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- PDB-3kev: X-ray crystal structure of a DCUN1 domain-containing protein from... -

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Basic information

Entry
Database: PDB / ID: 3kev
TitleX-ray crystal structure of a DCUN1 domain-containing protein from Galdieria sulfuraria
ComponentsGalieria sulfuraria DCUN1 domain-containing protein
Keywordsstructural genomics / unknown function / cullin / neddylation / DCUN1 / DCN-1 / E3 / Center for Eukaryotic Structural Genomics / PSI / Protein Structure Initiative / CESG
Function / homology
Function and homology information


ubiquitin-like protein binding / protein neddylation / ubiquitin conjugating enzyme binding / cullin family protein binding / ubiquitin ligase complex
Similarity search - Function
Cullin, PONY binding domain / Potentiating neddylation domain / Defective-in-cullin neddylation protein / DCN1-like, PONY binding domain / Cullin binding / DCUN1 domain profile. / EF-hand / Recoverin; domain 1 / Orthogonal Bundle / Mainly Alpha
Similarity search - Domain/homology
ACETATE ION / Defective in cullin neddylation protein
Similarity search - Component
Biological speciesGaldieria sulphuraria (eukaryote)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MAD, molecular replacement / MAD / molecular replacement / Resolution: 1.3 Å
AuthorsBurgie, E.S. / Bingman, C.A. / Phillips Jr., G.N. / Center for Eukaryotic Structural Genomics (CESG)
CitationJournal: Proteins / Year: 2011
Title: Structural architecture of Galdieria sulphuraria DCN1L.
Authors: Burgie, E.S. / Bingman, C.A. / Makino, S. / Wesenberg, G.E. / Pan, X. / Fox, B.G. / Phillips, G.N.
History
DepositionOct 26, 2009Deposition site: RCSB / Processing site: RCSB
Revision 1.0Dec 1, 2009Provider: repository / Type: Initial release
Revision 1.1Jul 13, 2011Group: Version format compliance
Revision 1.2Nov 1, 2017Group: Refinement description / Category: software
Item: _software.classification / _software.contact_author ..._software.classification / _software.contact_author / _software.contact_author_email / _software.date / _software.language / _software.location / _software.name / _software.type / _software.version
Revision 1.3Oct 16, 2024Group: Data collection / Database references ...Data collection / Database references / Derived calculations / Structure summary
Category: chem_comp_atom / chem_comp_bond ...chem_comp_atom / chem_comp_bond / database_2 / pdbx_entry_details / pdbx_modification_feature / struct_conn / struct_site
Item: _database_2.pdbx_DOI / _database_2.pdbx_database_accession ..._database_2.pdbx_DOI / _database_2.pdbx_database_accession / _struct_conn.pdbx_leaving_atom_flag / _struct_site.pdbx_auth_asym_id / _struct_site.pdbx_auth_comp_id / _struct_site.pdbx_auth_seq_id

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: Galieria sulfuraria DCUN1 domain-containing protein
hetero molecules


Theoretical massNumber of molelcules
Total (without water)23,5394
Polymers23,3251
Non-polymers2143
Water3,171176
1


  • Idetical with deposited unit
  • defined by author&software
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
MethodPISA
Unit cell
Length a, b, c (Å)36.743, 52.422, 49.113
Angle α, β, γ (deg.)90.000, 111.170, 90.000
Int Tables number4
Space group name H-MP1211

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Components

#1: Protein Galieria sulfuraria DCUN1 domain-containing protein


Mass: 23325.146 Da / Num. of mol.: 1
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Galdieria sulphuraria (eukaryote) / Description: Wheat germ cell free / Plasmid: pEU-His-Flexi / Production host: CELL-FREE SYNTHESIS (others) / References: UniProt: D1MPT4*PLUS
#2: Chemical ChemComp-SO4 / SULFATE ION


Mass: 96.063 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: SO4
#3: Chemical ChemComp-ACT / ACETATE ION


Mass: 59.044 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: C2H3O2
#4: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 176 / Source method: isolated from a natural source / Formula: H2O
Has protein modificationY

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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

Crystal
IDDensity Matthews3/Da)Density % sol (%)
11.8934.95
2
Crystal grow
Temperature (K)Crystal-IDMethodpHDetails
2931vapor diffusion, sitting drop530% PEG 1500, 100 mM sodium acetate pH 5.0, 150 mM MgCl2, 50 mM NaCl, 5 mM HEPES pH 7.0, 0.3 mM TCEP, VAPOR DIFFUSION, SITTING DROP, temperature 293K
2932vapor diffusion, sitting drop527% PEG 1500, 100 mM sodium acetate pH 5.0, 150 mM LiCl2, 50 mM NaCl, 5 mM HEPES pH 7.0, 0.3 mM TCEP, VAPOR DIFFUSION, SITTING DROP, temperature 293K

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Data collection

Diffraction
IDMean temperature (K)Crystal-ID
11001
21002
Diffraction source
SourceSiteBeamlineIDWavelength (Å)
SYNCHROTRONAPS 21-ID-D10.97625
SYNCHROTRONAPS 23-ID-D20.96426, 0.97949, 0.97965, 0.99522
Detector
TypeIDDetectorDate
MARMOSAIC 300 mm CCD1CCDOct 12, 2009
MARMOSAIC 300 mm CCD2CCDOct 18, 2009
Radiation
IDMonochromatorProtocolScattering typeWavelength-ID
1Si(111)SINGLE WAVELENGTHx-ray1
2Si(111)MADx-ray1
Radiation wavelength
IDWavelength (Å)Relative weight
10.976251
20.964261
30.979491
40.979651
50.995221
ReflectionRedundancy: 7.5 % / Av σ(I) over netI: 23.15 / Number: 112798 / Rmerge(I) obs: 0.075 / Χ2: 1.24 / D res high: 1.81 Å / D res low: 50 Å / Num. obs: 15091 / % possible obs: 97.7
Diffraction reflection shell
Highest resolution (Å)Lowest resolution (Å)% possible obs (%)IDRmerge(I) obsChi squaredRedundancy
4.915097.510.081.3327.3
3.94.9199.210.0460.887.5
3.413.99910.0410.8927.3
3.093.4198.810.0430.9127.3
2.873.099910.0460.9447.4
2.72.879910.0531.0497.4
2.572.798.410.0611.1837.5
2.462.5798.310.0731.2627.6
2.362.4698.210.0761.317.7
2.282.3698.210.0761.2347.7
2.212.2899.210.0851.2897.6
2.152.2197.210.0941.2957.7
2.092.1597.710.1051.267.7
2.042.0998.310.1121.3467.7
1.992.0496.910.1311.3877.7
1.951.9998.810.1321.3967.6
1.911.9596.110.161.467.6
1.871.9198.410.1851.447.5
1.841.879610.1921.4087.4
1.811.8489.710.2311.5876.3
ReflectionResolution: 1.3→50 Å / Num. all: 42886 / Num. obs: 42243 / % possible obs: 98.5 % / Redundancy: 4 % / Rmerge(I) obs: 0.04 / Χ2: 1.012 / Net I/σ(I): 16
Reflection shellResolution: 1.3→1.32 Å / Redundancy: 3 % / Rmerge(I) obs: 0.388 / Mean I/σ(I) obs: 2.6 / Num. unique all: 1849 / Χ2: 1.114 / % possible all: 87.2

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Phasing

Phasing
Method
MAD
molecular replacement
Phasing MAD set
IDR cullis acentricR cullis centricHighest resolution (Å)Lowest resolution (Å)Reflection acentricReflection centric
ISO_1001.7934.2214383695
ISO_20.4810.4881.7934.2214378692
ISO_30.7310.7341.7934.2213794660
ISO_40.6420.671.7934.2214360692
ANO_10.95801.7934.22143230
ANO_20.97401.7934.22149500
ANO_30.99901.7934.22137420
ANO_40.99401.7934.22144180
Phasing MAD set shell
IDResolution (Å)R cullis acentricR cullis centricReflection acentricReflection centric
ISO_17.81-34.220015834
ISO_15.6-7.810030439
ISO_14.59-5.60039837
ISO_13.98-4.590046939
ISO_13.57-3.980053537
ISO_13.26-3.570059140
ISO_13.02-3.260063734
ISO_12.83-3.020070835
ISO_12.67-2.830073539
ISO_12.53-2.670077434
ISO_12.41-2.530081735
ISO_12.31-2.410087137
ISO_12.22-2.310089936
ISO_12.14-2.220092231
ISO_12.07-2.140096133
ISO_12-2.070099238
ISO_11.94-200101731
ISO_11.89-1.9400105841
ISO_11.84-1.8900107526
ISO_11.79-1.840046219
ANO_17.81-34.220.76901570
ANO_15.6-7.810.6603040
ANO_14.59-5.60.80703980
ANO_13.98-4.590.86804690
ANO_13.57-3.980.87205310
ANO_13.26-3.570.89405870
ANO_13.02-3.260.85806330
ANO_12.83-3.020.8807060
ANO_12.67-2.830.88807330
ANO_12.53-2.670.92307720
ANO_12.41-2.530.93708170
ANO_12.31-2.410.96508710
ANO_12.22-2.310.97308980
ANO_12.14-2.220.98409220
ANO_12.07-2.140.9909590
ANO_12-2.070.99209920
ANO_11.94-20.995010160
ANO_11.89-1.940.997010570
ANO_11.84-1.890.998010720
ANO_11.79-1.840.99904290
ISO_27.81-34.220.570.87115534
ISO_25.6-7.810.430.53730439
ISO_24.59-5.60.4590.4339837
ISO_23.98-4.590.4170.38946939
ISO_23.57-3.980.4450.27553537
ISO_23.26-3.570.3590.39859140
ISO_23.02-3.260.3290.39963734
ISO_22.83-3.020.3390.33370835
ISO_22.67-2.830.3570.43273539
ISO_22.53-2.670.3530.40677434
ISO_22.41-2.530.3550.481735
ISO_22.31-2.410.40.41487137
ISO_22.22-2.310.4060.5889936
ISO_22.14-2.220.4550.5792231
ISO_22.07-2.140.4870.61996133
ISO_22-2.070.5160.6699238
ISO_21.94-20.530.57101731
ISO_21.89-1.940.5770.551105841
ISO_21.84-1.890.6340.65107424
ISO_21.79-1.840.5990.45846118
ANO_27.81-34.220.78601520
ANO_25.6-7.810.67403040
ANO_24.59-5.60.80803980
ANO_23.98-4.590.88704690
ANO_23.57-3.980.88105350
ANO_23.26-3.570.88405900
ANO_23.02-3.260.89106400
ANO_22.83-3.020.90107080
ANO_22.67-2.830.90707350
ANO_22.53-2.670.94407740
ANO_22.41-2.530.95708170
ANO_22.31-2.410.97708700
ANO_22.22-2.310.98208990
ANO_22.14-2.220.9909230
ANO_22.07-2.140.99409630
ANO_22-2.070.99609920
ANO_21.94-20.997010160
ANO_21.89-1.940.998010580
ANO_21.84-1.890.999010750
ANO_21.79-1.841.001010320
ISO_37.81-34.220.770.84114433
ISO_35.6-7.810.7430.77830438
ISO_34.59-5.60.7250.67739837
ISO_33.98-4.590.70.69346939
ISO_33.57-3.980.7070.67353537
ISO_33.26-3.570.7060.73159040
ISO_33.02-3.260.7040.70163734
ISO_32.83-3.020.7130.67570835
ISO_32.67-2.830.7020.75273539
ISO_32.53-2.670.710.80377434
ISO_32.41-2.530.7050.75181735
ISO_32.31-2.410.7180.6987037
ISO_32.22-2.310.7130.77789936
ISO_32.14-2.220.7350.80292231
ISO_32.07-2.140.7460.72196133
ISO_32-2.070.760.83399238
ISO_31.94-20.7710.713101729
ISO_31.89-1.940.7960.849105038
ISO_31.84-1.890.8550.71197016
ISO_31.79-1.841.3760.96921
ANO_37.81-34.220.96901420
ANO_35.6-7.810.94803040
ANO_34.59-5.60.97203970
ANO_33.98-4.590.98204690
ANO_33.57-3.980.98905340
ANO_33.26-3.570.98305890
ANO_33.02-3.260.98906410
ANO_32.83-3.020.99807080
ANO_32.67-2.830.99907360
ANO_32.53-2.670.99907740
ANO_32.41-2.530.99908170
ANO_32.31-2.410.99908700
ANO_32.22-2.310.99908990
ANO_32.14-2.22109210
ANO_32.07-2.14109600
ANO_32-2.07109910
ANO_31.94-21010160
ANO_31.89-1.941010440
ANO_31.84-1.89109290
ANO_31.79-1.841010
ISO_47.81-34.220.7770.86314132
ISO_45.6-7.810.7130.77630439
ISO_44.59-5.60.660.60739837
ISO_43.98-4.590.5990.62146839
ISO_43.57-3.980.5970.59253537
ISO_43.26-3.570.5930.64658940
ISO_43.02-3.260.5950.6163734
ISO_42.83-3.020.6130.54170835
ISO_42.67-2.830.60.6573539
ISO_42.53-2.670.6150.73477434
ISO_42.41-2.530.6110.61581735
ISO_42.31-2.410.6240.61187137
ISO_42.22-2.310.6110.64889936
ISO_42.14-2.220.6430.75192231
ISO_42.07-2.140.6470.65496133
ISO_42-2.070.6650.73299238
ISO_41.94-20.6710.654101731
ISO_41.89-1.940.7030.743105741
ISO_41.84-1.890.7670.785107526
ISO_41.79-1.840.8160.73746018
ANO_47.81-34.220.89701380
ANO_45.6-7.810.77103040
ANO_44.59-5.60.90103960
ANO_43.98-4.590.93904660
ANO_43.57-3.980.93605340
ANO_43.26-3.570.94405880
ANO_43.02-3.260.95806400
ANO_42.83-3.020.95507090
ANO_42.67-2.830.98707350
ANO_42.53-2.670.98207740
ANO_42.41-2.530.98908170
ANO_42.31-2.410.9908710
ANO_42.22-2.310.99608990
ANO_42.14-2.220.99709220
ANO_42.07-2.140.99809630
ANO_42-2.070.99909920
ANO_41.94-20.999010170
ANO_41.89-1.941010570
ANO_41.84-1.891010680
ANO_41.79-1.84105280
Phasing MAD set siteCartn x: 5.162 Å / Cartn y: 0.052 Å / Cartn z: 12.191 Å / Atom type symbol: SE / B iso: 36.84 / Occupancy: 0.83
Phasing MR
Highest resolutionLowest resolution
Rotation1.6 Å20.99 Å
Translation1.6 Å20.99 Å
Phasing dmMethod: Solvent flattening and Histogram matching / Reflection: 15703
Phasing dm shell
Resolution (Å)Delta phi finalFOM Reflection
5.72-100540.782504
4.53-5.7247.60.915502
3.94-4.5354.50.914515
3.59-3.9455.60.914506
3.32-3.5960.40.878503
3.13-3.3271.60.878504
2.97-3.1362.40.861507
2.84-2.9770.70.882516
2.72-2.8463.50.875519
2.62-2.7268.70.881532
2.53-2.6263.70.876572
2.45-2.5369.60.877573
2.38-2.4566.70.865608
2.31-2.3867.70.876620
2.24-2.3170.40.86620
2.19-2.2471.50.875652
2.13-2.1970.20.886666
2.08-2.1373.20.865679
2.04-2.0875.10.858685
1.99-2.0475.20.867726
1.95-1.9976.60.848733
1.91-1.9580.30.833694
1.88-1.9181.80.81792
1.84-1.8885.40.791730
1.79-1.8487.20.7431245

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Processing

Software
NameVersionClassificationNB
DENZOdata reduction
SCALEPACKdata scaling
MOLREPphasing
SHARPphasing
DM6.1phasing
PHENIX1.5_2refinement
PDB_EXTRACT3.005data extraction
HKL-2000data collection
HKL-2000data scaling
RefinementMethod to determine structure: MAD, molecular replacement / Resolution: 1.3→20.984 Å / Occupancy max: 1.4 / Occupancy min: 0.19 / FOM work R set: 0.922 / SU ML: 0.16 / σ(F): 1.35 / Stereochemistry target values: ML
RfactorNum. reflection% reflectionSelection details
Rfree0.178 2111 5.01 %random
Rwork0.155 ---
all0.156 42820 --
obs0.156 42161 98.46 %-
Solvent computationShrinkage radii: 0.9 Å / VDW probe radii: 1.11 Å / Solvent model: FLAT BULK SOLVENT MODEL / Bsol: 60.058 Å2 / ksol: 0.4 e/Å3
Displacement parametersBiso max: 52.77 Å2 / Biso mean: 20.012 Å2 / Biso min: 7.22 Å2
Baniso -1Baniso -2Baniso -3
1--1.285 Å2-0 Å22.593 Å2
2---2.06 Å2-0 Å2
3---3.345 Å2
Refinement stepCycle: LAST / Resolution: 1.3→20.984 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms1525 0 13 176 1714
Refine LS restraints
Refine-IDTypeDev idealNumber
X-RAY DIFFRACTIONf_bond_d0.0181800
X-RAY DIFFRACTIONf_angle_d1.5662465
X-RAY DIFFRACTIONf_chiral_restr0.101259
X-RAY DIFFRACTIONf_plane_restr0.007328
X-RAY DIFFRACTIONf_dihedral_angle_d17.426679
LS refinement shell

Refine-ID: X-RAY DIFFRACTION / Total num. of bins used: 15

Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkNum. reflection all% reflection obs (%)
1.3-1.330.1971240.1832388251288
1.33-1.3630.211430.1732599274296
1.363-1.40.21480.1562640278899
1.4-1.4420.1941410.14726872828100
1.442-1.4880.1921260.13427412867100
1.488-1.5410.1631610.12226692830100
1.541-1.6030.161390.11627172856100
1.603-1.6760.1781370.11327092846100
1.676-1.7640.1251370.11626962833100
1.764-1.8750.1721440.11827102854100
1.875-2.0190.1531330.1327062839100
2.019-2.2220.1431380.12527232861100
2.222-2.5430.1781390.14227182857100
2.543-3.2030.1861510.15927292880100
3.203-20.9870.1711500.1692618276894

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