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- PDB-3guc: Human Ubiquitin-activating Enzyme 5 in Complex with AMPPNP -

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Basic information

Entry
Database: PDB / ID: 3guc
TitleHuman Ubiquitin-activating Enzyme 5 in Complex with AMPPNP
ComponentsUbiquitin-like modifier-activating enzyme 5
KeywordsTRANSFERASE / ROSSMANN FOLD / ATP-BINDING / UBL CONJUGATION PATHWAY / Structural Genomics / Structural Genomics Consortium / SGC
Function / homology
Function and homology information


UFM1 activating enzyme activity / protein ufmylation / protein K69-linked ufmylation / megakaryocyte differentiation / regulation of intracellular estrogen receptor signaling pathway / reticulophagy / neuromuscular process / localization / response to endoplasmic reticulum stress / erythrocyte differentiation ...UFM1 activating enzyme activity / protein ufmylation / protein K69-linked ufmylation / megakaryocyte differentiation / regulation of intracellular estrogen receptor signaling pathway / reticulophagy / neuromuscular process / localization / response to endoplasmic reticulum stress / erythrocyte differentiation / Antigen processing: Ubiquitination & Proteasome degradation / intracellular membrane-bounded organelle / endoplasmic reticulum membrane / Golgi apparatus / protein homodimerization activity / zinc ion binding / ATP binding / nucleus / cytosol / cytoplasm
Similarity search - Function
ThiF/MoeB/HesA family / Ubiquitin-activating enzyme / THIF-type NAD/FAD binding fold / ThiF family / D-isomer specific 2-hydroxyacid dehydrogenase, NAD-binding domain conserved site 1 / NAD(P)-binding Rossmann-like Domain / Rossmann fold / 3-Layer(aba) Sandwich / Alpha Beta
Similarity search - Domain/homology
PHOSPHOAMINOPHOSPHONIC ACID-ADENYLATE ESTER / Ubiquitin-like modifier-activating enzyme 5
Similarity search - Component
Biological speciesHomo sapiens (human)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 2.25 Å
AuthorsWalker, J.R. / Bacik, J.P. / Li, Y. / Weigelt, J. / Bountra, C. / Arrowsmith, C.H. / Edwards, A.M. / Bochkarev, A. / Dhe-Paganon, S. / Structural Genomics Consortium (SGC)
CitationJournal: To be Published
Title: Human Ubiquitin-activating Enzyme 5 in Complex with AMPPNP
Authors: Walker, J.R. / Bacik, J.P. / Li, Y. / Weigelt, J. / Bountra, C. / Arrowsmith, C.H. / Edwards, A.M. / Bochkarev, A. / Dhe-Paganon, S.
History
DepositionMar 29, 2009Deposition site: RCSB / Processing site: RCSB
Revision 1.0May 26, 2009Provider: repository / Type: Initial release
Revision 1.1Jul 13, 2011Group: Version format compliance
Revision 1.2Nov 12, 2014Group: Structure summary
Revision 1.3Sep 6, 2023Group: Data collection / Database references ...Data collection / Database references / Derived calculations / Refinement description
Category: chem_comp_atom / chem_comp_bond ...chem_comp_atom / chem_comp_bond / database_2 / pdbx_initial_refinement_model / struct_site
Item: _database_2.pdbx_DOI / _database_2.pdbx_database_accession ..._database_2.pdbx_DOI / _database_2.pdbx_database_accession / _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

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Assembly

Deposited unit
A: Ubiquitin-like modifier-activating enzyme 5
B: Ubiquitin-like modifier-activating enzyme 5
hetero molecules


Theoretical massNumber of molelcules
Total (without water)64,9695
Polymers64,3322
Non-polymers6373
Water2,252125
1


  • Idetical with deposited unit
  • defined by author&software
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area3150 Å2
ΔGint-19 kcal/mol
Surface area20900 Å2
MethodPISA
Unit cell
Length a, b, c (Å)77.394, 77.394, 205.892
Angle α, β, γ (deg.)90.00, 90.00, 120.00
Int Tables number154
Space group name H-MP3221

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Components

#1: Protein Ubiquitin-like modifier-activating enzyme 5 / Ubiquitin-activating enzyme 5 / Ubiquitin-activating enzyme E1 domain-containing protein 1 / UFM1- ...Ubiquitin-activating enzyme 5 / Ubiquitin-activating enzyme E1 domain-containing protein 1 / UFM1-activating enzyme / ThiFP1


Mass: 32165.947 Da / Num. of mol.: 2
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Homo sapiens (human) / Gene: UBA5, UBE1DC1 / Plasmid: PET28A-LIC / Production host: Escherichia coli BL21(DE3) (bacteria) / Strain (production host): BL21 (DE3)
References: UniProt: Q9GZZ9, Transferases; Transferring phosphorus-containing groups; Nucleotidyltransferases
#2: Chemical ChemComp-ZN / ZINC ION


Mass: 65.409 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: Zn
#3: Chemical ChemComp-ANP / PHOSPHOAMINOPHOSPHONIC ACID-ADENYLATE ESTER


Mass: 506.196 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: C10H17N6O12P3 / Comment: AMP-PNP, energy-carrying molecule analogue*YM
#4: Water ChemComp-HOH / water / Water


Mass: 18.015 Da / Num. of mol.: 125 / 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: 2.51 Å3/Da / Density % sol: 51.05 %
Crystal growTemperature: 287 K / pH: 6.2
Details: 1 M LITHIUM SULPHATE, 0.3 M AMMONIUM SULPHATE,0.1 M SODIUM CITRATE, PH 6.2, VAPOR DIFFUSION, HANGING DROP, TEMPERATURE 287K

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: APS / Beamline: 19-ID / Wavelength: 0.97935
DetectorType: ADSC QUANTUM 315 / Detector: CCD / Date: Mar 18, 2009
RadiationMonochromator: SI(111) / Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.97935 Å / Relative weight: 1
ReflectionResolution: 2.25→50 Å / Num. obs: 34922 / % possible obs: 100 % / Observed criterion σ(I): -3 / Redundancy: 10.5 % / Rsym value: 0.131 / Net I/σ(I): 23.878
Reflection shellResolution: 2.25→2.29 Å / Redundancy: 8.9 % / Mean I/σ(I) obs: 2.89 / Rsym value: 0.838 / % possible all: 100

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Processing

Software
NameVersionClassification
HKL-3000data collection
PHASERphasing
REFMAC5.5.0063refinement
HKL-3000data reduction
HKL-3000data scaling
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: 1ZFN
Resolution: 2.25→40.82 Å / Cor.coef. Fo:Fc: 0.936 / Cor.coef. Fo:Fc free: 0.9 / SU B: 10.761 / SU ML: 0.128 / Cross valid method: THROUGHOUT / ESU R: 0.208 / ESU R Free: 0.195 / Stereochemistry target values: MAXIMUM LIKELIHOOD
Details: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS. ATOM RECORD CONTAINS SUM OF TLS AND RESIDUAL B FACTORS, ANISOU RECORD CONTAINS SUM OF TLS AND RESIDUAL U FACTORS.
RfactorNum. reflection% reflectionSelection details
Rfree0.25485 1770 5.1 %RANDOM
Rwork0.20303 ---
obs0.20562 33065 99.97 %-
all-34835 --
Solvent computationIon probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.4 Å / Solvent model: MASK
Displacement parametersBiso mean: 55.853 Å2
Baniso -1Baniso -2Baniso -3
1-0.96 Å20.48 Å20 Å2
2--0.96 Å20 Å2
3----1.45 Å2
Refinement stepCycle: LAST / Resolution: 2.25→40.82 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms3525 0 29 128 3682
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.0150.0223624
X-RAY DIFFRACTIONr_bond_other_d
X-RAY DIFFRACTIONr_angle_refined_deg1.5311.9624907
X-RAY DIFFRACTIONr_angle_other_deg
X-RAY DIFFRACTIONr_dihedral_angle_1_deg5.5945458
X-RAY DIFFRACTIONr_dihedral_angle_2_deg38.30225.38158
X-RAY DIFFRACTIONr_dihedral_angle_3_deg18.22415590
X-RAY DIFFRACTIONr_dihedral_angle_4_deg25.1161513
X-RAY DIFFRACTIONr_chiral_restr0.120.2567
X-RAY DIFFRACTIONr_gen_planes_refined0.0060.0212713
X-RAY DIFFRACTIONr_gen_planes_other
X-RAY DIFFRACTIONr_nbd_refined
X-RAY DIFFRACTIONr_nbd_other
X-RAY DIFFRACTIONr_nbtor_refined
X-RAY DIFFRACTIONr_nbtor_other
X-RAY DIFFRACTIONr_xyhbond_nbd_refined
X-RAY DIFFRACTIONr_xyhbond_nbd_other
X-RAY DIFFRACTIONr_metal_ion_refined
X-RAY DIFFRACTIONr_metal_ion_other
X-RAY DIFFRACTIONr_symmetry_vdw_refined
X-RAY DIFFRACTIONr_symmetry_vdw_other
X-RAY DIFFRACTIONr_symmetry_hbond_refined
X-RAY DIFFRACTIONr_symmetry_hbond_other
X-RAY DIFFRACTIONr_symmetry_metal_ion_refined
X-RAY DIFFRACTIONr_symmetry_metal_ion_other
X-RAY DIFFRACTIONr_mcbond_it1.15722297
X-RAY DIFFRACTIONr_mcbond_other
X-RAY DIFFRACTIONr_mcangle_it1.96733697
X-RAY DIFFRACTIONr_scbond_it3.26241327
X-RAY DIFFRACTIONr_scangle_it4.25251210
X-RAY DIFFRACTIONr_rigid_bond_restr
X-RAY DIFFRACTIONr_sphericity_free
X-RAY DIFFRACTIONr_sphericity_bonded
LS refinement shellResolution: 2.25→2.308 Å / Total num. of bins used: 20
RfactorNum. reflection% reflection
Rfree0.34 128 -
Rwork0.248 2392 -
obs--99.92 %
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)
19.4028-2.19372.14934.16281.58465.17270.0606-0.945-0.40680.4514-0.03290.52460.1163-0.2544-0.02770.23650.0243-0.00120.18780.0110.1902-44.45262.4502-4.6377
22.2155-1.3552-1.28551.6123-0.62923.31040.0209-0.03460.23810.0934-0.0566-0.2514-0.19990.19120.03570.18080.0625-0.00320.1873-0.04430.124-39.509116.9882-11.5278
31.2067-0.0286-0.29373.1862-0.62094.51060.0893-0.06530.16050.23780.0296-0.16560.0710.1873-0.11890.15750.0752-0.01390.2159-0.02940.1189-37.436412.4353-9.7183
46.32682.30624.09621.13252.58567.4142-0.0594-0.24910.27920.03380.0225-0.0379-0.28140.46480.03690.5299-0.0009-0.10250.3604-0.00560.5199-26.664224.3203-16.4518
51.6060.5658-2.97139.94961.81376.33690.2192-0.6061-0.09440.3074-0.1419-0.8886-0.35011.1499-0.07730.3326-0.0238-0.04780.45130.00690.3878-23.99221.0181-15.4136
63.51822.6777-3.86247.5317-3.42564.3195-0.1043-0.2983-0.10040.0098-0.0093-0.70830.16870.3750.11360.20710.068-0.05950.3371-0.03040.245-28.033412.5096-9.5697
70.632-0.4981-0.132.7809-1.67843.8015-0.01690.04680.0234-0.065-0.1429-0.42630.02420.40840.15980.17920.08520.00450.1877-0.01680.1169-35.911613.4486-23.2923
83.7753-0.4259-1.10347.8033-3.57735.3577-0.069-0.1646-0.2479-0.16570.08050.06860.3607-0.1763-0.01150.21160.1093-0.01150.1274-0.03060.1179-42.63815.4984-22.3318
91.7128-0.35080.90910.5602-0.0433.0064-0.06630.0940.14250.04370.0484-0.3559-0.24090.07520.01790.18490.06660.01040.1654-0.02850.2411-44.325519.7556-18.6269
106.06521.7275-0.16945.0748-0.5284.15830.20750.15010.0097-0.3853-0.2577-0.06670.11780.09480.05020.15430.106-0.00770.0993-0.03840.0521-49.295418.1092-23.4638
110.75421.23520.2785.39863.70194.83680.00940.03620.1438-0.5260.116-0.1925-0.56810.3788-0.12550.18030.0346-0.01320.2029-0.01780.197-44.511631.0452-10.5459
121.4251-0.3701-0.0641.9625-1.0991.6235-0.035-0.02890.0177-0.0999-0.03460.0847-0.0502-0.1830.06960.14730.065-0.01650.1806-0.00520.1158-54.745524.6664-17.3571
134.41784.29464.79734.25314.67095.2443-0.9926-0.06451.027-0.8858-0.04910.7986-1.17580.02991.04170.8684-0.0232-0.11550.5654-0.04730.923-41.021142.2828-4.9768
148.60526.88070.00596.26081.51986.04750.1681-0.10410.0238-0.069-0.0138-0.3751-0.34240.5728-0.15430.22130.04640.05130.2420.00010.3715-39.109730.6643-2.9506
151.2255-0.85610.230.85740.53912.1961-0.055-0.0926-0.03770.01860.0110.08710.0561-0.0390.0440.14990.0278-0.00050.1828-0.02030.1288-51.481722.1495-4.2418
162.25932.38515.66742.63225.919414.629-0.06890.02010.1508-0.0701-0.0220.1755-0.1894-0.01780.0910.18910.0181-0.00020.21020.01150.2329-61.615224.9689-13.8722
174.0365-0.8143-0.59497.59111.65481.97610.14720.43240.0566-0.4811-0.1491-0.08180.0066-0.090.00190.18660.0208-0.02730.1989-0.00380.1309-59.923520.3884-29.2275
185.15861.1532-0.57574.2123-0.64730.4360.08170.4676-0.8840.1965-0.0991.07930.1524-0.47510.01730.4346-0.201-0.05970.7371-0.30860.6641-66.239112.6501-24.0769
196.0677-0.0827-4.44897.05080.18476.76970.14910.81330.6279-0.87640.0508-0.0263-0.33710.1802-0.19990.29570.0070.02340.33160.06010.2741-30.151741.77976.9204
203.3559-0.21560.2590.7464-1.39482.61410.14110.1741-0.3242-0.1076-0.05110.05380.20310.106-0.08990.1020.0418-0.02240.2532-0.04260.1331-35.449925.383213.0396
215.84161.92711.29847.43370.78144.4906-0.09360.4972-0.1489-0.50980.02410.0505-0.07970.00470.06950.06280.0233-0.01090.251-0.02420.0798-30.391230.34547.0356
228.6503-0.7516-4.73710.41830.05665.278-0.2666-0.114-1.02870.05040.03970.04890.78280.02890.2270.2133-0.0114-0.05530.2941-0.01590.2298-29.518721.02219.8589
231.8770.022-1.1871.0928-0.06144.4312-0.0473-0.0301-0.16560.0301-0.0627-0.14550.11750.3450.110.00990.013-0.02190.274-0.020.1266-28.159527.864820.4871
245.46691.61662.14133.40752.5223.79990.1683-0.15230.10960.0077-0.2076-0.3193-0.19780.09940.03930.16910.0017-0.00960.27320.01170.1724-33.622934.817518.3681
252.2471-1.4591-0.99467.08770.3114.0489-0.008-0.2778-0.3083-0.12470.0939-0.09780.3356-0.0646-0.08590.11910.0156-0.02690.29840.00910.1204-43.870326.246625.6896
265.1453-0.5253-0.93673.22920.70620.39140.0105-0.47310.14010.1815-0.0472-0.22620.02530.12150.03670.1745-0.0278-0.03550.2598-0.02270.1441-41.648636.787524.1238
272.1963.37221.59946.33352.38885.0960.31-0.2712-0.30220.0289-0.4913-0.13090.7334-0.41120.18140.2985-0.0083-0.0750.2613-0.00010.2454-47.273820.436512.9411
281.60571.11020.47288.93273.4462.78250.0452-0.0269-0.03670.1333-0.10180.02550.1191-0.11720.05650.13330.01010.0040.1810.01340.1384-49.020428.919413.5216
294.10871.21520.68870.7989-0.7492.55580.0735-0.0401-0.1252-0.0951-0.07460.04250.1373-0.02030.00110.14840.0729-0.02880.2375-0.02490.1875-54.330830.413219.8298
307.3697-6.39720.69656.6866-0.31080.14650.44050.101-0.7551-0.134-0.25940.1720.1423-0.0665-0.18110.5109-0.12-0.0270.4472-0.03970.3341-48.339912.18264.5225
313.98322.63961.27148.87461.14613.12530.07360.0554-0.1881-0.2753-0.0813-0.10830.09260.02780.00770.08210.035-0.0010.1988-0.01210.0948-43.249925.31185.1092
325.50780.91893.79318.88491.34264.7576-0.00770.17240.2781-0.1714-0.0794-0.0178-0.4670.02740.08710.1289-0.00880.0020.2466-0.01310.1166-40.357639.32087.3272
331.5493-0.9729-0.02420.61410.07941.69830.01780.02370.0895-0.0233-0.0163-0.0476-0.17890.061-0.00150.13840.0067-0.00580.1944-0.02470.1317-50.102232.76087.3908
341.95820.3062-0.50429.00654.14333.3473-0.01060.0118-0.1592-0.1392-0.08550.07940.0748-0.18120.09610.13440.0284-0.02060.2209-0.01380.1452-56.446424.53084.0898
353.86640.2520.63644.25811.08512.2874-0.0324-0.56410.03150.45950.0678-0.0015-0.2052-0.0815-0.03540.12770.03860.00420.2822-0.02830.1004-53.12337.221728.7626
365.4670.4534-2.10198.83090.72013.39240.0873-0.49070.9953-0.3024-0.0491-0.1806-0.811-0.2628-0.03820.39090.0503-0.03110.2931-0.14170.2839-49.844347.477226.9155
Refinement TLS group
IDRefine-IDRefine TLS-IDAuth asym-IDAuth seq-ID
1X-RAY DIFFRACTION1A69 - 73
2X-RAY DIFFRACTION2A74 - 89
3X-RAY DIFFRACTION3A90 - 104
4X-RAY DIFFRACTION4A105 - 121
5X-RAY DIFFRACTION5A122 - 127
6X-RAY DIFFRACTION6A128 - 139
7X-RAY DIFFRACTION7A140 - 159
8X-RAY DIFFRACTION8A160 - 173
9X-RAY DIFFRACTION9A174 - 186
10X-RAY DIFFRACTION10A187 - 202
11X-RAY DIFFRACTION11A203 - 214
12X-RAY DIFFRACTION12A215 - 235
13X-RAY DIFFRACTION13A236 - 251
14X-RAY DIFFRACTION14A252 - 260
15X-RAY DIFFRACTION15A261 - 293
16X-RAY DIFFRACTION16A294 - 299
17X-RAY DIFFRACTION17A300 - 311
18X-RAY DIFFRACTION18A312 - 329
19X-RAY DIFFRACTION19B69 - 74
20X-RAY DIFFRACTION20B75 - 88
21X-RAY DIFFRACTION21B89 - 100
22X-RAY DIFFRACTION22B101 - 128
23X-RAY DIFFRACTION23B129 - 168
24X-RAY DIFFRACTION24B169 - 183
25X-RAY DIFFRACTION25B184 - 195
26X-RAY DIFFRACTION26B196 - 202
27X-RAY DIFFRACTION27B203 - 211
28X-RAY DIFFRACTION28B212 - 220
29X-RAY DIFFRACTION29B221 - 237
30X-RAY DIFFRACTION30B238 - 255
31X-RAY DIFFRACTION31B256 - 267
32X-RAY DIFFRACTION32B268 - 275
33X-RAY DIFFRACTION33B276 - 286
34X-RAY DIFFRACTION34B287 - 295
35X-RAY DIFFRACTION35B296 - 310
36X-RAY DIFFRACTION36B311 - 319

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