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- PDB-4kgv: The R state structure of E. coli ATCase with ATP bound -

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Basic information

Entry
Database: PDB / ID: 4kgv
TitleThe R state structure of E. coli ATCase with ATP 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
ADENOSINE-5'-TRIPHOSPHATE / N-(PHOSPHONACETYL)-L-ASPARTIC ACID / : / :
Similarity search - Component
Biological speciesEscherichia coli (E. coli)
MethodX-RAY DIFFRACTION / MOLECULAR REPLACEMENT / Resolution: 2.1 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 1.1Sep 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

Downloads & links

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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)104,61710
Polymers102,9614
Non-polymers1,6556
Water12,394688
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)313,85130
Polymers308,88412
Non-polymers4,96618
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 area30910 Å2
ΔGint-118 kcal/mol
Surface area103140 Å2
MethodPISA
Unit cell
Length a, b, c (Å)121.070, 121.070, 155.490
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-700-

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, 694 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-ATP / ADENOSINE-5'-TRIPHOSPHATE / Adenosine triphosphate


Mass: 507.181 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: C10H16N5O13P3 / Comment: ATP, energy-carrying molecule*YM
#6: Water ChemComp-HOH / water / Water


Mass: 18.015 Da / Num. of mol.: 688 / 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: 3.2 Å3/Da / Density % sol: 61.5 %
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: ROTATING ANODE / Type: RIGAKU MICROMAX-007 HF / Wavelength: 1.542 Å
DetectorType: RIGAKU RAXIS IV++ / Detector: IMAGE PLATE / Date: Aug 9, 2012 / Details: Osmic VariMax Optics
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 1.542 Å / Relative weight: 1
ReflectionResolution: 2.1→38.4 Å / Num. all: 77283 / Num. obs: 77283 / % possible obs: 100 % / Observed criterion σ(I): -3 / Redundancy: 5.34 % / Rmerge(I) obs: 0.054 / Net I/σ(I): 11.6
Reflection shell
Resolution (Å)Redundancy (%)Rmerge(I) obsMean I/σ(I) obsDiffraction-ID% possible all
2.1-2.185.30.423.11100
2.18-2.265.260.3523.71100
2.26-2.375.340.2844.31100
2.37-2.495.350.2165.41100
2.49-2.655.360.1636.91100
2.65-2.855.360.1268.51100
2.85-3.145.40.09111.51100
3.14-3.595.410.06615.1199.9
3.59-4.525.370.05121.5199.9
4.52-38.45.250.03334.4199.8

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Processing

Software
NameVersionClassificationNB
d*TREKdata scaling
d*TREK9.9.8.8Ddata reduction
PHENIX1.8.1_1168refinement
PDB_EXTRACT3.11data extraction
CrystalCleardata collection
PHENIXphasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: PDB entry 1D09
Resolution: 2.1→38.4 Å / Occupancy max: 1 / Occupancy min: 1 / SU ML: 0.21 / σ(F): 0 / Phase error: 22.74 / Stereochemistry target values: ML
RfactorNum. reflection% reflection
Rfree0.2075 3880 5.02 %
Rwork0.1675 --
obs0.1695 77271 99.94 %
all-77271 -
Solvent computationShrinkage radii: 0.9 Å / VDW probe radii: 1.11 Å / Solvent model: FLAT BULK SOLVENT MODEL
Displacement parametersBiso mean: 48.3145 Å2
Refinement stepCycle: LAST / Resolution: 2.1→38.4 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms7098 0 96 688 7882
Refine LS restraints
Refine-IDTypeDev idealNumber
X-RAY DIFFRACTIONf_bond_d0.0077320
X-RAY DIFFRACTIONf_angle_d1.0679944
X-RAY DIFFRACTIONf_dihedral_angle_d14.692740
X-RAY DIFFRACTIONf_chiral_restr0.0721150
X-RAY DIFFRACTIONf_plane_restr0.0051284
LS refinement shell
Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkRefine-ID% reflection obs (%)
2.1-2.12560.26761340.22572609X-RAY DIFFRACTION100
2.1256-2.15250.29431340.21442566X-RAY DIFFRACTION100
2.1525-2.18080.23461200.21232638X-RAY DIFFRACTION100
2.1808-2.21070.25551210.20942601X-RAY DIFFRACTION100
2.2107-2.24230.23891360.20822591X-RAY DIFFRACTION100
2.2423-2.27570.23511260.20982564X-RAY DIFFRACTION100
2.2757-2.31130.24421460.21492611X-RAY DIFFRACTION100
2.3113-2.34920.25771440.19892606X-RAY DIFFRACTION100
2.3492-2.38970.2481400.20152589X-RAY DIFFRACTION100
2.3897-2.43310.22881210.19862616X-RAY DIFFRACTION100
2.4331-2.47990.26711410.19742594X-RAY DIFFRACTION100
2.4799-2.53050.24371390.19612591X-RAY DIFFRACTION100
2.5305-2.58550.22261430.19332627X-RAY DIFFRACTION100
2.5855-2.64570.28551480.18932565X-RAY DIFFRACTION100
2.6457-2.71180.25571350.19412603X-RAY DIFFRACTION100
2.7118-2.78510.22971420.18042601X-RAY DIFFRACTION100
2.7851-2.8670.24921310.18342640X-RAY DIFFRACTION100
2.867-2.95960.23191380.18222614X-RAY DIFFRACTION100
2.9596-3.06530.25131460.18122593X-RAY DIFFRACTION100
3.0653-3.1880.22081270.18292665X-RAY DIFFRACTION100
3.188-3.3330.19311490.17992586X-RAY DIFFRACTION100
3.333-3.50860.21271400.16632643X-RAY DIFFRACTION100
3.5086-3.72820.18911430.15332629X-RAY DIFFRACTION100
3.7282-4.01580.19821360.14252641X-RAY DIFFRACTION100
4.0158-4.41940.16951510.13272654X-RAY DIFFRACTION100
4.4194-5.05770.17191600.12892649X-RAY DIFFRACTION100
5.0577-6.36740.1891440.1732713X-RAY DIFFRACTION100
6.3674-38.40820.18191450.16052792X-RAY DIFFRACTION99
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)
1-0.0045-0.02230.00480.16280.03030.032-0.01280.19780.1832-0.122-0.020.203-0.4189-0.0133-0.00010.27960.0504-0.07260.35880.10080.370335.45843.95643.1561
2-0.0391-0.0150.02590.24070.16450.16240.12890.624-0.1-0.76450.14520.438-0.1596-0.65730.01430.37490.0287-0.13720.46610.01210.370534.085232.5295-2.9492
31.30220.6016-0.15131.32140.3890.2990.06570.03510.0528-0.14030.04470.031-0.0611-0.015600.2120.03250.00180.2330.03820.220649.319841.519610.1394
40.3992-0.1868-0.3951.20110.08540.8583-0.0153-0.0495-0.1579-0.03040.12310.5019-0.1814-0.12340.04180.1704-0.0195-0.03080.32440.11660.423728.331528.513513.6206
50.6370.40230.1520.39740.41580.81620.0758-0.7044-0.4963-0.1237-0.29180.8053-0.1896-1.3166-0.39230.1108-0.14060.1340.45980.31340.842820.66827.476518.9394
60.33110.24680.03880.6225-0.42740.53920.1732-0.3178-0.15540.60890.10420.4882-0.160.09240.06250.3236-0.01290.04180.47590.14170.367436.927228.67727.3792
70.8582-0.01440.02091.70670.85080.3441-0.3814-0.3724-0.65511.21070.37041.00240.5758-0.1644-0.00590.2218-0.13180.04180.40180.22670.811730.208313.569820.7756
80.1525-0.17840.17680.0238-0.11850.15370.1409-0.2986-0.49480.0081-0.00150.41940.1174-0.29320.00310.2944-0.0275-0.04920.29150.15220.636837.841715.752816.2824
90.6369-0.2329-0.00250.05260.04160.0820.04020.3232-0.109-0.43060.03770.13230.135-0.0868-00.23660.0074-0.06350.32240.02640.26441.541331.7350.0708
100.47260.0171-0.1960.89630.35940.84760.0180.25030.01670.0616-0.21260.0784-0.014-0.2058-0.00180.44520.2057-0.02480.5215-0.14240.430619.909974.863331.3316
110.5538-0.8098-0.7731.25381.25531.422-0.1038-0.1525-0.0287-0.0697-0.2480.1327-0.0036-0.5501-0.15330.21620.1149-0.0260.5387-0.10850.456124.939859.053723.3066
120.92050.60720.13730.81520.22990.53920.1001-0.26350.11040.08520.01580.0789-0.0793-0.091700.35350.03630.03280.363-0.03580.274659.659654.572778.5376
130.75290.60930.69560.8024-0.06510.3905-0.04160.03310.035-0.04150.03380.0605-0.0799-0.013600.26920.01570.02330.27530.00830.187953.74947.232467.4281
140.89660.0312-0.71970.1699-0.19280.60290.05980.04310.2078-0.04260.0662-0.0001-0.38080.064200.4174-0.03510.04820.28280.01530.309670.236166.351868.0435
150.13710.08970.10740.2040.21050.1894-0.28780.40950.5824-0.9030.27160.3651-0.6380.161-0.00170.62420.03240.02590.3090.10860.51467.670173.047558.1076
160.1877-0.2803-0.26510.27740.19330.3875-0.05570.5980.414-0.17440.2066-0.0051-0.07140.39450.00110.4658-0.10320.08050.41760.05620.376974.824162.676758.3436
170.3330.0554-0.36891.2287-0.48310.4993-0.04810.42810.0556-0.30420.1982-0.2271-0.24380.1148-0.01970.4516-0.09920.19390.551-0.02520.375480.493861.93155.3553
180.7709-0.07170.66450.88830.11850.44880.0884-0.12170.10820.0325-0.0249-0.0887-0.07940.040.00010.3089-0.01630.05950.288-0.03570.268273.175254.513572.0204
191.4091-0.12690.67281.18770.90181.2621-0.3363-0.264-0.06130.61630.0394-0.1947-0.11120.0035-0.02340.75980.2225-0.0260.4704-0.08370.387724.841479.401348.0063
201.3522-0.33320.42130.2269-0.40731.3047-0.54510.05280.3023-0.1632-0.0197-0.0391-0.68120.0644-0.47570.66880.0743-0.24750.36320.06720.431445.317568.343955.3827
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 134 )
4X-RAY DIFFRACTION4chain 'A' and (resid 135 through 195 )
5X-RAY DIFFRACTION5chain 'A' and (resid 196 through 223 )
6X-RAY DIFFRACTION6chain 'A' and (resid 224 through 242 )
7X-RAY DIFFRACTION7chain 'A' and (resid 243 through 261 )
8X-RAY DIFFRACTION8chain 'A' and (resid 262 through 284 )
9X-RAY DIFFRACTION9chain 'A' and (resid 285 through 310 )
10X-RAY DIFFRACTION10chain 'B' and (resid 9 through 67 )
11X-RAY DIFFRACTION11chain 'B' and (resid 68 through 153 )
12X-RAY DIFFRACTION12chain 'C' and (resid 1 through 53 )
13X-RAY DIFFRACTION13chain 'C' and (resid 54 through 134 )
14X-RAY DIFFRACTION14chain 'C' and (resid 135 through 187 )
15X-RAY DIFFRACTION15chain 'C' and (resid 188 through 215 )
16X-RAY DIFFRACTION16chain 'C' and (resid 216 through 236 )
17X-RAY DIFFRACTION17chain 'C' and (resid 237 through 261 )
18X-RAY DIFFRACTION18chain 'C' and (resid 262 through 310 )
19X-RAY DIFFRACTION19chain 'D' and (resid 9 through 95 )
20X-RAY DIFFRACTION20chain 'D' and (resid 96 through 153 )

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