermal0.122
22E917medium thermal0.132
33A271medium thermal0.152
33B271medium thermal0.152
33C271medium thermal0.142
33D271medium thermal0.142
33E271medium thermal0.142
33F271medium thermal0.172
44A1610medium thermal1.042
44B1610medium thermal0.962
44C1610medium thermal1.082
44D1610medium thermal1.022
44E1610medium thermal1.062
LS refinement shellResolution: 2.4→2.46 Å / Total num. of bins used: 20
RfactorNum. reflection% reflection
Rfree0.355 129 -
Rwork0.318 9292 -
obs--98.55 %
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)
10.3789-0.2314-0.2170.6521-0.34953.435-0.0063-0.2842-0.0799-0.02590.0608-0.00890.05390.5257-0.05450.1837-0.0342-0.05580.30740.04510.078822.7497-16.09827.8955
20.5348-0.1017-0.15782.93540.53792.9233-0.0511-0.1438-0.13050.01710.12950.04930.0275-0.1211-0.07840.2019-0.0568-0.07380.19030.1120.14910.028-16.12093.5163
31.06660.22830.18120.3481-0.10450.32270.0909-0.0337-0.11530.04720.00220.0120.0641-0.0166-0.09310.16260.0048-0.05170.0497-0.02740.058929.42059.4161-13.1758
41.13030.1188-0.27071.4157-0.41881.9866-0.00220.02550.1443-0.0556-0.1985-0.1856-0.04910.32070.20070.0932-0.0109-0.01980.09970.0430.170461.097224.0955-40.9747
51.48750.4315-0.01460.5676-0.27445.0355-0.01440.06950.2828-0.0028-0.1123-0.035-0.194-0.10020.12670.1520.0195-0.06280.0667-0.00110.199550.030331.8466-42.4552
60.593-0.37330.04061.48920.04860.0238-0.0149-0.00240.0212-0.02120.0087-0.1877-0.0397-0.02260.00620.11020.021-0.00790.12220.00550.05946.4944-0.0918-57.8219
70.48350.11870.34120.9093-0.0880.4477-0.04810.09060.12280.08180.02430.18820.0597-0.13520.02380.1559-0.0666-0.00170.30310.06940.072715.002415.1065-86.4272
81.39670.74681.42072.5285-0.1351.88580.0852-0.0122-0.02710.0624-0.1285-0.16140.1489-0.07350.04330.1038-0.04410.03470.28270.01530.027126.09137.3263-85.8254
90.7945-0.6142-0.59950.82070.44110.47680.04720.09520.0463-0.0572-0.0130.0952-0.0414-0.0315-0.03420.14190.0172-0.03040.11410.03630.102218.252525.9738-55.6078
102.80980.5352-0.43281.7933-1.57533.54810.157-0.2133-0.56060.0785-0.1774-0.19740.30830.1480.02040.17120.0148-0.02970.066-0.02520.387942.2797-51.3417-57.6441
111.2503-0.2002-0.12330.42130.09590.58050.0870.0586-0.29050.021-0.06010.03950.1232-0.0124-0.02690.10560.01920.01750.0946-0.05610.151136.7212-36.9932-63.6767
120.102-0.11930.04670.3254-0.27941.8145-0.02480.0237-0.06890.1269-0.0061-0.07920.1910.07730.03080.2083-0.0138-0.05180.13040.02160.262721.9177-34.81-32.4917
130.6378-0.3063-0.37150.89860.32091.17490.08610.01710.1181-0.1864-0.1240.1347-0.0861-0.22740.0380.05840.0242-0.0140.1197-0.07520.1173-6.725935.8581-21.1369
141.2520.2566-0.61791.4782-1.53032.9431-0.0068-0.11960.1134-0.1544-0.04140.00250.0598-0.08060.04810.08890.0050.01410.0412-0.05510.12986.206235.1769-17.0765
151.2687-0.60350.45690.87680.04830.85360.185-0.1092-0.40770.05450.03710.32590.2014-0.3132-0.22210.1691-0.0704-0.0550.17370.09120.2321-6.60931.5998-21.3521
162.4104-0.26990.59971.2052-0.82382.39110.0273-0.2035-0.0120.2406-0.07120.0726-0.0483-0.40330.04390.0637-0.05540.01640.1353-0.02150.0112-20.8269-31.2527-50.152
171.34230.1138-0.74910.1574-0.46442.70720.1087-0.3495-0.12090.1322-0.0942-0.034-0.04810.0499-0.01460.2535-0.1472-0.02750.26980.07170.0899-14.2185-36.7565-39.8411
181.03960.40560.2780.5311-0.2071.0321-0.09920.07750.040.03340.01910.0201-0.0265-0.03450.08010.0542-0.02460.0070.0972-0.01630.093.8638-20.727-62.914
Refinement TLS group
IDRefine-IDRefine TLS-IDAuth asym-IDAuth seq-ID
1X-RAY DIFFRACTION1A0 - 120
2X-RAY DIFFRACTION2A121 - 204
3X-RAY DIFFRACTION3A205 - 466
4X-RAY DIFFRACTION4C1 - 120
5X-RAY DIFFRACTION5C121 - 204
6X-RAY DIFFRACTION6C205 - 466
7X-RAY DIFFRACTION7D0 - 117
8X-RAY DIFFRACTION8D118 - 209
9X-RAY DIFFRACTION9D210 - 466
10X-RAY DIFFRACTION10E1 - 79
11X-RAY DIFFRACTION11E80 - 204
12X-RAY DIFFRACTION12E205 - 466
13X-RAY DIFFRACTION13B1 - 120
14X-RAY DIFFRACTION14B121 - 204
15X-RAY DIFFRACTION15B205 - 466
16X-RAY DIFFRACTION16F0 - 120
17X-RAY DIFFRACTION17F121 - 204
18X-RAY DIFFRACTION18F205 - 466

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