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- PDB-6vdt: Crystal Structure of Dehaloperoxidase B in Complex with cofactor ... -

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Basic information

Entry
Database: PDB / ID: 6vdt
TitleCrystal Structure of Dehaloperoxidase B in Complex with cofactor Iron(III) Deuteroporphyrin IX and Substrate 4-nitrophenol
ComponentsDehaloperoxidase B
KeywordsOXIDOREDUCTASE / heme peroxidase / peroxygenase / heme cofactor / oxygen binding
Function / homology
Function and homology information


oxygen carrier activity / oxygen binding / peroxidase activity / heme binding / metal ion binding
Similarity search - Function
Myoglobin-like, M family globin domain / Globin/Protoglobin / Globin / Globin / Globin-like superfamily
Similarity search - Domain/homology
FE(III) DEUTEROPORPHYRIN IX / P-NITROPHENOL / DI(HYDROXYETHYL)ETHER / Dehaloperoxidase B
Similarity search - Component
Biological speciesAmphitrite ornata (invertebrata)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 1.978 Å
AuthorsGhiladi, R.A. / de Serrano, V.S. / McGuire, A. / Malewschik, T.
Funding support United States, 1items
OrganizationGrant numberCountry
National Science Foundation (NSF, United States) United States
CitationJournal: To Be Published
Title: Nonnative Heme Incorporation into Multifunctional Globin Increases Peroxygenase Activity an Order and Magnitude Compared to Native Enzyme
Authors: McGuire, A.H. / Petit, A.R. / Kang, J. / Malewschik, T. / de Serrano, V. / Carey, L.M. / Ghiladi, R.A.
History
DepositionDec 27, 2019Deposition site: RCSB / Processing site: RCSB
Revision 1.0Dec 30, 2020Provider: repository / Type: Initial release
Revision 1.1Jun 30, 2021Group: Data collection / Category: pdbx_reflns_twin
Item: _pdbx_reflns_twin.crystal_id / _pdbx_reflns_twin.diffrn_id
Revision 1.2Oct 11, 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 / pdbx_validate_chiral
Item: _database_2.pdbx_DOI / _database_2.pdbx_database_accession

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

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Assembly

Deposited unit
A: Dehaloperoxidase B
B: Dehaloperoxidase B
hetero molecules


Theoretical massNumber of molelcules
Total (without water)32,51910
Polymers30,8292
Non-polymers1,6908
Water2,234124
1
A: Dehaloperoxidase B
hetero molecules


Theoretical massNumber of molelcules
Total (without water)16,4787
Polymers15,4141
Non-polymers1,0646
Water181
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
MethodPISA
2
B: Dehaloperoxidase B
hetero molecules


Theoretical massNumber of molelcules
Total (without water)16,0413
Polymers15,4141
Non-polymers6262
Water181
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
MethodPISA
Unit cell
Length a, b, c (Å)58.700, 67.660, 67.730
Angle α, β, γ (deg.)90.000, 90.000, 90.000
Int Tables number19
Space group name H-MP212121

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Components

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Protein , 1 types, 2 molecules AB

#1: Protein Dehaloperoxidase B


Mass: 15414.462 Da / Num. of mol.: 2
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Amphitrite ornata (invertebrata) / Production host: Escherichia coli BL21 (bacteria) / Strain (production host): BL21 / Variant (production host): (DE3) pLysS / References: UniProt: Q9NAV7

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Non-polymers , 6 types, 132 molecules

#2: Chemical ChemComp-FDE / FE(III) DEUTEROPORPHYRIN IX


Mass: 564.413 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: C30H28FeN4O4 / Feature type: SUBJECT OF INVESTIGATION
#3: Chemical ChemComp-EDO / 1,2-ETHANEDIOL / ETHYLENE GLYCOL


Mass: 62.068 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: C2H6O2
#4: Chemical ChemComp-NPO / P-NITROPHENOL


Mass: 139.109 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: C6H5NO3 / Feature type: SUBJECT OF INVESTIGATION
#5: Chemical ChemComp-PEG / DI(HYDROXYETHYL)ETHER


Mass: 106.120 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: C4H10O3
#6: Chemical ChemComp-SO4 / SULFATE ION


Mass: 96.063 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: SO4
#7: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 124 / Source method: isolated from a natural source / Formula: H2O

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Details

Has ligand of interestY

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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: 2.19 Å3/Da / Density % sol: 43.86 %
Crystal growTemperature: 277 K / Method: vapor diffusion, hanging drop / pH: 6.5 / Details: 0.2 M Ammonium Sulphate MPEG 2000, 29-33%

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Data collection

DiffractionMean temperature: 100 K / Serial crystal experiment: N
Diffraction sourceSource: SYNCHROTRON / Site: APS / Beamline: 22-BM / Wavelength: 1 Å
DetectorType: RAYONIX MX300-HS / Detector: CCD / Date: Apr 4, 2019
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 1 Å / Relative weight: 1
Reflection twin
TypeCrystal-IDIDOperatorDomain-IDFraction
pseudo-merohedral11H, K, L10.6286
pseudo-merohedral11-H, -L, -K20.3714
ReflectionResolution: 1.77→47.99 Å / Num. obs: 25924 / % possible obs: 95.57 % / Redundancy: 4.2 % / CC1/2: 0.999 / Net I/σ(I): 20.9
Reflection shellResolution: 1.77→1.82 Å / Num. unique obs: 1325 / CC1/2: 0.6

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Processing

Software
NameVersionClassification
REFMAC5.8.0258refinement
MOSFLMdata reduction
Aimlessdata scaling
PHASERphasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: 3IXF
Resolution: 1.978→37.097 Å / Cor.coef. Fo:Fc: 0.947 / Cor.coef. Fo:Fc free: 0.908 / SU B: 7.579 / SU ML: 0.118 / Cross valid method: FREE R-VALUE / ESU R: 0.05 / ESU R Free: 0.039
Details: Hydrogens have been added in their riding positions
RfactorNum. reflection% reflection
Rfree0.2229 932 5.153 %
Rwork0.167 --
all0.17 --
obs-18087 93.126 %
Solvent computationIon probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.2 Å
Displacement parametersBiso mean: 26.366 Å2
Baniso -1Baniso -2Baniso -3
1-9.816 Å20 Å20 Å2
2---3.008 Å2-0 Å2
3----6.808 Å2
Refinement stepCycle: LAST / Resolution: 1.978→37.097 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms2134 0 113 124 2371
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.0130.0132798
X-RAY DIFFRACTIONr_bond_other_d0.0020.0182458
X-RAY DIFFRACTIONr_angle_refined_deg2.3841.6963857
X-RAY DIFFRACTIONr_angle_other_deg1.4661.6185770
X-RAY DIFFRACTIONr_dihedral_angle_1_deg6.1015378
X-RAY DIFFRACTIONr_dihedral_angle_2_deg32.61422.876153
X-RAY DIFFRACTIONr_dihedral_angle_3_deg19.69915497
X-RAY DIFFRACTIONr_dihedral_angle_4_deg22.291518
X-RAY DIFFRACTIONr_chiral_restr0.140.2339
X-RAY DIFFRACTIONr_gen_planes_refined0.0120.023418
X-RAY DIFFRACTIONr_gen_planes_other0.0070.02646
X-RAY DIFFRACTIONr_nbd_refined0.2750.2825
X-RAY DIFFRACTIONr_symmetry_nbd_other0.2060.22303
X-RAY DIFFRACTIONr_nbtor_refined0.1830.21286
X-RAY DIFFRACTIONr_symmetry_nbtor_other0.0850.21073
X-RAY DIFFRACTIONr_xyhbond_nbd_refined0.2180.2161
X-RAY DIFFRACTIONr_symmetry_xyhbond_nbd_other0.1920.22
X-RAY DIFFRACTIONr_symmetry_nbd_refined0.1530.227
X-RAY DIFFRACTIONr_nbd_other0.2150.293
X-RAY DIFFRACTIONr_symmetry_xyhbond_nbd_refined0.160.217
X-RAY DIFFRACTIONr_mcbond_it1.3691.8421339
X-RAY DIFFRACTIONr_mcbond_other1.3681.841338
X-RAY DIFFRACTIONr_mcangle_it2.2632.761740
X-RAY DIFFRACTIONr_mcangle_other2.2632.7621741
X-RAY DIFFRACTIONr_scbond_it1.6082.2071459
X-RAY DIFFRACTIONr_scbond_other1.5912.1971449
X-RAY DIFFRACTIONr_scangle_it2.4343.3372099
X-RAY DIFFRACTIONr_scangle_other2.4273.3292087
X-RAY DIFFRACTIONr_lrange_it7.37824.5243521
X-RAY DIFFRACTIONr_lrange_other7.3524.4833508
LS refinement shell
Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkRefine-ID% reflection obs (%)
1.978-2.030.346660.2341286X-RAY DIFFRACTION96.3649
2.03-2.0850.335490.18939X-RAY DIFFRACTION70.8752
2.085-2.1450.221550.1661252X-RAY DIFFRACTION99.2407
2.145-2.2110.248530.1671225X-RAY DIFFRACTION100.7092
2.211-2.2830.269550.167838X-RAY DIFFRACTION69.4401
2.283-2.3630.232580.1351170X-RAY DIFFRACTION100.3268
2.363-2.4520.149580.1311117X-RAY DIFFRACTION99.915
2.452-2.5520.196630.1251079X-RAY DIFFRACTION99.5641
2.552-2.6650.172500.1511051X-RAY DIFFRACTION100.182
2.665-2.7940.196410.163816X-RAY DIFFRACTION82.5626
2.794-2.9440.199570.171968X-RAY DIFFRACTION99.9025
2.944-3.1220.289520.17868X-RAY DIFFRACTION100
3.122-3.3350.159520.165842X-RAY DIFFRACTION100
3.335-3.60.208350.179715X-RAY DIFFRACTION87.925
3.6-3.940.188270.16549X-RAY DIFFRACTION75
3.94-4.3990.256350.133676X-RAY DIFFRACTION100.2821
4.399-5.0680.156330.162600X-RAY DIFFRACTION99.8423
5.068-6.1780.231430.22507X-RAY DIFFRACTION99.6377
6.178-8.6180.382300.214407X-RAY DIFFRACTION99.5444
8.618-37.0970.241200.236250X-RAY DIFFRACTION98.5401
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)
10.9811-0.59150.2660.4527-0.39120.6944-0.0270.0115-0.07970.0573-0.010.0515-0.10060.06160.0370.0663-0.00480.00320.08040.01640.0929-0.0264-1.5742-23.8675
25.0535-0.81145.20860.5426-0.71295.4056-0.05120.13970.32130.0629-0.2329-0.2237-0.0370.07870.28410.0312-0.0397-0.02980.13420.07140.141915.7377-5.0262-16.1528
32.85261.5590.47860.85470.26160.0809-0.04750.03160.0598-0.03270.03170.0283-0.01020.00490.01580.0834-0.0142-0.02690.0862-0.03290.066611.3869-1.0885-7.7911
40.576-0.9681-0.01614.02090.20371.5542-0.2894-0.07570.00580.38880.23620.15280.02960.00290.05320.15640.040.01110.06750.02370.06082.8829-5.0058-10.7364
52.14891.457-1.36041.7412-1.72211.71070.3292-0.10590.1610.1443-0.14260.1921-0.12260.1368-0.18660.0903-0.03290.02440.0926-0.02860.0712.33912.904-25.1607
63.5113-4.6858-3.15946.25884.2192.8595-0.2269-0.0414-0.20560.28130.05810.28650.1790.06460.16880.1331-0.0743-0.04510.09640.01980.10758.931313.5743-12.488
73.27893.5908-0.53374.3957-2.15195.39520.2374-0.13780.00750.3548-0.3142-0.0244-0.31160.53470.07680.1119-0.0758-0.08820.08390.06090.106715.23765.7694-18.9093
81.40750.59721.47113.81842.32012.34890.00830.37680.15010.225-0.23180.15360.10440.2090.22350.0524-0.0255-0.00040.1490.04780.09838.3242-3.2946-28.0447
92.03452.83911.2954.65681.15151.63190.01710.0267-0.01560.0309-0.2018-0.16690.06730.1430.18470.0384-0.02960.01010.13570.02660.08417.11313.3734-32.0245
101.0977-0.62532.86162.5723-2.16787.63390.019-0.00220.03250.2012-0.2063-0.57490.0997-0.01470.18740.1895-0.133-0.10490.09590.08180.166914.290715.1292-21.7789
110.56110.3087-0.4420.6237-0.49630.817-0.0502-0.0522-0.0242-0.04610.0582-0.05120.05620.0232-0.0080.0601-0.009-0.01610.07390.0010.0918-17.19796.7463-14.9111
121.7882-0.79183.4556.6677-1.22526.6911-0.0292-0.1404-0.0658-0.12880.12520.5313-0.0573-0.2742-0.0960.0429-0.02320.0160.095-0.06370.1287-27.5398-8.0076-12.9217
136.84671.7605-4.03781.9018-1.48742.6199-0.106-0.1154-0.4052-0.0116-0.2391-0.08940.14130.16080.34520.10050.04960.08750.12060.04550.1478-17.8349-6.9068-5.8743
140.4293-0.6241-1.02334.5209-1.01814.1836-0.0689-0.0175-0.0221-0.11710.12890.16540.2951-0.0448-0.05990.046-0.00280.02540.0760.01620.1044-15.3953-3.597-17.5584
151.5554-1.31090.30923.4114-2.07211.51070.08050.07330.0512-0.1624-0.0821-0.08920.09220.04330.00160.0604-0.01130.00680.0942-0.0080.0788-17.222810.08-29.0125
160.30220.72581.12437.81030.96155.2240.1272-0.013-0.1660.03730.3653-0.17760.0779-0.1625-0.49250.3193-0.0128-0.18890.03260.0040.1779-22.41831.1332-32.9642
172.1652-1.53122.08426.16071.09423.30740.3301-0.1519-0.2007-0.11410.2403-0.57510.3647-0.0762-0.57040.1849-0.07620.00970.0458-0.00490.2201-27.5136-3.9729-26.2835
182.59722.6252.16532.88652.7063.2426-0.03710.06560.20370.0160.1130.14430.0368-0.0152-0.07590.06080.01810.01670.12850.04740.0819-25.07279.713-15.4625
193.72121.63852.82030.99090.81622.81940.0577-0.02940.06750.04320.04410.11660.0144-0.0851-0.10180.0536-0.0058-0.00910.0893-0.01390.0729-22.190815.616-20.3692
203.17951.0767-4.05922.6443-1.17535.2050.08590.2814-0.11460.3376-0.2301-0.0536-0.0432-0.40430.14420.1757-0.1087-0.070.13080.01580.0425-29.91454.8947-31.403
Refinement TLS group
IDRefine-IDRefine TLS-IDSelectionAuth asym-IDAuth seq-ID
1X-RAY DIFFRACTION1ALLAAA1 - 26
2X-RAY DIFFRACTION2ALLAAA27 - 33
3X-RAY DIFFRACTION3ALLAAA34 - 37
4X-RAY DIFFRACTION4ALLAAA38 - 60
5X-RAY DIFFRACTION5ALLAAA61 - 83
6X-RAY DIFFRACTION6ALLAAA84 - 90
7X-RAY DIFFRACTION7ALLAAA91 - 99
8X-RAY DIFFRACTION8ALLAAA100 - 111
9X-RAY DIFFRACTION9ALLAAA112 - 128
10X-RAY DIFFRACTION10ALLAAA129 - 137
11X-RAY DIFFRACTION11ALLBBB1 - 30
12X-RAY DIFFRACTION12ALLBBB31 - 39
13X-RAY DIFFRACTION13ALLBBB40 - 50
14X-RAY DIFFRACTION14ALLBBB51 - 60
15X-RAY DIFFRACTION15ALLBBB61 - 77
16X-RAY DIFFRACTION16ALLBBB78 - 86
17X-RAY DIFFRACTION17ALLBBB87 - 94
18X-RAY DIFFRACTION18ALLBBB95 - 111
19X-RAY DIFFRACTION19ALLBBB112 - 128
20X-RAY DIFFRACTION20ALLBBB129 - 137

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