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- PDB-4ywj: Crystal structure of 4-hydroxy-tetrahydrodipicolinate reductase (... -

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Basic information

Entry
Database: PDB / ID: 4ywj
TitleCrystal structure of 4-hydroxy-tetrahydrodipicolinate reductase (HTPA reductase) from Pseudomonas aeruginosa
Components4-hydroxy-tetrahydrodipicolinate reductase
KeywordsOXIDOREDUCTASE / SSGCID / Pseudomonas aeruginosa / 4-hydroxy-tetrahydrodipicolinate reductase / HTPA reductase / NAD / Structural Genomics / Seattle Structural Genomics Center for Infectious Disease
Function / homology
Function and homology information


4-hydroxy-tetrahydrodipicolinate reductase / oxidoreductase activity, acting on CH or CH2 groups, NAD or NADP as acceptor / 4-hydroxy-tetrahydrodipicolinate reductase / diaminopimelate biosynthetic process / lysine biosynthetic process via diaminopimelate / NAD binding / NADP binding / cytosol
Similarity search - Function
Dihydrodipicolinate reductase, conserved site / Dihydrodipicolinate reductase signature. / Dihydrodipicolinate reductase, C-terminal / Dihydrodipicolinate reductase / Dihydrodipicolinate reductase, C-terminus / Dihydrodipicolinate reductase, N-terminal / Dihydrodipicolinate reductase, N-terminus / Dihydrodipicolinate Reductase; domain 2 / Dihydrodipicolinate Reductase; domain 2 / NAD(P)-binding Rossmann-like Domain ...Dihydrodipicolinate reductase, conserved site / Dihydrodipicolinate reductase signature. / Dihydrodipicolinate reductase, C-terminal / Dihydrodipicolinate reductase / Dihydrodipicolinate reductase, C-terminus / Dihydrodipicolinate reductase, N-terminal / Dihydrodipicolinate reductase, N-terminus / Dihydrodipicolinate Reductase; domain 2 / Dihydrodipicolinate Reductase; domain 2 / NAD(P)-binding Rossmann-like Domain / NAD(P)-binding domain superfamily / Rossmann fold / 2-Layer Sandwich / 3-Layer(aba) Sandwich / Alpha Beta
Similarity search - Domain/homology
1,4-BUTANEDIOL / NICOTINAMIDE-ADENINE-DINUCLEOTIDE / TRIETHYLENE GLYCOL / 4-hydroxy-tetrahydrodipicolinate reductase
Similarity search - Component
Biological speciesPseudomonas aeruginosa (bacteria)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 1.8 Å
AuthorsSeattle Structural Genomics Center for Infectious Disease (SSGCID)
CitationJournal: to be published
Title: Crystal structure of 4-hydroxy-tetrahydrodipicolinate reductase (HTPA reductase) from Pseudomonas aeruginosa
Authors: Abendroth, J. / Lorimer, D.D. / Edwards, T.E.
History
DepositionMar 20, 2015Deposition site: RCSB / Processing site: RCSB
Revision 1.0Apr 1, 2015Provider: repository / Type: Initial release
Revision 1.1Sep 27, 2023Group: Data collection / Database references ...Data collection / Database references / Derived calculations / Refinement description / Source and taxonomy
Category: chem_comp_atom / chem_comp_bond ...chem_comp_atom / chem_comp_bond / database_2 / entity_src_gen / pdbx_initial_refinement_model / pdbx_prerelease_seq / pdbx_struct_oper_list
Item: _database_2.pdbx_DOI / _database_2.pdbx_database_accession ..._database_2.pdbx_DOI / _database_2.pdbx_database_accession / _entity_src_gen.pdbx_alt_source_flag / _pdbx_struct_oper_list.symmetry_operation

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: 4-hydroxy-tetrahydrodipicolinate reductase
B: 4-hydroxy-tetrahydrodipicolinate reductase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)60,6289
Polymers58,7852
Non-polymers1,8437
Water6,413356
1
A: 4-hydroxy-tetrahydrodipicolinate reductase
B: 4-hydroxy-tetrahydrodipicolinate reductase
hetero molecules

A: 4-hydroxy-tetrahydrodipicolinate reductase
B: 4-hydroxy-tetrahydrodipicolinate reductase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)121,25618
Polymers117,5704
Non-polymers3,68614
Water724
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
crystal symmetry operation4_565x,-y+1,-z1
Buried area11940 Å2
ΔGint-9 kcal/mol
Surface area40520 Å2
MethodPISA
Unit cell
Length a, b, c (Å)93.010, 101.500, 144.510
Angle α, β, γ (deg.)90.000, 90.000, 90.000
Int Tables number20
Space group name H-MC2221
Components on special symmetry positions
IDModelComponents
11A-550-

HOH

21A-585-

HOH

31B-538-

HOH

41B-565-

HOH

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Components

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Protein , 1 types, 2 molecules AB

#1: Protein 4-hydroxy-tetrahydrodipicolinate reductase / / HTPA reductase


Mass: 29392.570 Da / Num. of mol.: 2
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Pseudomonas aeruginosa (strain ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228) (bacteria)
Strain: ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228 / Gene: dapB, PA4759 / Plasmid: PsaeA.00820.a.B1 / Production host: Escherichia coli (E. coli) / Strain (production host): BL21(DE3)
References: UniProt: P38103, 4-hydroxy-tetrahydrodipicolinate reductase

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Non-polymers , 5 types, 363 molecules

#2: Chemical ChemComp-NAD / NICOTINAMIDE-ADENINE-DINUCLEOTIDE / Nicotinamide adenine dinucleotide


Mass: 663.425 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: C21H27N7O14P2 / Comment: NAD*YM
#3: Chemical ChemComp-PGE / TRIETHYLENE GLYCOL / Polyethylene glycol


Mass: 150.173 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: C6H14O4
#4: Chemical ChemComp-BU1 / 1,4-BUTANEDIOL / 1,4-Butanediol


Mass: 90.121 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: C4H10O2
#5: Chemical ChemComp-CL / CHLORIDE ION / Chloride


Mass: 35.453 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: Cl
#6: Water ChemComp-HOH / water / Water


Mass: 18.015 Da / Num. of mol.: 356 / 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: 2.91 Å3/Da / Density % sol: 58 %
Crystal growTemperature: 290 K / Method: vapor diffusion, sitting drop / pH: 8.5
Details: Morpheus D9: 20mM of each: 1,6-Hexanediol; 1-Butanol, 1,2-Propanediol (racemic); 2-Propanol; 1,4-Butanediol; 1,3-Propanediol; 100mM Tris Base / Bicine pH 8.5, 30% each PEG 550MME, PEG 20000; ...Details: Morpheus D9: 20mM of each: 1,6-Hexanediol; 1-Butanol, 1,2-Propanediol (racemic); 2-Propanol; 1,4-Butanediol; 1,3-Propanediol; 100mM Tris Base / Bicine pH 8.5, 30% each PEG 550MME, PEG 20000; PsaeA.00820.a.B1.PW37541 at 22.65 mg/ml; cryo: direct; tray 259740d90, puck hxm4-4

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: APS / Beamline: 21-ID-G / Wavelength: 0.97856 Å
DetectorType: RAYONIX MX-300 / Detector: CCD / Date: Feb 25, 2014
RadiationMonochromator: Diamond [111] / Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.97856 Å / Relative weight: 1
ReflectionResolution: 1.8→50 Å / Num. obs: 63326 / % possible obs: 99.4 % / Observed criterion σ(I): -3 / Redundancy: 5 % / Biso Wilson estimate: 24.1 Å2 / Rmerge F obs: 0.999 / Rmerge(I) obs: 0.048 / Rrim(I) all: 0.053 / Χ2: 1.035 / Net I/σ(I): 18.11 / Num. measured all: 316725
Reflection shell

Diffraction-ID: 1 / Rejects: 0

Resolution (Å)Redundancy (%)Rmerge F obsRmerge(I) obsMean I/σ(I) obsNum. measured obsNum. possibleNum. unique obsRrim(I) all% possible all
1.8-1.855.20.8370.5172.924010465646540.578100
1.85-1.90.9140.4193.5523598456945620.46899.8
1.9-1.950.9490.2985.0422630440844010.33399.8
1.95-2.010.9720.2266.5521785428442780.25299.9
2.01-2.080.9750.1897.9221287418941820.2199.8
2.08-2.150.9820.1589.320353404440400.17699.9
2.15-2.230.9880.12311.6419435388538790.13799.8
2.23-2.320.9910.10113.9518666377137550.11399.6
2.32-2.430.9930.08815.9317719358435720.09899.7
2.43-2.550.9960.07218.7716764343734260.0899.7
2.55-2.680.9960.06420.5616064331032880.07299.3
2.68-2.850.9970.05324.7814907311830860.05999
2.85-3.040.9980.04428.8414015292529000.04999.1
3.04-3.290.9990.03832.8213096275427210.04398.8
3.29-3.60.9990.03238.6512149252024910.03698.8
3.6-4.020.9990.02842.5411258229722730.03199
4.02-4.650.9990.02446.0510234205520300.02698.8
4.65-5.6910.02245.978743174317170.02498.5
5.69-8.0510.0245.746665135913260.02397.6
8.05-500.9990.01846.3433478087450.0292.2

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Processing

Software
NameVersionClassification
XDSdata reduction
PHASERphasing
ARPmodel building
Cootmodel building
PHENIXdev_1980refinement
PDB_EXTRACTdata extraction
XSCALEdata scaling
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: 1ARZ
Resolution: 1.8→34.287 Å / SU ML: 0.22 / Cross valid method: FREE R-VALUE / σ(F): 1.34 / Phase error: 21.21 / Stereochemistry target values: ML
RfactorNum. reflection% reflectionSelection details
Rfree0.2006 3153 5.01 %Random selection
Rwork0.1677 59774 --
obs0.1694 62927 99.1 %-
Solvent computationShrinkage radii: 0.9 Å / VDW probe radii: 1.11 Å / Solvent model: FLAT BULK SOLVENT MODEL
Displacement parametersBiso max: 108.18 Å2 / Biso mean: 36.7854 Å2 / Biso min: 10.95 Å2
Refinement stepCycle: final / Resolution: 1.8→34.287 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms3872 0 121 356 4349
Biso mean--40.04 39.06 -
Num. residues----536
Refine LS restraints
Refine-IDTypeDev idealNumber
X-RAY DIFFRACTIONf_bond_d0.0074111
X-RAY DIFFRACTIONf_angle_d0.9915580
X-RAY DIFFRACTIONf_chiral_restr0.048655
X-RAY DIFFRACTIONf_plane_restr0.004729
X-RAY DIFFRACTIONf_dihedral_angle_d15.8961466
LS refinement shell
Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkRefine-ID% reflection obs (%)
1.8-1.82690.32281260.27812425X-RAY DIFFRACTION94
1.8269-1.85540.27731550.26392586X-RAY DIFFRACTION100
1.8554-1.88580.28061310.24352571X-RAY DIFFRACTION100
1.8858-1.91830.28041340.21742619X-RAY DIFFRACTION100
1.9183-1.95320.24681290.2042591X-RAY DIFFRACTION100
1.9532-1.99080.2191280.19722566X-RAY DIFFRACTION100
1.9908-2.03140.21381580.18682614X-RAY DIFFRACTION100
2.0314-2.07560.25181380.19132566X-RAY DIFFRACTION100
2.0756-2.12390.19451300.192609X-RAY DIFFRACTION100
2.1239-2.1770.21400.18762578X-RAY DIFFRACTION100
2.177-2.23580.23181390.18752599X-RAY DIFFRACTION100
2.2358-2.30160.26121420.17932610X-RAY DIFFRACTION100
2.3016-2.37590.20731340.17882615X-RAY DIFFRACTION100
2.3759-2.46080.21741460.16552602X-RAY DIFFRACTION100
2.4608-2.55920.19751290.16332617X-RAY DIFFRACTION100
2.5592-2.67570.21281330.17452589X-RAY DIFFRACTION99
2.6757-2.81670.20751470.17342581X-RAY DIFFRACTION99
2.8167-2.99310.22211330.17282631X-RAY DIFFRACTION99
2.9931-3.2240.19711360.17322582X-RAY DIFFRACTION99
3.224-3.54810.18321400.1592625X-RAY DIFFRACTION99
3.5481-4.06090.16971310.14182653X-RAY DIFFRACTION99
4.0609-5.11350.15331350.12852642X-RAY DIFFRACTION99
5.1135-500.18191390.15392703X-RAY DIFFRACTION97
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)
14.25040.0212-0.0553.9624-0.91545.5622-0.0037-0.19890.66630.25770.0037-0.4499-0.12310.7910.0150.20260.0561-0.0230.3963-0.04340.429648.038343.725514.5955
21.18990.2519-0.27163.10960.10423.4887-0.0791-0.09530.759-0.0446-0.0628-0.6955-0.17031.04760.04440.28240.03030.01340.6359-0.06240.683855.106444.270913.2717
32.64220.38291.6660.7444-1.03683.3974-0.0443-0.16570.38020.2123-0.0174-0.8687-0.06061.10570.02130.30270.1211-0.04960.6293-0.06560.55753.904240.005116.5465
41.7298-0.54350.60721.40210.19291.7454-0.1043-0.4092-0.22460.31760.1943-0.41890.62930.4734-0.06230.44680.367-0.06690.4351-0.04920.363348.366228.336617.5002
51.1412-0.44810.20870.53570.10311.157-0.1681-0.1624-0.27540.15040.1454-0.11840.37070.1339-0.0520.28330.07550.02880.13910.00890.245728.319733.005110.1336
61.71120.7256-0.22411.1762-0.3251.1671-0.0614-0.0826-0.6630.1177-0.0129-0.10580.54520.0209-0.03380.34110.02210.05710.13550.0190.349618.879428.02076.5204
71.3094-0.08140.03590.7218-0.09731.4218-0.1170.0518-0.285-0.07260.0804-0.04760.2303-0.11740.04170.1993-0.01870.02840.1177-0.01850.181916.285438.1943-0.448
82.7259-0.68530.16653.1008-1.31993.472-0.1643-0.470.07440.22740.19290.02290.34330.23620.00350.26670.1388-0.00620.2789-0.04110.208736.290537.64720.3712
93.388-0.04821.45013.257-0.06042.8184-0.04650.4393-0.1776-0.1830.33651.0640.202-0.7189-0.12860.26120.0025-0.00930.73560.09290.5413-15.283944.577312.7038
103.56381.06352.88391.30270.89082.47660.102-0.04120.00460.09460.06481.07190.1805-0.9528-0.23870.37340.01010.10410.73540.07210.66-18.011744.676317.6485
111.0789-0.01110.14851.4066-0.10981.4967-0.143-0.36050.11020.30710.1170.0825-0.0804-0.2463-0.04720.22570.09260.02390.2486-0.04480.12859.86154.394818.9743
122.03230.0706-0.61372.377-0.78715.4377-0.2027-0.4054-0.2130.37280.16920.16490.2808-0.1670.07330.27010.02390.07960.40060.04870.2319-0.349541.551721.6489
Refinement TLS group
IDRefine-IDRefine TLS-IDSelection detailsAuth asym-IDAuth seq-ID
1X-RAY DIFFRACTION1chain 'A' and (resid 1 through 22 )A0
2X-RAY DIFFRACTION2chain 'A' and (resid 23 through 49 )A0
3X-RAY DIFFRACTION3chain 'A' and (resid 50 through 78 )A0
4X-RAY DIFFRACTION4chain 'A' and (resid 79 through 104 )A0
5X-RAY DIFFRACTION5chain 'A' and (resid 105 through 165 )A0
6X-RAY DIFFRACTION6chain 'A' and (resid 166 through 186 )A0
7X-RAY DIFFRACTION7chain 'A' and (resid 187 through 236 )A0
8X-RAY DIFFRACTION8chain 'A' and (resid 237 through 268 )A0
9X-RAY DIFFRACTION9chain 'B' and (resid 1 through 50 )B0
10X-RAY DIFFRACTION10chain 'B' and (resid 51 through 78 )B0
11X-RAY DIFFRACTION11chain 'B' and (resid 79 through 236 )B0
12X-RAY DIFFRACTION12chain 'B' and (resid 237 through 268 )B0

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