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- PDB-2esp: Human ubiquitin-conjugating enzyme (E2) UbcH5b mutant Ile88Ala -

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Basic information

Entry
Database: PDB / ID: 2esp
TitleHuman ubiquitin-conjugating enzyme (E2) UbcH5b mutant Ile88Ala
ComponentsUbiquitin-conjugating enzyme E2 D2
KeywordsLIGASE
Function / homology
Function and homology information


(E3-independent) E2 ubiquitin-conjugating enzyme / E2 ubiquitin-conjugating enzyme / ubiquitin conjugating enzyme activity / protein K48-linked ubiquitination / protein autoubiquitination / ubiquitin ligase complex / Synthesis of active ubiquitin: roles of E1 and E2 enzymes / TICAM1, RIP1-mediated IKK complex recruitment / IKK complex recruitment mediated by RIP1 / Negative regulators of DDX58/IFIH1 signaling ...(E3-independent) E2 ubiquitin-conjugating enzyme / E2 ubiquitin-conjugating enzyme / ubiquitin conjugating enzyme activity / protein K48-linked ubiquitination / protein autoubiquitination / ubiquitin ligase complex / Synthesis of active ubiquitin: roles of E1 and E2 enzymes / TICAM1, RIP1-mediated IKK complex recruitment / IKK complex recruitment mediated by RIP1 / Negative regulators of DDX58/IFIH1 signaling / Peroxisomal protein import / Regulation of TNFR1 signaling / protein modification process / Oxygen-dependent proline hydroxylation of Hypoxia-inducible Factor Alpha / Inactivation of CSF3 (G-CSF) signaling / CLEC7A (Dectin-1) signaling / FCERI mediated NF-kB activation / protein polyubiquitination / ubiquitin-protein transferase activity / ubiquitin protein ligase activity / Antigen processing: Ubiquitination & Proteasome degradation / Downstream TCR signaling / E3 ubiquitin ligases ubiquitinate target proteins / Neddylation / ubiquitin-dependent protein catabolic process / protein ubiquitination / ubiquitin protein ligase binding / protein-containing complex / extracellular exosome / nucleoplasm / ATP binding / nucleus / cytosol
Similarity search - Function
Ubiquitin Conjugating Enzyme / Ubiquitin Conjugating Enzyme / Ubiquitin-conjugating enzyme, active site / Ubiquitin-conjugating (UBC) active site signature. / Ubiquitin-conjugating enzyme E2, catalytic domain homologues / Ubiquitin-conjugating enzyme E2 / Ubiquitin-conjugating enzyme / Ubiquitin-conjugating (UBC) core domain profile. / Ubiquitin-conjugating enzyme/RWD-like / Roll / Alpha Beta
Similarity search - Domain/homology
Ubiquitin-conjugating enzyme E2 D2
Similarity search - Component
Biological speciesHomo sapiens (human)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 1.52 Å
AuthorsOzkan, E. / Yu, H. / Deisenhofer, J.
CitationJournal: Proc.Natl.Acad.Sci.Usa / Year: 2005
Title: Mechanistic insight into the allosteric activation of a ubiquitin-conjugating enzyme by RING-type ubiquitin ligases
Authors: Ozkan, E. / Yu, H. / Deisenhofer, J.
History
DepositionOct 26, 2005Deposition site: RCSB / Processing site: RCSB
Revision 1.0Dec 6, 2005Provider: repository / Type: Initial release
Revision 1.1May 1, 2008Group: Version format compliance
Revision 1.2Jul 13, 2011Group: Advisory / Version format compliance
Revision 1.3Oct 18, 2017Group: Refinement description / Category: software
Revision 1.4Oct 20, 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 1.5Aug 23, 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: Ubiquitin-conjugating enzyme E2 D2
hetero molecules


Theoretical massNumber of molelcules
Total (without water)16,9953
Polymers16,8411
Non-polymers1542
Water2,450136
1


  • Idetical with deposited unit
  • defined by author
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Unit cell
Length a, b, c (Å)47.877, 49.224, 62.128
Angle α, β, γ (deg.)90.00, 90.00, 90.00
Int Tables number19
Space group name H-MP212121

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Components

#1: Protein Ubiquitin-conjugating enzyme E2 D2 / Ubiquitin-protein ligase D2 / Ubiquitin carrier protein D2 / Ubiquitin-conjugating enzyme E2-17 kDa ...Ubiquitin-protein ligase D2 / Ubiquitin carrier protein D2 / Ubiquitin-conjugating enzyme E2-17 kDa 2 / E217 / KB 2


Mass: 16841.277 Da / Num. of mol.: 1 / Mutation: I88A
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Homo sapiens (human) / Gene: UBE2D2, UBC4, UBCH5B / Plasmid: pHis-parallel / Species (production host): Escherichia coli / Production host: Escherichia coli BL21(DE3) (bacteria) / Strain (production host): BL21(DE3) / References: UniProt: P62837, ubiquitin-protein ligase
#2: Chemical ChemComp-CL / CHLORIDE ION


Mass: 35.453 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: Cl
#3: Chemical ChemComp-MPD / (4S)-2-METHYL-2,4-PENTANEDIOL


Mass: 118.174 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: C6H14O2 / Comment: precipitant*YM
#4: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 136 / 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: 2.2 Å3/Da / Density % sol: 43 %
Crystal growTemperature: 294 K / Method: vapor diffusion, hanging drop / pH: 4.5
Details: NaCl, PEG 6000, Sodium Acetate, temperature 294K, pH 4.5, VAPOR DIFFUSION, HANGING DROP

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: APS / Beamline: 19-BM / Wavelength: 0.97897 Å
DetectorType: SBC-3 / Detector: CCD / Date: Jul 15, 2005
RadiationProtocol: SINGLE WAVELENGTH / Scattering type: x-ray
Radiation wavelengthWavelength: 0.97897 Å / Relative weight: 1
ReflectionResolution: 1.52→50 Å / Num. obs: 22996 / % possible obs: 98.7 % / Redundancy: 4.9 % / Rmerge(I) obs: 0.045 / Net I/σ(I): 31.6
Reflection shellResolution: 1.52→1.57 Å / Redundancy: 3.6 % / Rmerge(I) obs: 0.522 / Mean I/σ(I) obs: 2.2 / % possible all: 91

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Phasing

Phasing MR
Highest resolutionLowest resolution
Rotation2.5 Å34.32 Å
Translation2.5 Å34.32 Å

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Processing

Software
NameVersionClassificationNB
DENZOdata reduction
SCALEPACKdata scaling
PHASERphasing
REFMACrefmac_5.2.0005refinement
PDB_EXTRACT1.7data extraction
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: PDB ENTRY 2ESK
Resolution: 1.52→38.58 Å / Cor.coef. Fo:Fc: 0.964 / Cor.coef. Fo:Fc free: 0.946 / SU B: 2.95 / SU ML: 0.056 / TLS residual ADP flag: LIKELY RESIDUAL / Cross valid method: THROUGHOUT / ESU R: 0.082 / ESU R Free: 0.091 / Stereochemistry target values: MAXIMUM LIKELIHOOD / Details: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS
RfactorNum. reflection% reflectionSelection details
Rfree0.23342 1168 5.1 %RANDOM
Rwork0.18415 ---
obs0.18663 21765 98.76 %-
Solvent computationIon probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.2 Å / Solvent model: BABINET MODEL WITH MASK
Displacement parametersBiso mean: 18.722 Å2
Baniso -1Baniso -2Baniso -3
1--0.18 Å20 Å20 Å2
2---0.35 Å20 Å2
3---0.53 Å2
Refinement stepCycle: LAST / Resolution: 1.52→38.58 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms1184 0 9 136 1329
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.0210.0221261
X-RAY DIFFRACTIONr_bond_other_d
X-RAY DIFFRACTIONr_angle_refined_deg2.0841.9681731
X-RAY DIFFRACTIONr_angle_other_deg
X-RAY DIFFRACTIONr_dihedral_angle_1_deg13.4885160
X-RAY DIFFRACTIONr_dihedral_angle_2_deg32.03623.45555
X-RAY DIFFRACTIONr_dihedral_angle_3_deg13.10115201
X-RAY DIFFRACTIONr_dihedral_angle_4_deg17.492158
X-RAY DIFFRACTIONr_chiral_restr0.1380.2188
X-RAY DIFFRACTIONr_gen_planes_refined0.0120.02982
X-RAY DIFFRACTIONr_gen_planes_other
X-RAY DIFFRACTIONr_nbd_refined0.2120.2606
X-RAY DIFFRACTIONr_nbd_other
X-RAY DIFFRACTIONr_nbtor_refined0.3260.2887
X-RAY DIFFRACTIONr_nbtor_other
X-RAY DIFFRACTIONr_xyhbond_nbd_refined0.1590.2102
X-RAY DIFFRACTIONr_xyhbond_nbd_other
X-RAY DIFFRACTIONr_metal_ion_refined
X-RAY DIFFRACTIONr_metal_ion_other
X-RAY DIFFRACTIONr_symmetry_vdw_refined0.1690.252
X-RAY DIFFRACTIONr_symmetry_vdw_other
X-RAY DIFFRACTIONr_symmetry_hbond_refined0.1390.215
X-RAY DIFFRACTIONr_symmetry_hbond_other
X-RAY DIFFRACTIONr_symmetry_metal_ion_refined
X-RAY DIFFRACTIONr_symmetry_metal_ion_other
X-RAY DIFFRACTIONr_mcbond_it1.1821.5795
X-RAY DIFFRACTIONr_mcbond_other
X-RAY DIFFRACTIONr_mcangle_it1.59521270
X-RAY DIFFRACTIONr_scbond_it2.6623533
X-RAY DIFFRACTIONr_scangle_it4.0354.5456
X-RAY DIFFRACTIONr_rigid_bond_restr
X-RAY DIFFRACTIONr_sphericity_free
X-RAY DIFFRACTIONr_sphericity_bonded
LS refinement shell

Refine-ID: X-RAY DIFFRACTION / Total num. of bins used: 20

Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkNum. reflection all% reflection obs (%)
1.52-1.5590.318860.2721405166889.388
1.559-1.6020.302800.2441521165496.796
1.602-1.6480.302590.2321525159799.186
1.648-1.6990.298780.2231463154699.677
1.699-1.7550.25760.21814421518100
1.755-1.8160.248730.20613911464100
1.816-1.8850.257600.1851364142599.93
1.885-1.9610.217620.1841288135299.852
1.961-2.0480.255610.1691249131699.544
2.048-2.1480.204740.16811921266100
2.148-2.2640.211620.1661124118899.832
2.264-2.4010.236550.1811069113099.469
2.401-2.5660.206630.17710121075100
2.566-2.770.2580.187939997100
2.77-3.0330.218590.19987093299.678
3.033-3.3880.28510.17278984599.408
3.388-3.9080.194320.15172475999.605
3.908-4.7740.225330.17616649100
4.774-6.70.289240.19849351899.807
6.7-38.5760.22220.19328932296.584
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)
114.12834.97422.13344.60311.68534.33760.08630.05670.0605-0.01170.0214-0.32170.06460.231-0.1078-0.04920.02260.0017-0.0149-0.00570.001837.754-6.89758.813
26.86620.67881.45216.8554-1.43151.7220.13230.0270.1769-0.1835-0.0441-0.04160.08240.0461-0.0882-0.03940.00650.0067-0.0337-0.0094-0.033331.441-4.92754.973
315.3181-4.86434.961314.0194-0.00185.78780.13310.5333-0.5185-0.355-0.19150.69910.01050.20980.05840.02040.0149-0.03560.0027-0.03310.029924.296-6.9549.567
443.36129.6947-9.11316.41812.39426.9745-0.94031.4006-0.5279-0.47130.31950.57860.3558-0.62930.62080.0104-0.0615-0.01060.1122-0.0326-0.066516.116-2.81954.642
55.71575.87081.129911.0855-2.55457.7389-0.16460.45690.3079-0.03640.195-0.4122-0.43680.012-0.03040.00060.00620.0252-0.0276-0.00950.040930.6346.44557.203
612.6755-2.7292.11042.9524-0.96492.2285-0.0236-0.15410.22140.10170.0619-0.37250.09640.0534-0.0383-0.0499-0.0024-0.0011-0.0311-0.0148-0.017630.9532.20160.591
733.000212.37060.79865.89730.95060.8715-0.33910.71530.0539-0.18260.3170.05720.00390.08870.0221-0.0403-0.0045-0.0233-0.01710.0124-0.038510.536-0.67159.193
84.94090.98790.73035.4115-0.73651.18910.05230.29640.2514-0.05820.06650.1987-0.00610.1245-0.1188-0.04760.0124-0.0098-0.0187-0.0026-0.04468.1862.75461.921
93.3101-2.0873-1.03033.36820.76050.3267-0.0144-0.00410.0890.06740.0309-0.13720.03850.0209-0.0164-0.04270.0005-0.0242-0.0033-0.0095-0.027130.479-2.2865.352
1033.8551-30.59671.638630.6176-0.63811.5964-0.2872-0.04060.22180.54370.203-0.17090.11290.00520.0842-0.01790.01550.00170.00770.0101-0.058729.204-5.26668.881
1114.9531-1.3578-4.56413.7635-3.54765.7055-0.2206-0.44910.62590.11870.2949-0.2016-0.17380.2765-0.07430.04460.0183-0.05650.047-0.09110.067913.3675.75368.589
122.2306-0.3699-1.9654.94050.07384.8590.0164-0.0984-0.11360.07230.0089-0.013-0.15260.0402-0.0253-0.0060.01540.0076-0.0075-0.0066-0.07449.958-0.58573.871
139.9836-10.6437-5.412822.57944.77824.9421-0.3036-0.134-0.06790.28190.0028-0.22940.09750.06660.30070.0134-0.0027-0.03450.0824-0.0007-0.0418.884-3.16173.175
148.8716-4.306-1.241513.6955.710215.0843-0.1986-0.7768-0.18791.110.15630.25740.1543-0.15950.04230.05840.00150.03580.03470.0131-0.024626.285-12.65769.364
1510.8357-0.3038-2.93810.860.3351.763-0.06580.02540.01030.06710.0418-0.04920.01-0.07060.024-0.038-0.0009-0.0007-0.0517-0.0064-0.055617.41-7.69662.898
165.63183.52890.817610.3778-1.591230.58080.3062-0.4671-0.03991.2751-0.56820.13880.67090.9110.2620.1557-0.02960.0430.0504-0.01910.0129.782-9.5574.903
174.68011.81171.36132.91561.53544.4848-0.0078-0.0978-0.17550.269-0.10570.07120.29840.02730.11340.0235-0.00260.0487-0.0038-0.0206-0.04132.878-3.80378.529
189.5535-3.26543.2636.0041-5.175611.44460.0248-0.1440.12920.1548-0.00310.3352-0.1544-0.4114-0.02170.01150.00540.0578-0.0178-0.05170.0472-2.847-0.50171.869
192.5607-0.4559-0.014812.96113.974713.77990.0615-0.1673-0.01710.6173-0.1860.4587-0.0909-0.47730.1245-0.03450.02430.0261-0.02-0.01370.02790.9995.10969.601
202.45442.1422-1.3839.99362.07895.48660.06050.05160.2111-0.0030.03420.2439-0.1385-0.233-0.0947-0.02360.0233-0.016-0.02490.01190.01655.2189.52864.729
Refinement TLS group
IDRefine-IDRefine TLS-IDAuth asym-IDLabel asym-IDAuth seq-IDLabel seq-ID
1X-RAY DIFFRACTION1AA-1 - 51 - 7
2X-RAY DIFFRACTION2AA6 - 108 - 12
3X-RAY DIFFRACTION3AA11 - 1713 - 19
4X-RAY DIFFRACTION4AA18 - 2320 - 25
5X-RAY DIFFRACTION5AA24 - 2926 - 31
6X-RAY DIFFRACTION6AA30 - 3332 - 35
7X-RAY DIFFRACTION7AA34 - 4336 - 45
8X-RAY DIFFRACTION8AA44 - 5146 - 53
9X-RAY DIFFRACTION9AA52 - 6154 - 63
10X-RAY DIFFRACTION10AA62 - 6864 - 70
11X-RAY DIFFRACTION11AA69 - 7371 - 75
12X-RAY DIFFRACTION12AA74 - 7976 - 81
13X-RAY DIFFRACTION13AA80 - 8982 - 91
14X-RAY DIFFRACTION14AA90 - 9792 - 99
15X-RAY DIFFRACTION15AA98 - 110100 - 112
16X-RAY DIFFRACTION16AA111 - 118113 - 120
17X-RAY DIFFRACTION17AA119 - 131121 - 133
18X-RAY DIFFRACTION18AA132 - 136134 - 138
19X-RAY DIFFRACTION19AA137 - 141139 - 143
20X-RAY DIFFRACTION20AA142 - 147144 - 149

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