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- PDB-3kbo: 2.14 Angstrom Crystal Structure of Putative Oxidoreductase (ycdW)... -

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Entry
Database: PDB / ID: 3kbo
Title2.14 Angstrom Crystal Structure of Putative Oxidoreductase (ycdW) from Salmonella typhimurium in Complex with NADP
ComponentsGlyoxylate/hydroxypyruvate reductase A
KeywordsOXIDOREDUCTASE / NADP / NAD / Structural Genomics / Center for Structural Genomics of Infectious Diseases / CSGID
Function / homology
Function and homology information


hydroxypyruvate reductase (NADH) activity / hydroxypyruvate reductase / glyoxylate reductase (NADP+) / hydroxypyruvate reductase [NAD(P)H] activity / glyoxylate reductase (NADPH) activity / NAD binding / cytosol
Similarity search - Function
Glyoxylate/hydroxypyruvate reductase A, Enterobacterales / D-isomer specific 2-hydroxyacid dehydrogenase, NAD-binding domain / D-isomer specific 2-hydroxyacid dehydrogenase, NAD binding domain / NAD(P)-binding Rossmann-like Domain / NAD(P)-binding domain superfamily / Rossmann fold / 3-Layer(aba) Sandwich / Alpha Beta
Similarity search - Domain/homology
Chem-NDB / Chem-NDP / Glyoxylate/hydroxypyruvate reductase A
Similarity search - Component
Biological speciesSalmonella enterica subsp. enterica serovar Typhimurium (bacteria)
MethodX-RAY DIFFRACTION / SYNCHROTRON / SAD / Resolution: 2.14 Å
AuthorsMinasov, G. / Wawrzak, Z. / Skarina, T. / Onopriyenko, O. / Papazisi, L. / Savchenko, A. / Anderson, W.F. / Center for Structural Genomics of Infectious Diseases (CSGID)
CitationJournal: To be Published
Title: 2.14 Angstrom Crystal Structure of Putative Oxidoreductase (ycdW) from Salmonella typhimurium in Complex with NADP.
Authors: Minasov, G. / Wawrzak, Z. / Skarina, T. / Onopriyenko, O. / Papazisi, L. / Savchenko, A. / Anderson, W.F. / Center for Structural Genomics of Infectious Diseases (CSGID)
History
DepositionOct 20, 2009Deposition site: RCSB / Processing site: RCSB
Revision 1.0Oct 27, 2009Provider: repository / Type: Initial release
Revision 1.1Jul 13, 2011Group: Advisory / Refinement description / Version format compliance
Revision 1.2Nov 1, 2017Group: Refinement description / Category: software / Item: _software.name

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: Glyoxylate/hydroxypyruvate reductase A
B: Glyoxylate/hydroxypyruvate reductase A
C: Glyoxylate/hydroxypyruvate reductase A
D: Glyoxylate/hydroxypyruvate reductase A
hetero molecules


Theoretical massNumber of molelcules
Total (without water)146,83719
Polymers142,8824
Non-polymers3,95415
Water9,998555
1
A: Glyoxylate/hydroxypyruvate reductase A
D: Glyoxylate/hydroxypyruvate reductase A
hetero molecules


Theoretical massNumber of molelcules
Total (without water)73,52410
Polymers71,4412
Non-polymers2,0838
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area3780 Å2
ΔGint-20 kcal/mol
Surface area27620 Å2
MethodPISA
2
B: Glyoxylate/hydroxypyruvate reductase A
C: Glyoxylate/hydroxypyruvate reductase A
hetero molecules


Theoretical massNumber of molelcules
Total (without water)73,3129
Polymers71,4412
Non-polymers1,8717
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area3770 Å2
ΔGint-19 kcal/mol
Surface area27580 Å2
MethodPISA
Unit cell
Length a, b, c (Å)279.621, 45.585, 112.335
Angle α, β, γ (deg.)90.00, 101.58, 90.00
Int Tables number5
Space group name H-MC121

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Components

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Protein , 1 types, 4 molecules ABCD

#1: Protein
Glyoxylate/hydroxypyruvate reductase A / 2-ketoacid reductase


Mass: 35720.578 Da / Num. of mol.: 4
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Salmonella enterica subsp. enterica serovar Typhimurium (bacteria)
Strain: LT2 / Gene: ghrA, STM1135, ycdW / Plasmid: pMCSG7 / Production host: Escherichia coli (E. coli) / Strain (production host): BL21-DE3
References: UniProt: Q8ZQ30, glyoxylate reductase (NADP+), hydroxypyruvate reductase

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Non-polymers , 5 types, 570 molecules

#2: Chemical
ChemComp-NDP / NADPH DIHYDRO-NICOTINAMIDE-ADENINE-DINUCLEOTIDE PHOSPHATE


Mass: 745.421 Da / Num. of mol.: 4 / Source method: obtained synthetically / Formula: C21H30N7O17P3
#3: Chemical ChemComp-NDB / N-(2-hydroxyethyl)-N,N-dimethyl-3-sulfopropan-1-aminium


Mass: 212.287 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: C7H18NO4S
#4: Chemical
ChemComp-CL / CHLORIDE ION


Mass: 35.453 Da / Num. of mol.: 8 / Source method: obtained synthetically / Formula: Cl
#5: Chemical ChemComp-EPE / 4-(2-HYDROXYETHYL)-1-PIPERAZINE ETHANESULFONIC ACID / HEPES


Mass: 238.305 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: C8H18N2O4S / Comment: pH buffer*YM
#6: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 555 / 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: 2.45 Å3/Da / Density % sol: 49.88 %
Crystal growTemperature: 295 K / Method: vapor diffusion, hanging drop / pH: 6.5
Details: Protein solution: 0.3M Sodium chloride, 10mM HEPES (pH 7.5); Screen solution: 0.3M NDSB-211, 30% PEG 3350, 0.2M Sodium acetate, 0.1M Bis-Tris pH 6.5, 10mm NADP, VAPOR DIFFUSION, HANGING ...Details: Protein solution: 0.3M Sodium chloride, 10mM HEPES (pH 7.5); Screen solution: 0.3M NDSB-211, 30% PEG 3350, 0.2M Sodium acetate, 0.1M Bis-Tris pH 6.5, 10mm NADP, VAPOR DIFFUSION, HANGING DROP, temperature 295K; Cryo: oil, paratone

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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: Sep 13, 2009 / Details: Beryllium lenses
RadiationMonochromator: Diamond / Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.97872 Å / Relative weight: 1
ReflectionResolution: 2.14→30 Å / Num. all: 75710 / Num. obs: 75710 / % possible obs: 98.2 % / Observed criterion σ(I): -3 / Redundancy: 6.6 % / Biso Wilson estimate: 34.7 Å2 / Rmerge(I) obs: 0.084 / Net I/σ(I): 27
Reflection shellResolution: 2.14→2.18 Å / Redundancy: 6 % / Rmerge(I) obs: 0.491 / Mean I/σ(I) obs: 3.5 / Num. unique all: 3610 / % possible all: 93.5

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Processing

Software
NameVersionClassification
Blu-IceMaxdata collection
PHENIXmodel building
REFMAC5.5.0102refinement
HKL-2000data reduction
HKL-2000data scaling
PHENIXphasing
RefinementMethod to determine structure: SAD / Resolution: 2.14→29.93 Å / Cor.coef. Fo:Fc: 0.957 / Cor.coef. Fo:Fc free: 0.934 / SU B: 9.853 / SU ML: 0.115 / TLS residual ADP flag: LIKELY RESIDUAL / Isotropic thermal model: Individual Isotropic / Cross valid method: THROUGHOUT / ESU R: 0.284 / ESU R Free: 0.229 / Stereochemistry target values: MAXIMUM LIKELIHOOD / Details: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS
RfactorNum. reflection% reflectionSelection details
Rfree0.26879 3806 5 %RANDOM
Rwork0.21092 ---
all0.21387 71864 --
obs0.21387 71864 98 %-
Solvent computationIon probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.4 Å / Solvent model: BABINET MODEL WITH MASK
Displacement parametersBiso mean: 27.209 Å2
Baniso -1Baniso -2Baniso -3
1--1.57 Å20 Å20.23 Å2
2--5.13 Å2-0 Å2
3----3.47 Å2
Refinement stepCycle: LAST / Resolution: 2.14→29.93 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms9900 0 243 555 10698
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.0130.02210493
X-RAY DIFFRACTIONr_bond_other_d0.0010.027108
X-RAY DIFFRACTIONr_angle_refined_deg1.4531.98214344
X-RAY DIFFRACTIONr_angle_other_deg0.87317228
X-RAY DIFFRACTIONr_dihedral_angle_1_deg2.76251262
X-RAY DIFFRACTIONr_dihedral_angle_2_deg30.66423.486479
X-RAY DIFFRACTIONr_dihedral_angle_3_deg11.693151667
X-RAY DIFFRACTIONr_dihedral_angle_4_deg10.6821584
X-RAY DIFFRACTIONr_chiral_restr0.0870.21547
X-RAY DIFFRACTIONr_gen_planes_refined0.0070.02111583
X-RAY DIFFRACTIONr_gen_planes_other0.0010.022149
X-RAY DIFFRACTIONr_mcbond_it1.2861.56290
X-RAY DIFFRACTIONr_mcbond_other0.3751.52514
X-RAY DIFFRACTIONr_mcangle_it2.147210124
X-RAY DIFFRACTIONr_scbond_it3.46934203
X-RAY DIFFRACTIONr_scangle_it4.874.54220
LS refinement shellResolution: 2.14→2.199 Å / Total num. of bins used: 20
RfactorNum. reflection% reflection
Rfree0.323 232 -
Rwork0.297 4985 -
obs-4985 92.53 %
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)
10.46030.11990.00610.3025-0.26630.89410.0407-0.03010.0403-0.0344-0.02270.02520.02680.0321-0.0180.0496-0.00530.0010.00540.00520.052219.3614-0.9043-21.2531
20.43930.12750.26230.08260.16680.355-0.03410.01660.0690.0058-0.00620.01320.0011-0.02370.04040.067-0.0048-0.00810.01260.00570.071422.66375.96039.5326
30.78170.23630.01790.25220.07590.4485-0.05050.02790.10070.02430.00550.0498-0.0099-0.08790.0450.03850.0118-0.01730.02550.00250.068513.24767.194112.9317
40.07190.0390.20870.24750.42281.2802-0.01540.0230.0438-0.01950.01130.00550.0098-0.00990.00410.0433-0.0198-0.02190.03160.04730.075120.03873.6112-4.5158
50.32210.2222-0.29630.93890.18760.82410.0293-0.07030.07310.06450.00420.0274-0.05670.0643-0.03360.0418-0.02090.00470.0581-0.00570.041939.571418.531897.8761
60.15860.03610.00840.0417-0.05180.20510.02050.00640.0094-0.01750.01080.0195-0.01770.036-0.03130.0625-0.0168-0.00130.03070.01320.067128.095515.362666.6234
70.48860.0242-0.01070.173-0.00680.19080.00070.04470.0772-0.0048-0.0065-0.0084-0.0260.05430.00590.0519-0.01370.00260.02130.01040.084837.052122.659965.5416
80.4456-0.12790.23270.1256-0.10940.56720.0171-0.09350.0528-0.01570.01040.0275-0.00230.0799-0.02750.0401-0.00310.00960.0615-0.01850.053437.259113.647584.126
90.65110.0585-0.2760.0419-0.16870.8172-0.0051-0.01110.0008-0.0155-0.00110.0130.00460.00770.00620.0559-0.00320.00130.0004-0.00110.06455.6097-1.95835.8041
100.44770.1580.0460.11750.12780.22020.0079-0.02270.02670.0007-0.01030.00240.0076-0.01690.00230.0590.0005-0.00520.01380.00240.064315.900811.5970.1266
110.41810.0965-0.16450.2208-0.04250.2626-0.02440.05940.02730.02360.00710.0267-0.0181-0.03370.01730.05290.0053-0.00750.01480.01580.06512.8318.663566.3382
120.08820.11880.11720.34440.04660.69070.021-0.0586-0.027-0.0116-0.0715-0.04420.13250.07630.05050.05960.00760.00980.09550.03770.053710.7608-8.980145.0105
130.6030.44710.11240.54870.01490.96220.0533-0.01620.05580.1272-0.00910.0591-0.09810.0294-0.04420.0638-0.00310.03040.02160.00630.025451.314418.658743.0773
140.48380.16940.34630.07270.12160.4524-0.03740.06550.0352-0.02430.03850.0381-0.0650.1146-0.0010.0635-0.0217-0.01330.04960.02560.058940.542714.981311.8914
150.79560.0714-0.03010.3837-0.01210.2979-0.09810.08620.1595-0.05010.06230.0432-0.05960.14220.03580.0531-0.0552-0.01650.0920.03660.061649.54122.154110.4249
160.48320.1440.43290.04350.12960.4066-0.0308-0.00120.0468-0.01230.00170.016-0.02890.03110.02910.03870.00370.01230.0622-0.00330.063848.6814.681928.7217
170.60040.3334-0.8830.2038-0.66893.0135-0.07120.13670.0865-0.02610.06690.035-0.0046-0.10950.00430.0677-0.0053-0.00410.04670.01410.083116.5018-0.632.8276
184.26940.6566-1.86050.1231-0.17181.41170.0844-0.02170.1582-0.0092-0.01070.021-0.1618-0.0076-0.07370.0519-0.01410.00320.05090.00290.063541.024916.709275.4442
190.5569-0.25580.54160.1176-0.24840.5531-0.01350.0522-0.01370.0093-0.02020.0081-0.01360.02190.03370.0762-0.00470.00650.0637-0.03140.0894.459-0.243357.6877
200.1802-0.2529-0.49650.36280.76872.03390.0070.0155-0.0333-0.0288-0.00420.0364-0.16590.1398-0.00280.0801-0.0285-0.00040.0738-0.00830.05753.010416.60120.5402
210.00250.0698-0.351.9495-9.761148.88070.0315-0.0002-0.00030.13640.06720.0271-0.7029-0.1774-0.09870.0929-0.0794-0.01350.13990.01930.13837.1775-3.1945-26.9217
2247.24686.069516.34381.83291.99385.6646-0.35573.57290.43980.02390.23770.3585-0.12851.26550.1180.1261-0.06470.01420.52440.0650.168737.2841.599511.9784
236.65529.757619.064214.318628.564299.55590.26521.0913-0.54630.31171.6264-0.7877-1.73583.1819-1.89160.22770.0233-0.04090.1992-0.03710.186925.48281.970766.6664
Refinement TLS group
IDRefine-IDRefine TLS-IDAuth asym-IDAuth seq-ID
1X-RAY DIFFRACTION1A1 - 67
2X-RAY DIFFRACTION2A68 - 170
3X-RAY DIFFRACTION3A171 - 255
4X-RAY DIFFRACTION4A256 - 312
5X-RAY DIFFRACTION5B1 - 79
6X-RAY DIFFRACTION6B80 - 156
7X-RAY DIFFRACTION7B157 - 265
8X-RAY DIFFRACTION8B266 - 312
9X-RAY DIFFRACTION9C1 - 90
10X-RAY DIFFRACTION10C91 - 156
11X-RAY DIFFRACTION11C157 - 279
12X-RAY DIFFRACTION12C280 - 312
13X-RAY DIFFRACTION13D1 - 79
14X-RAY DIFFRACTION14D80 - 157
15X-RAY DIFFRACTION15D158 - 263
16X-RAY DIFFRACTION16D264 - 312
17X-RAY DIFFRACTION17A313
18X-RAY DIFFRACTION18B313
19X-RAY DIFFRACTION19C313
20X-RAY DIFFRACTION20D313
21X-RAY DIFFRACTION21A314
22X-RAY DIFFRACTION22D314
23X-RAY DIFFRACTION23B314

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