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- PDB-4wgh: Crystal structure of aldo/keto reductase from Klebsiella pneumoni... -

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Basic information

Entry
Database: PDB / ID: 4wgh
TitleCrystal structure of aldo/keto reductase from Klebsiella pneumoniae in complex with NADP and acetate at 1.8 A resolution
ComponentsAldehyde reductase
KeywordsOXIDOREDUCTASE / Aldo/keto reductase / NADP / Klebsiella pneumoniae / PSI-Biology / NYSGRC / Structural Genomics / New York Structural Genomics Research Consortium
Function / homologyNADP-dependent oxidoreductase domain / TIM Barrel / Alpha-Beta Barrel / Alpha Beta / ACETATE ION / NADP NICOTINAMIDE-ADENINE-DINUCLEOTIDE PHOSPHATE / :
Function and homology information
Biological speciesKlebsiella pneumoniae (bacteria)
MethodX-RAY DIFFRACTION / SYNCHROTRON / SAD / Resolution: 1.8 Å
AuthorsBacal, P. / Shabalin, I.G. / Cooper, D.R. / Hillerich, B.S. / Zimmerman, M.D. / Chowdhury, S. / Hammonds, J. / Al Obaidi, N. / Gizzi, A. / Bonanno, J. ...Bacal, P. / Shabalin, I.G. / Cooper, D.R. / Hillerich, B.S. / Zimmerman, M.D. / Chowdhury, S. / Hammonds, J. / Al Obaidi, N. / Gizzi, A. / Bonanno, J. / Seidel, R. / Almo, S.C. / Minor, W. / New York Structural Genomics Research Consortium (NYSGRC)
Funding support United States, 1items
OrganizationGrant numberCountry
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)U54-GM094662 United States
CitationJournal: to be published
Title: Crystal structure of aldo/keto reductase from Klebsiella pneumoniae in complex with NADP and acetate at 1.8 A resolution
Authors: Bacal, P. / Shabalin, I.G. / Cooper, D.R. / Hillerich, B.S. / Zimmerman, M.D. / Bonanno, J. / Seidel, R. / Almo, S.C. / Minor, W.
History
DepositionSep 18, 2014Deposition site: RCSB / Processing site: RCSB
Revision 1.0Oct 1, 2014Provider: repository / Type: Initial release
Revision 1.1Oct 14, 2015Group: Data collection
Revision 1.2Nov 22, 2017Group: Derived calculations / Refinement description / Category: pdbx_struct_oper_list / software / Item: _pdbx_struct_oper_list.symmetry_operation
Revision 1.3Nov 27, 2019Group: Author supporting evidence / Category: pdbx_audit_support
Item: _pdbx_audit_support.funding_organization / _pdbx_audit_support.grant_number
Revision 1.4Apr 13, 2022Group: Database references / Structure summary / Category: audit_author / citation_author / database_2
Item: _audit_author.identifier_ORCID / _citation_author.identifier_ORCID ..._audit_author.identifier_ORCID / _citation_author.identifier_ORCID / _database_2.pdbx_DOI / _database_2.pdbx_database_accession
Revision 1.5Dec 27, 2023Group: Data collection / Category: chem_comp_atom / chem_comp_bond

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

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Assembly

Deposited unit
A: Aldehyde reductase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)35,5713
Polymers34,7691
Non-polymers8022
Water4,053225
1


  • Idetical with deposited unit
  • defined by author
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Unit cell
Length a, b, c (Å)99.483, 99.483, 55.181
Angle α, β, γ (deg.)90.000, 90.000, 120.000
Int Tables number171
Space group name H-MP62
Components on special symmetry positions
IDModelComponents
11A-455-

HOH

Detailsbiological unit is the same as asym.

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Components

#1: Protein Aldehyde reductase


Mass: 34768.977 Da / Num. of mol.: 1 / Mutation: T279N
Source method: isolated from a genetically manipulated source
Details: Mutatation: T279N / Source: (gene. exp.) Klebsiella pneumoniae (bacteria) / Gene: L364_03163, WP_004176538.1 / Plasmid: pSGC-His / Production host: Escherichia coli (E. coli) / Strain (production host): BL21 (DE3) RIL / References: UniProt: V3S9R5
#2: Chemical ChemComp-NAP / NADP NICOTINAMIDE-ADENINE-DINUCLEOTIDE PHOSPHATE / 2'-MONOPHOSPHOADENOSINE 5'-DIPHOSPHORIBOSE


Mass: 743.405 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: C21H28N7O17P3
#3: Chemical ChemComp-ACT / ACETATE ION


Mass: 59.044 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: C2H3O2
#4: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 225 / 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: 2.3 Å3/Da / Density % sol: 46.55 %
Crystal growTemperature: 289 K / Method: vapor diffusion, hanging drop / pH: 6.2
Details: 0.45 ul of 13 mg/ml protein in 20 mM HEPES pH 7.5, 150 mM NaCl, 10% Glycerol, 0.1% Sodium Azide, 0.5 mM TCEP and 30mM NADP were mixed with 0.45 ul of 2.8 M Sodium Acetate pH 6.2 and ...Details: 0.45 ul of 13 mg/ml protein in 20 mM HEPES pH 7.5, 150 mM NaCl, 10% Glycerol, 0.1% Sodium Azide, 0.5 mM TCEP and 30mM NADP were mixed with 0.45 ul of 2.8 M Sodium Acetate pH 6.2 and equilibrated against 1.5 M NaCl solution in 96 Well 3 drop Crystallization Plate (Swissci).

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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: Mar 20, 2014 / 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: 1.8→50 Å / Num. obs: 56317 / % possible obs: 99.1 % / Observed criterion σ(I): -3 / Redundancy: 3.9 % / Biso Wilson estimate: 28.4 Å2 / Rmerge(I) obs: 0.048 / Rpim(I) all: 0.028 / Rrim(I) all: 0.056 / Χ2: 0.978 / Net I/av σ(I): 27.4 / Net I/σ(I): 10.2 / Num. measured all: 218509
Reflection shell

Diffraction-ID: 1 / Rejects: _

Resolution (Å)Redundancy (%)Rmerge(I) obsMean I/σ(I) obsNum. unique allCC1/2Rpim(I) allRrim(I) allΧ2% possible all
1.8-1.833.70.621.4328090.7260.3690.7230.64997.3
1.83-1.863.80.51627860.8030.3070.6040.65998.4
1.86-1.93.90.45727810.8190.270.5340.66498.3
1.9-1.943.90.33727780.90.1990.3940.67799.5
1.94-1.983.90.29328340.9190.1720.3410.68698.5
1.98-2.033.90.22228410.9590.130.2580.69599.4
2.03-2.083.90.18827870.9680.110.2190.70999.3
2.08-2.133.90.14428170.980.0840.1670.70298.9
2.13-2.23.90.11528480.9870.0670.1340.71799.5
2.2-2.273.90.11328030.9840.0660.1310.75499.6
2.27-2.353.90.08827960.9920.0510.1020.79399.2
2.35-2.443.90.07428340.9950.0420.0850.83799.5
2.44-2.553.90.06928220.9950.0390.080.88799.5
2.55-2.693.90.05528450.9970.0310.0630.93799.6
2.69-2.863.90.04728300.9970.0270.0551.0699.7
2.86-3.083.90.04328410.9980.0240.0491.20499.6
3.08-3.393.90.04128050.9970.0230.0471.57799.9
3.39-3.883.90.03628360.9980.020.0411.77899.8
3.88-4.883.90.03228480.9980.0180.0371.75599.9
4.88-503.80.0327760.9980.0180.0351.78997.4

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Processing

Software
NameVersionClassification
Blu-Icedata collection
HKL-3000data collection
HKL-3000data reduction
HKL-3000data scaling
HKL-3000phasing
DMphasing
MLPHAREphasing
REFMAC5.8.0073refinement
PDB_EXTRACT3.15data extraction
DENZOdata reduction
SCALEPACKdata scaling
RefinementMethod to determine structure: SAD / Resolution: 1.8→50 Å / Cor.coef. Fo:Fc: 0.979 / Cor.coef. Fo:Fc free: 0.967 / WRfactor Rfree: 0.1904 / WRfactor Rwork: 0.1449 / FOM work R set: 0.8622 / SU B: 5.303 / SU ML: 0.082 / SU R Cruickshank DPI: 0.0989 / SU Rfree: 0.1016 / Cross valid method: THROUGHOUT / σ(F): 0 / ESU R: 0.099 / ESU R Free: 0.102 / SU Rfree Cruickshank DPI: 0.1016 / Stereochemistry target values: MAXIMUM LIKELIHOOD
Details: U VALUES : WITH TLS ADDED HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS
RfactorNum. reflection% reflectionSelection details
Rfree0.1854 1473 5.1 %RANDOM
Rwork0.1446 27524 --
obs0.1466 27524 99.87 %-
Solvent computationIon probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.2 Å / Solvent model: MASK
Displacement parametersBiso max: 95.99 Å2 / Biso mean: 38.07 Å2 / Biso min: 19.06 Å2
Baniso -1Baniso -2Baniso -3
1--1.17 Å2-0.59 Å2-0 Å2
2---1.17 Å20 Å2
3---3.8 Å2
Refinement stepCycle: final / Resolution: 1.8→50 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms2166 0 52 227 2445
Biso mean--30.24 43.96 -
Num. residues----283
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.0140.0192287
X-RAY DIFFRACTIONr_bond_other_d0.0020.022163
X-RAY DIFFRACTIONr_angle_refined_deg1.6541.9853126
X-RAY DIFFRACTIONr_angle_other_deg0.84634938
X-RAY DIFFRACTIONr_dihedral_angle_1_deg5.8315288
X-RAY DIFFRACTIONr_dihedral_angle_2_deg34.64923.43499
X-RAY DIFFRACTIONr_dihedral_angle_3_deg14.03315333
X-RAY DIFFRACTIONr_dihedral_angle_4_deg17.8871518
X-RAY DIFFRACTIONr_chiral_restr0.0920.2351
X-RAY DIFFRACTIONr_gen_planes_refined0.0080.0212603
X-RAY DIFFRACTIONr_gen_planes_other0.0010.02534
X-RAY DIFFRACTIONr_mcbond_it1.021.5111138
X-RAY DIFFRACTIONr_mcbond_other1.0121.5071136
X-RAY DIFFRACTIONr_mcangle_it1.512.2561421
LS refinement shellResolution: 1.8→1.847 Å / Total num. of bins used: 20
RfactorNum. reflection% reflection
Rfree0.283 96 -
Rwork0.255 2057 -
all-2153 -
obs--99.95 %
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)
12.81340.3088-0.38157.29624.39485.4163-0.0999-0.2267-0.25550.26120.1496-0.80790.54070.3461-0.04980.11770.03990.00570.1420.11310.23971.6548.95711.875
28.52921.01471.41843.3780.79342.084-0.13940.55140.2487-0.67130.0260.45710.0039-0.18010.11340.2256-0.0237-0.07420.16010.05150.085250.5613.304-0.358
31.62820.59120.0093.00930.33980.75780.02820.0629-0.1474-0.1559-0.09060.00520.1967-0.09850.06240.1956-0.0277-0.00610.15320.040.053855.3989.0435.654
43.34923.1016-0.304210.8023-2.61955.0839-0.0793-0.0580.2443-0.16680.00250.50860.1186-0.39850.07680.1149-0.0121-0.01870.17150.01570.076249.12914.7697.754
521.72898.11871.32923.8014-0.79698.7019-0.0191-0.198-0.6513-0.0336-0.1944-0.22770.7853-0.1930.21350.1950.00010.04670.12860.04710.076954.5576.26716.753
60.32080.1493-0.57177.0032-2.11222.06850.0835-0.10280.0522-0.0156-0.10.0508-0.07650.05680.01650.119-0.014-0.00780.16060.01360.102358.24424.8669.965
719.20177.5037-0.605728.65441.46780.13210.2182-0.36430.48411.0531-0.21430.37090.05440.0104-0.00390.302-0.0290.02080.2370.00780.092654.85529.99316.427
80.81980.4684-1.20493.75-1.92383.43340.0204-0.05090.02590.162-0.0599-0.0333-0.0766-0.08090.03950.1192-0.0167-0.01610.14370.01550.054956.36421.76512.92
91.92861.2417-0.60025.881-2.98754.37210.0749-0.30280.25920.5862-0.15-0.1194-0.4230.05890.07510.1826-0.0438-0.06760.1723-0.00380.076363.67831.34317.534
101.3110.0464-0.68352.25860.25332.3954-0.0021-0.15110.0849-0.1419-0.1207-0.706-0.16060.30740.12290.1265-0.0210.02260.19690.10920.279272.28424.4126.264
113.5204-1.04020.92181.30225.373532.7349-0.0358-0.19930.1133-0.04410.1531-0.168-0.10930.6207-0.11720.21390.00270.23430.26060.17830.893180.44422.5721.796
124.16270.453-1.07952.1499-0.7431.9715-0.0203-0.240.1161-0.1954-0.2242-0.57460.12880.12340.24450.13440.03170.08010.12580.12040.230471.51317.3972.558
139.931619.2324-1.45538.0645-5.61319.931-1.0180.7180.5472-1.70671.47381.1192-0.6091-0.6223-0.45590.65030.06320.12320.51390.26620.367762.00914.635-11.92
147.0676-7.96522.628712.8989-8.849910.14850.49260.87450.6536-0.4701-0.1277-0.3355-0.5468-0.6642-0.36491.0762-0.50140.07831.06140.15130.41367.9968.241-17.699
157.1873-7.2095-5.143331.6741.29755.7402-0.47370.7581-0.038-1.28680.4731-1.4790.2242-0.11230.00060.5275-0.15050.24880.5105-0.13620.244972.853-0.03-15.824
164.54031.1977-0.10696.7022.60144.6307-0.16120.34080.0627-0.3804-0.0653-0.39870.41870.21390.22650.24010.03360.18890.11360.06880.173272.4644.644-5.614
177.08481.93851.33113.77961.29692.093-0.11890.13410.1308-0.3734-0.071-0.2020.27230.00040.18990.19660.00060.06360.12850.07040.085663.4598.050.361
184.9856-0.1912-0.68721.6855-0.2291.5021-0.24520.1073-0.5135-0.47720.0773-0.61250.33570.06880.16790.29920.02520.20540.1440.01290.317572.9352.325-4.818
1910.58519.4913-3.67112.6941-0.09996.25110.03470.9420.6123-0.04840.2011-0.35920.57060.1047-0.23580.42190.1450.35230.56490.35380.562980.76817.448-13.007
2024.34315.93278.496922.2485-4.415810.37110.28361.28921.1551-0.6667-0.11260.21640.15510.0931-0.1710.2520.09580.19150.21530.19650.25770.35226.318-6.939
Refinement TLS group
IDRefine-IDRefine TLS-IDAuth asym-IDAuth seq-ID
1X-RAY DIFFRACTION1A2 - 17
2X-RAY DIFFRACTION2A18 - 31
3X-RAY DIFFRACTION3A32 - 56
4X-RAY DIFFRACTION4A57 - 67
5X-RAY DIFFRACTION5A68 - 74
6X-RAY DIFFRACTION6A75 - 88
7X-RAY DIFFRACTION7A89 - 95
8X-RAY DIFFRACTION8A96 - 116
9X-RAY DIFFRACTION9A117 - 135
10X-RAY DIFFRACTION10A136 - 176
11X-RAY DIFFRACTION11A177 - 184
12X-RAY DIFFRACTION12A185 - 197
13X-RAY DIFFRACTION13A198 - 203
14X-RAY DIFFRACTION14A204 - 210
15X-RAY DIFFRACTION15A211 - 218
16X-RAY DIFFRACTION16A219 - 234
17X-RAY DIFFRACTION17A235 - 247
18X-RAY DIFFRACTION18A248 - 269
19X-RAY DIFFRACTION19A270 - 278
20X-RAY DIFFRACTION20A279 - 284

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