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- PDB-2jk0: Structural and functional insights into Erwinia carotovora L- asp... -

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Basic information

Entry
Database: PDB / ID: 2jk0
TitleStructural and functional insights into Erwinia carotovora L- asparaginase
ComponentsL-ASPARAGINASE
KeywordsHYDROLASE / ERWINIA / ENZYME THERAPY / PROTEIN STABILITY / LEUKEMIA TREATMENT
Function / homology
Function and homology information


asparagine metabolic process / asparaginase / asparaginase activity / cytosol
Similarity search - Function
L-asparaginase, N-terminal domain / Rossmann fold - #40 / L-asparaginase, type II / Asparaginase/glutaminase, active site 1 / Asparaginase / glutaminase active site signature 1. / L-asparaginase, C-terminal / Asparaginase/glutaminase, active site 2 / Asparaginase/glutaminase, C-terminal / Glutaminase/Asparaginase C-terminal domain / Asparaginase / glutaminase active site signature 2. ...L-asparaginase, N-terminal domain / Rossmann fold - #40 / L-asparaginase, type II / Asparaginase/glutaminase, active site 1 / Asparaginase / glutaminase active site signature 1. / L-asparaginase, C-terminal / Asparaginase/glutaminase, active site 2 / Asparaginase/glutaminase, C-terminal / Glutaminase/Asparaginase C-terminal domain / Asparaginase / glutaminase active site signature 2. / Asparaginase / Asparaginase/glutaminase-like / L-asparaginase, N-terminal / Asparaginase/glutaminase-like superfamily / L-asparaginase, N-terminal domain superfamily / Asparaginase, N-terminal / Asparaginase / glutaminase domain profile. / Rossmann fold / 3-Layer(aba) Sandwich / Alpha Beta
Similarity search - Domain/homology
ASPARTIC ACID / L-asparaginase
Similarity search - Component
Biological speciesPECTOBACTERIUM CAROTOVORUM (bacteria)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 2.5 Å
AuthorsPapageorgiou, A.C. / Posypanova, G.A. / Andersson, C.S. / Sokolov, N.N. / Krasotkina, J.
Citation
Journal: FEBS J. / Year: 2008
Title: Structural and Functional Insights Into Erwinia Carotovora L-Asparaginase.
Authors: Papageorgiou, A.C. / Posypanova, G.A. / Andersson, C.S. / Sokolov, N.N. / Krasotkina, J.
#1: Journal: Acta Crystallogr.,Sect.F / Year: 2005
Title: Crystallisation and Preliminary Crystallographic Analysis of L-Asparaginase from Erwinia Carotovora
Authors: E Wikman, L. / Krasotkina, J. / Kuchumova, A. / Sokolov, N.N. / Papageorgiou, A.C.
History
DepositionMay 23, 2008Deposition site: PDBE / Processing site: PDBE
Revision 1.0Aug 5, 2008Provider: repository / Type: Initial release
Revision 1.1Jul 13, 2011Group: Advisory / Version format compliance
Revision 1.2Jul 24, 2019Group: Data collection / Category: diffrn_source / Item: _diffrn_source.pdbx_synchrotron_site
Revision 1.3Dec 13, 2023Group: Data collection / Database references ...Data collection / Database references / Other / Refinement description
Category: chem_comp_atom / chem_comp_bond ...chem_comp_atom / chem_comp_bond / database_2 / pdbx_database_status / pdbx_initial_refinement_model
Item: _database_2.pdbx_DOI / _database_2.pdbx_database_accession / _pdbx_database_status.status_code_sf

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: L-ASPARAGINASE
B: L-ASPARAGINASE
C: L-ASPARAGINASE
D: L-ASPARAGINASE
E: L-ASPARAGINASE
F: L-ASPARAGINASE
G: L-ASPARAGINASE
H: L-ASPARAGINASE
hetero molecules


Theoretical massNumber of molelcules
Total (without water)275,68916
Polymers274,6258
Non-polymers1,0658
Water12,755708
1
A: L-ASPARAGINASE
B: L-ASPARAGINASE
C: L-ASPARAGINASE
D: L-ASPARAGINASE
hetero molecules


Theoretical massNumber of molelcules
Total (without water)137,8458
Polymers137,3124
Non-polymers5324
Water724
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area19700 Å2
ΔGint-63.5 kcal/mol
Surface area48290 Å2
MethodPQS
2
E: L-ASPARAGINASE
F: L-ASPARAGINASE
G: L-ASPARAGINASE
H: L-ASPARAGINASE
hetero molecules


Theoretical massNumber of molelcules
Total (without water)137,8458
Polymers137,3124
Non-polymers5324
Water724
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area17970 Å2
ΔGint-77.7 kcal/mol
Surface area48650 Å2
MethodPQS
Unit cell
Length a, b, c (Å)73.650, 135.630, 250.100
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
L-ASPARAGINASE


Mass: 34328.074 Da / Num. of mol.: 8
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) PECTOBACTERIUM CAROTOVORUM (bacteria) / Plasmid: ANSB1 / Production host: ESCHERICHIA COLI (E. coli) / Strain (production host): BL21(DE3) / References: UniProt: Q6Q4F4, asparaginase
#2: Chemical
ChemComp-ASP / ASPARTIC ACID


Type: L-peptide linking / Mass: 133.103 Da / Num. of mol.: 8 / Source method: obtained synthetically / Formula: C4H7NO4
#3: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 708 / Source method: isolated from a natural source / Formula: H2O

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Experimental details

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Experiment

ExperimentMethod: X-RAY DIFFRACTION

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Sample preparation

CrystalDensity Matthews: 2.29 Å3/Da / Density % sol: 45.82 % / Description: NONE

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: EMBL/DESY, HAMBURG / Beamline: X11 / Wavelength: 0.81
DetectorType: MARRESEARCH / Detector: CCD
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.81 Å / Relative weight: 1
ReflectionResolution: 2.5→20 Å / Num. obs: 83599 / % possible obs: 95.8 % / Observed criterion σ(I): -3 / Redundancy: 4.2 % / Rmerge(I) obs: 0.08 / Net I/σ(I): 14
Reflection shellResolution: 2.5→2.6 Å / Redundancy: 3.2 % / Rmerge(I) obs: 0.44 / Mean I/σ(I) obs: 3.2 / % possible all: 78

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Processing

Software
NameVersionClassification
REFMAC5.2.0019refinement
XDSdata reduction
XSCALEdata scaling
PHASERphasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: PDB ENTRY 1HG1
Resolution: 2.5→20 Å / Cor.coef. Fo:Fc: 0.955 / Cor.coef. Fo:Fc free: 0.91 / SU B: 23.414 / SU ML: 0.273 / TLS residual ADP flag: LIKELY RESIDUAL / Cross valid method: THROUGHOUT / ESU R: 1.83 / ESU R Free: 0.341 / Stereochemistry target values: MAXIMUM LIKELIHOOD / Details: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS.
RfactorNum. reflection% reflectionSelection details
Rfree0.266 4197 5 %RANDOM
Rwork0.188 ---
obs0.192 79398 96.1 %-
Solvent computationIon probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.4 Å / Solvent model: BABINET MODEL WITH MASK
Displacement parametersBiso mean: 34.63 Å2
Baniso -1Baniso -2Baniso -3
1-0.13 Å20 Å20 Å2
2---1.57 Å20 Å2
3---1.44 Å2
Refinement stepCycle: LAST / Resolution: 2.5→20 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms18719 0 72 708 19499
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.0090.02219064
X-RAY DIFFRACTIONr_bond_other_d
X-RAY DIFFRACTIONr_angle_refined_deg1.2921.98225885
X-RAY DIFFRACTIONr_angle_other_deg
X-RAY DIFFRACTIONr_dihedral_angle_1_deg5.95252510
X-RAY DIFFRACTIONr_dihedral_angle_2_deg38.78824.278699
X-RAY DIFFRACTIONr_dihedral_angle_3_deg18.201153202
X-RAY DIFFRACTIONr_dihedral_angle_4_deg18.21115112
X-RAY DIFFRACTIONr_chiral_restr0.0830.23123
X-RAY DIFFRACTIONr_gen_planes_refined0.0040.0214052
X-RAY DIFFRACTIONr_gen_planes_other
X-RAY DIFFRACTIONr_nbd_refined0.2270.210760
X-RAY DIFFRACTIONr_nbd_other
X-RAY DIFFRACTIONr_nbtor_refined0.3080.213149
X-RAY DIFFRACTIONr_nbtor_other
X-RAY DIFFRACTIONr_xyhbond_nbd_refined0.1570.21233
X-RAY DIFFRACTIONr_xyhbond_nbd_other
X-RAY DIFFRACTIONr_metal_ion_refined
X-RAY DIFFRACTIONr_metal_ion_other
X-RAY DIFFRACTIONr_symmetry_vdw_refined0.1830.227
X-RAY DIFFRACTIONr_symmetry_vdw_other
X-RAY DIFFRACTIONr_symmetry_hbond_refined0.2910.26
X-RAY DIFFRACTIONr_symmetry_hbond_other
X-RAY DIFFRACTIONr_symmetry_metal_ion_refined
X-RAY DIFFRACTIONr_symmetry_metal_ion_other
X-RAY DIFFRACTIONr_mcbond_it0.3331.512754
X-RAY DIFFRACTIONr_mcbond_other
X-RAY DIFFRACTIONr_mcangle_it0.589220194
X-RAY DIFFRACTIONr_mcangle_other
X-RAY DIFFRACTIONr_scbond_it0.97536902
X-RAY DIFFRACTIONr_scbond_other
X-RAY DIFFRACTIONr_scangle_it1.5724.55691
X-RAY DIFFRACTIONr_scangle_other
X-RAY DIFFRACTIONr_long_range_B_refined
X-RAY DIFFRACTIONr_long_range_B_other
X-RAY DIFFRACTIONr_rigid_bond_restr
X-RAY DIFFRACTIONr_sphericity_free
X-RAY DIFFRACTIONr_sphericity_bonded
LS refinement shellResolution: 2.5→2.57 Å / Total num. of bins used: 20 /
RfactorNum. reflection
Rfree0.344 252
Rwork0.252 4598
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)
19.942911.1362-7.280418.7761-6.56679.6750.5258-0.32180.15010.8698-0.16960.1538-0.6606-0.4336-0.35620.27010.1483-0.01880.0233-0.00070.0218-8.29419.32462.54
22.8106-2.35464.0776.1922-0.97767.32270.27590.71220.55910.2928-0.19750.8818-0.99790.4137-0.07840.29920.0229-0.073-0.00470.00730.2266-10.53623.62855.003
30.61190.09260.03091.3861-0.20160.8805-0.0122-0.11310.0580.3216-0.06210.0324-0.2035-0.12530.07430.32380.0146-0.01040.02480.01420.0184-1.71210.80860.797
41.27650.1498-0.52131.74220.082.3682-0.0431-0.18630.1450.3967-0.0498-0.2937-0.21610.43030.09290.2095-0.0587-0.16510.08780.06390.039423.2747.53761.468
51.68230.1928-0.14167.0757-0.28993.08960.10730.13810.20530.01020.02760.6645-0.3663-0.4379-0.13480.23130.2022-0.10510.15350.00010.151-18.49315.67328.809
60.6213-0.0364-0.48932.04081.25073.04430.05620.34760.0019-0.2293-0.17150.1903-0.3593-0.26190.11530.23190.1363-0.10230.14270.01680.0412-10.71711.36925.234
70.695-0.0867-0.13681.321-0.4910.77680.0750.1834-0.1107-0.0034-0.06940.0663-0.0776-0.0249-0.00550.16230.0785-0.0730.0313-0.03410.0211-7.099-3.76328.304
80.78240.3877-0.67662.1675-0.71243.393-0.16660.0485-0.3042-0.26480.14380.18970.0713-0.09480.02290.17520.0493-0.0866-0.0222-0.05030.0569-5.801-15.33521.946
97.05720.9175-0.07171.03442.61437.5229-0.01470.1399-0.47160.0501-0.0492-0.50070.88330.75590.0640.24890.1780.01040.15020.09390.169430.131-12.79330.459
101.06410.0049-0.49381.3836-0.11861.33020.06250.1339-0.0859-0.1541-0.0963-0.19310.04360.34090.03370.15130.0681-0.00180.17360.05410.068722.48-1.2630.817
111.52730.4332-0.77280.2735-0.27512.39830.12830.25880.22690.02380.0418-0.51860.1740.2743-0.17010.2639-0.1215-0.03170.09250.09670.150427.25619.33143.742
121.20030.01320.50192.10860.52842.7280.03470.06840.31230.0722-0.0578-0.219-0.34210.32060.02310.2085-0.0563-0.03130.03630.07940.044319.09422.29140.902
133.72691.20190.56683.5061-1.42373.60660.18180.0076-0.4433-0.1285-0.0882-0.29490.7930.5875-0.09350.32170.17990.03140.05370.00190.271213.676-29.71348.353
141.2813-0.30060.67561.346-0.43890.80040.08270.0137-0.26640.0871-0.10270.05370.2466-0.02040.020.29440.0454-0.0395-0.02670.00980.15932.698-25.61250.436
151.61770.2778-0.39110.8904-0.43541.83380.00890.0644-0.18440.2636-0.0439-0.00490.18020.05760.03510.25450.0467-0.0314-0.02470.04010.07152.899-13.33753.882
161.58160.21161.15190.9836-0.67952.7610.09520.0801-0.27720.1295-0.09960.31820.1512-0.22220.00440.10590.01240.0019-0.0079-0.0620.1892-20.267-14.59343.779
173.9269-1.00531.581.5876-1.01433.36090.18250.2940.1455-0.179-0.2241-0.1681-0.72820.78150.04160.2812-0.15770.00360.23430.0617-0.11945.50421.34-45.484
180.13920.22970.43531.0117-0.38943.30010.04940.288-0.0218-0.0484-0.1184-0.0847-0.00890.34850.06890.163-0.0362-0.05870.2083-0.0366-0.05682.1679.725-37.738
196.7668-0.604-2.69180.1179-0.30035.63570.05080.58550.0533-0.0787-0.16920.2032-0.3958-0.39390.11840.2072-0.0168-0.11190.0356-0.07860.0287-19.4913.3-43.35
201.2288-0.59170.32771.06580.12112.91340.1250.11120.1225-0.1951-0.07940.1649-0.1016-0.211-0.04560.17690.0218-0.11960.1333-0.05150.0173-24.0147.256-41.681
215.774-2.317-0.89036.7443-0.37734.46710.1652-0.33930.25080.5158-0.188-0.3364-1.20290.73030.02290.3158-0.4627-0.17240.3456-0.0103-0.055615.3318.386-7.258
2210.9725-1.26676.92591.7189-2.24935.70811.78050.99711.043-0.7859-0.4715-2.3935-0.79443.1019-1.3090.7122-0.1545-0.14530.6885-0.02830.480819.6121.572-15.926
233.4112-0.1243-0.76511.81970.17811.37510.1914-0.72220.10280.2743-0.1889-0.3085-0.52530.6355-0.00250.2915-0.3667-0.21250.54160.0856-0.137616.2711.041-2.735
241.1967-0.00210.13841.885-0.74211.62980.0572-0.334-0.26690.287-0.2022-0.1228-0.1970.31030.14510.0814-0.1089-0.08960.24350.0832-0.00736.666-2.734-6.637
251.6512-0.7513-0.29934.1849-0.69331.7144-0.1351-0.1754-0.08690.49510.22060.4588-0.0189-0.4437-0.08550.0488-0.0548-0.00910.32010.0484-0.0531-32.65-4.147-9.205
260.41450.3599-0.05231.5432-0.97941.75450.0516-0.1285-0.12330.1422-0.00770.1859-0.2122-0.2559-0.04390.1293-0.0063-0.02180.1489-0.032-0.0092-21.2114.55-15.059
274.854-3.06250.30532.4796-1.58373.55450.0229-0.23230.2094-0.02520.03110.4781-0.61320.2034-0.0540.3533-0.028-0.04780.0037-0.0608-0.0044-17.51525.19-26.435
284.191-2.55932.29078.4648-1.27292.37160.0128-0.35630.26360.4223-0.04460.1256-0.4927-0.14440.03180.33190.04480.02190.0644-0.05-0.0932-20.77621.748-15.012
294.5034-0.64160.57676.9169-2.02651.36510.4094-0.1584-0.8416-0.0352-0.16060.30560.6353-0.2825-0.24880.0895-0.0885-0.0862-0.05-0.0910.2896-11.869-29.292-30.851
301.4649-0.29920.39531.2521-0.01950.73150.10590.1044-0.2242-0.2403-0.0788-0.04890.21210.024-0.02710.2181-0.0291-0.06470.0633-0.05660.1033-3.927-18.04-32.403
312.2173-0.21870.12281.58780.31741.58960.09710.1034-0.4574-0.1131-0.241-0.25060.07920.59850.14390.05270.0231-0.06380.35370.11180.246219.866-13.917-22.975
327.4417-2.48551.13810.95170.3834.96620.19790.18-0.3234-0.2191-0.342-0.19630.50090.70390.1440.16680.11060.00160.4021-0.04170.114817.919-13.55-32.027
Refinement TLS group
IDRefine-IDRefine TLS-IDAuth asym-IDAuth seq-ID
1X-RAY DIFFRACTION1A3 - 20
2X-RAY DIFFRACTION2A21 - 50
3X-RAY DIFFRACTION3A51 - 205
4X-RAY DIFFRACTION4A206 - 327
5X-RAY DIFFRACTION5B3 - 70
6X-RAY DIFFRACTION6B71 - 153
7X-RAY DIFFRACTION7B154 - 256
8X-RAY DIFFRACTION8B257 - 327
9X-RAY DIFFRACTION9C3 - 50
10X-RAY DIFFRACTION10C51 - 205
11X-RAY DIFFRACTION11C206 - 240
12X-RAY DIFFRACTION12C241 - 327
13X-RAY DIFFRACTION13D3 - 55
14X-RAY DIFFRACTION14D56 - 143
15X-RAY DIFFRACTION15D144 - 211
16X-RAY DIFFRACTION16D212 - 327
17X-RAY DIFFRACTION17E3 - 91
18X-RAY DIFFRACTION18E92 - 202
19X-RAY DIFFRACTION19E203 - 234
20X-RAY DIFFRACTION20E235 - 327
21X-RAY DIFFRACTION21F3 - 19
22X-RAY DIFFRACTION22F20 - 47
23X-RAY DIFFRACTION23F48 - 100
24X-RAY DIFFRACTION24F101 - 327
25X-RAY DIFFRACTION25G3 - 87
26X-RAY DIFFRACTION26G88 - 242
27X-RAY DIFFRACTION27G243 - 289
28X-RAY DIFFRACTION28G290 - 327
29X-RAY DIFFRACTION29H3 - 62
30X-RAY DIFFRACTION30H63 - 201
31X-RAY DIFFRACTION31H202 - 289
32X-RAY DIFFRACTION32H290 - 327

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