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- PDB-4kgx: The R state structure of E. coli ATCase with CTP bound -

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Basic information

Entry
Database: PDB / ID: 4kgx
TitleThe R state structure of E. coli ATCase with CTP bound
Components
  • Aspartate carbamoyltransferase regulatory chain
  • Aspartate carbamoyltransferase
KeywordsTRANSFERASE / pyrimidine nucleotide biosynthesis / feedback inhibition / competing pathway product activation / allostery
Function / homology
Function and homology information


Aspartate carbamoyltransferase regulatory subunit, N-terminal domain / Aspartate carbamoyltransferase regulatory subunit, C-terminal domain / Aspartate/ornithine carbamoyltransferase / SH3 type barrels. / Roll / Alpha-Beta Plaits / Rossmann fold / 2-Layer Sandwich / 3-Layer(aba) Sandwich / Mainly Beta / Alpha Beta
Similarity search - Domain/homology
CYTIDINE-5'-TRIPHOSPHATE / N-(PHOSPHONACETYL)-L-ASPARTIC ACID / : / :
Similarity search - Component
Biological speciesEscherichia coli (E. coli)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 2.202 Å
AuthorsCockrell, G.M. / Zheng, Y. / Guo, W. / Peterson, A.W. / Kantrowitz, E.R.
CitationJournal: Biochemistry / Year: 2013
Title: New Paradigm for Allosteric Regulation of Escherichia coli Aspartate Transcarbamoylase.
Authors: Cockrell, G.M. / Zheng, Y. / Guo, W. / Peterson, A.W. / Truong, J.K. / Kantrowitz, E.R.
History
DepositionApr 29, 2013Deposition site: RCSB / Processing site: RCSB
Revision 1.0Nov 27, 2013Provider: repository / Type: Initial release
Revision 1.1Nov 15, 2017Group: Refinement description / Category: software / Item: _software.classification / _software.name
Revision 1.2Sep 20, 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 / pdbx_struct_conn_angle / struct_conn / struct_site
Item: _database_2.pdbx_DOI / _database_2.pdbx_database_accession ..._database_2.pdbx_DOI / _database_2.pdbx_database_accession / _pdbx_struct_conn_angle.ptnr1_auth_seq_id / _pdbx_struct_conn_angle.ptnr1_label_seq_id / _pdbx_struct_conn_angle.ptnr3_auth_seq_id / _pdbx_struct_conn_angle.ptnr3_label_seq_id / _pdbx_struct_conn_angle.value / _struct_conn.pdbx_dist_value / _struct_conn.ptnr1_auth_asym_id / _struct_conn.ptnr1_auth_seq_id / _struct_conn.ptnr1_label_asym_id / _struct_conn.ptnr1_label_seq_id / _struct_conn.ptnr2_auth_asym_id / _struct_conn.ptnr2_label_asym_id / _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: Aspartate carbamoyltransferase
B: Aspartate carbamoyltransferase regulatory chain
C: Aspartate carbamoyltransferase
D: Aspartate carbamoyltransferase regulatory chain
hetero molecules


Theoretical massNumber of molelcules
Total (without water)104,56910
Polymers102,9614
Non-polymers1,6076
Water12,304683
1
A: Aspartate carbamoyltransferase
B: Aspartate carbamoyltransferase regulatory chain
C: Aspartate carbamoyltransferase
D: Aspartate carbamoyltransferase regulatory chain
hetero molecules

A: Aspartate carbamoyltransferase
B: Aspartate carbamoyltransferase regulatory chain
C: Aspartate carbamoyltransferase
D: Aspartate carbamoyltransferase regulatory chain
hetero molecules

A: Aspartate carbamoyltransferase
B: Aspartate carbamoyltransferase regulatory chain
C: Aspartate carbamoyltransferase
D: Aspartate carbamoyltransferase regulatory chain
hetero molecules


Theoretical massNumber of molelcules
Total (without water)313,70630
Polymers308,88412
Non-polymers4,82218
Water21612
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
crystal symmetry operation2_655-y+1,x-y,z1
crystal symmetry operation3_665-x+y+1,-x+1,z1
Buried area34430 Å2
ΔGint-103 kcal/mol
Surface area100680 Å2
MethodPISA
Unit cell
Length a, b, c (Å)121.237, 121.237, 154.772
Angle α, β, γ (deg.)90.00, 90.00, 120.00
Int Tables number150
Space group name H-MP321
Components on special symmetry positions
IDModelComponents
11A-702-

HOH

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Components

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Protein , 2 types, 4 molecules ACBD

#1: Protein Aspartate carbamoyltransferase / / Aspartate transcarbamylase


Mass: 34337.105 Da / Num. of mol.: 2
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Escherichia coli (E. coli) / Strain: ATCC 55124 / KO11 / Gene: pyrB, EKO11_4066, KO11_22860 / Production host: Escherichia coli (E. coli) / References: UniProt: E8Y328, aspartate carbamoyltransferase
#2: Protein Aspartate carbamoyltransferase regulatory chain


Mass: 17143.625 Da / Num. of mol.: 2
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Escherichia coli (E. coli) / Strain: ATCC 55124 / KO11 / Gene: pyrI, EKO11_4067, KO11_22855 / Production host: Escherichia coli (E. coli) / References: UniProt: E8Y329, aspartate carbamoyltransferase

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Non-polymers , 4 types, 689 molecules

#3: Chemical ChemComp-PAL / N-(PHOSPHONACETYL)-L-ASPARTIC ACID


Mass: 255.119 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: C6H10NO8P
#4: Chemical ChemComp-ZN / ZINC ION


Mass: 65.409 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: Zn
#5: Chemical ChemComp-CTP / CYTIDINE-5'-TRIPHOSPHATE / Cytidine triphosphate


Mass: 483.156 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: C9H16N3O14P3
#6: Water ChemComp-HOH / water / Water


Mass: 18.015 Da / Num. of mol.: 683 / 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: 3.19 Å3/Da / Density % sol: 61.43 %
Crystal growTemperature: 293 K / Method: microdialysis / pH: 5.9
Details: 50 mM maleic acid, 1 mM PALA, 3 mM sodium azide, pH 5.9, Microdialysis, temperature 293K

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: NSLS / Beamline: X29A / Wavelength: 1.075 Å
DetectorType: ADSC QUANTUM 315 / Detector: CCD / Date: Oct 19, 2011
RadiationMonochromator: Rosenbaum-Rock double crystal sagittal focusing monochromator
Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 1.075 Å / Relative weight: 1
ReflectionResolution: 2.2→50 Å / Num. all: 67084 / Num. obs: 67084 / % possible obs: 100 % / Observed criterion σ(I): -3 / Redundancy: 22.4 % / Rmerge(I) obs: 0.075 / Net I/σ(I): 10.8
Reflection shell
Resolution (Å)Redundancy (%)Rmerge(I) obsDiffraction-ID% possible all
2.2-2.2818.90.6471100
2.28-2.3719.20.531100
2.37-2.4819.50.3831100
2.48-2.61200.2711100
2.61-2.7720.70.1981100
2.77-2.9921.50.1421100
2.99-3.29220.0971100
3.29-3.76220.0741100
3.76-4.7428.20.0641100
4.74-5031.90.0431100

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Processing

Software
NameVersionClassificationNB
DENZOdata reduction
SCALEPACKdata scaling
PHENIX1.8.1_1168refinement
PDB_EXTRACT3.11data extraction
HKL-2000data collection
HKL-2000data reduction
HKL-2000data scaling
PHENIXphasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: PDB entry 1D09
Resolution: 2.202→49.715 Å / Occupancy max: 1 / Occupancy min: 1 / SU ML: 0.19 / σ(F): 0 / Phase error: 19.3 / Stereochemistry target values: ML
RfactorNum. reflection% reflection
Rfree0.1924 3395 5.06 %
Rwork0.1559 --
obs0.1578 67071 99.96 %
all-67071 -
Solvent computationShrinkage radii: 1.3 Å / VDW probe radii: 1.4 Å / Solvent model: FLAT BULK SOLVENT MODEL
Displacement parametersBiso mean: 39.2568 Å2
Refinement stepCycle: LAST / Resolution: 2.202→49.715 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms7082 0 92 683 7857
Refine LS restraints
Refine-IDTypeDev idealNumber
X-RAY DIFFRACTIONf_bond_d0.0077298
X-RAY DIFFRACTIONf_angle_d1.0659912
X-RAY DIFFRACTIONf_dihedral_angle_d14.2992736
X-RAY DIFFRACTIONf_chiral_restr0.0721147
X-RAY DIFFRACTIONf_plane_restr0.0041283
LS refinement shell
Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkRefine-ID% reflection obs (%)
2.202-2.23320.26491290.20732617X-RAY DIFFRACTION100
2.2332-2.26660.21641450.19932604X-RAY DIFFRACTION100
2.2666-2.3020.24551480.19772610X-RAY DIFFRACTION100
2.302-2.33970.23631430.19212666X-RAY DIFFRACTION100
2.3397-2.38010.25071380.18052605X-RAY DIFFRACTION100
2.3801-2.42330.21321520.17612631X-RAY DIFFRACTION100
2.4233-2.46990.21681240.16882620X-RAY DIFFRACTION100
2.4699-2.52040.20781360.15972638X-RAY DIFFRACTION100
2.5204-2.57520.21111340.16282633X-RAY DIFFRACTION100
2.5752-2.63510.21691560.16432630X-RAY DIFFRACTION100
2.6351-2.7010.22811530.16282604X-RAY DIFFRACTION100
2.701-2.7740.21571410.15912635X-RAY DIFFRACTION100
2.774-2.85560.24111240.15982635X-RAY DIFFRACTION100
2.8556-2.94780.21791370.16652647X-RAY DIFFRACTION100
2.9478-3.05310.21961440.16622652X-RAY DIFFRACTION100
3.0531-3.17530.19211690.16512621X-RAY DIFFRACTION100
3.1753-3.31980.24221330.17182647X-RAY DIFFRACTION100
3.3198-3.49480.18351640.15462650X-RAY DIFFRACTION100
3.4948-3.71370.15731280.14762668X-RAY DIFFRACTION100
3.7137-4.00030.16051410.14382682X-RAY DIFFRACTION100
4.0003-4.40270.18291440.13062679X-RAY DIFFRACTION100
4.4027-5.03920.17711320.12822714X-RAY DIFFRACTION100
5.0392-6.34680.16661450.16762730X-RAY DIFFRACTION100
6.3468-49.72780.16771350.15032858X-RAY DIFFRACTION100
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)
10.0047-0.0070.00290.0058-0.00460.0145-0.00860.02720.0099-0.0249-0.03280.0092-0.02220.0154-0.03230.10780.1106-0.09330.16070.15820.28735.595144.16893.1506
20.0116-0.001-0.00270.004-0.00230.0048-0.01020.0278-0.0076-0.01130.01230.0099-0.0173-0.01240.0090.0770.0317-0.15390.1723-0.01360.174434.207932.7382-2.9601
30.0124-0.0119-0.00750.00930.0090.0041-0.01490.08930.0452-0.06560.0616-0.0171-0.0316-0.00040.01930.14940.00330.00810.16580.0350.120350.66736.80092.4508
40.1529-0.0413-0.03860.10810.05590.17020.0667-0.05260.0909-0.03550.02250.0361-0.02420.00830.18310.09320.0260.02110.11580.02030.145849.118442.963512.2376
50.03810.03610.00420.11350.03070.04260.0262-0.0151-0.1623-0.01850.09250.26590.0188-0.05050.1233-0.0368-0.0286-0.0180.20780.10790.410728.470628.537211.7876
60.00960.00780.00320.00830.010.023-0.0154-0.01850.02820.0126-0.01690.07040.0049-0.0183-0.03610.06410.05330.13550.32420.14190.351323.704235.210422.4294
70.02340.0109-0.02960.0031-0.01310.0353-0.0132-0.0128-0.05060.0167-0.02910.0350.0601-0.0476-0.03540.0356-0.08330.07260.19770.23560.609322.686922.084918.3871
80.01470.0141-0.0120.0111-0.01240.01010.0277-0.0622-0.05750.0805-0.00510.03430.01430.01410.04650.19840.02630.13110.35720.21710.138637.142626.48229.691
90.0221-0.0365-0.00050.0586-0.001-0.0006-0.0228-0.0267-0.09270.02580.01230.06150.0369-0.0197-0.02860.0633-0.08840.03830.04370.22590.565134.474814.287515.882
100.0266-0.0185-0.0320.08530.03320.04540.01880.085-0.0834-0.10720.02740.0357-0.0391-0.02390.02580.04670.0105-0.09020.11890.03730.172241.660931.97710.0499
110.0055-0.0050.00760.0051-0.00760.0134-0.01730.02870.0220.0086-0.04110.0187-0.0064-0.027-0.02140.29480.1912-0.01660.2989-0.07630.271516.833476.793329.6537
120.02310.0096-0.01870.0168-0.00540.023-0.06690.0156-0.04940.048-0.04740.0450.03380.0044-0.05950.32830.3572-0.01050.4462-0.22610.242421.053874.223931.7775
130.04230.0098-0.04550.0045-0.01090.0488-0.07460.0443-0.0706-0.0525-0.0394-0.0049-0.0406-0.0754-0.04730.19250.2926-0.0950.4849-0.22880.373818.369269.847820.985
140.0070.0057-0.01040.0232-0.0110.0129-0.0484-0.12010.01970.0712-0.04220.16270.0534-0.0938-0.02340.07560.03380.07320.3188-0.07430.342830.304751.230324.8735
150.0189-0.00610.010.02250.00710.01450.0744-0.17040.1210.08920.03730.0567-0.0699-0.0410.11820.21690.0340.02960.2675-0.01440.090859.683654.611678.2372
160.0319-0.0028-0.04250.07120.01610.10450.03490.05210.0853-0.07920.00370.0335-0.1480.01520.13520.20770.01720.03270.20250.0290.094259.047752.315267.702
170.0498-0.00810.00590.0214-0.01950.03670.01910.29130.2217-0.25390.08060.0144-0.1161-0.01560.030.4165-0.05030.09750.25170.06570.233569.862567.893359.0367
180.00490.003-0.00780.0002-0.0030.01460.00410.03960.013-0.0498-0.028-0.03840.00190.0005-0.0040.3361-0.1260.22550.3999-0.09580.263684.768763.173158.8509
190.03870.00960.01890.0699-0.02020.0155-0.0053-0.06880.02050.061-0.0217-0.0745-0.0317-0.0571-0.03670.2496-0.02630.06810.2635-0.02030.161773.209254.593271.7426
200.0626-0.0935-0.0650.17410.12150.0862-0.1539-0.0894-0.02570.2787-0.00180.06810.08040.0233-0.13320.60520.2822-0.05240.3529-0.1070.189326.189778.758947.0727
210.00160.00270.00090.00890.00580.02240.0075-0.03960.00380.0568-0.06010.0137-0.0134-0.00880.00010.78750.3294-0.15660.4892-0.17380.424628.836681.220657.208
220.03060.0008-0.02310.03770.00860.0257-0.0661-0.01880.0365-0.0223-0.02180.03960.00710.0339-0.05570.37590.0639-0.18130.25290.08760.183845.958165.857654.7534
230.00090.0041-0.00260.0157-0.00630.002-0.06010.03790.0262-0.0406-0.0158-0.0068-0.01810.0224-0.0450.4593-0.0033-0.15190.23690.16720.271348.366768.195753.7608
Refinement TLS group
IDRefine-IDRefine TLS-IDSelection details
1X-RAY DIFFRACTION1chain 'A' and (resid 1 through 16 )
2X-RAY DIFFRACTION2chain 'A' and (resid 17 through 32 )
3X-RAY DIFFRACTION3chain 'A' and (resid 33 through 53 )
4X-RAY DIFFRACTION4chain 'A' and (resid 54 through 134 )
5X-RAY DIFFRACTION5chain 'A' and (resid 135 through 187 )
6X-RAY DIFFRACTION6chain 'A' and (resid 188 through 204 )
7X-RAY DIFFRACTION7chain 'A' and (resid 205 through 227 )
8X-RAY DIFFRACTION8chain 'A' and (resid 228 through 250 )
9X-RAY DIFFRACTION9chain 'A' and (resid 251 through 284 )
10X-RAY DIFFRACTION10chain 'A' and (resid 285 through 310 )
11X-RAY DIFFRACTION11chain 'B' and (resid 9 through 25 )
12X-RAY DIFFRACTION12chain 'B' and (resid 26 through 67 )
13X-RAY DIFFRACTION13chain 'B' and (resid 68 through 104 )
14X-RAY DIFFRACTION14chain 'B' and (resid 105 through 153 )
15X-RAY DIFFRACTION15chain 'C' and (resid 1 through 53 )
16X-RAY DIFFRACTION16chain 'C' and (resid 54 through 166 )
17X-RAY DIFFRACTION17chain 'C' and (resid 167 through 242 )
18X-RAY DIFFRACTION18chain 'C' and (resid 243 through 261 )
19X-RAY DIFFRACTION19chain 'C' and (resid 262 through 310 )
20X-RAY DIFFRACTION20chain 'D' and (resid 11 through 87 )
21X-RAY DIFFRACTION21chain 'D' and (resid 88 through 104 )
22X-RAY DIFFRACTION22chain 'D' and (resid 105 through 134 )
23X-RAY DIFFRACTION23chain 'D' and (resid 135 through 153 )

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