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- PDB-4zk7: Crystal structure of rescued two-component self-assembling tetrah... -

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Basic information

Entry
Database: PDB / ID: 4zk7
TitleCrystal structure of rescued two-component self-assembling tetrahedral cage T33-31
Components
  • Chorismate mutase
  • Divalent-cation tolerance protein CutA
KeywordsPROTEIN BINDING / Tetrahedral / computational design / Rosetta / self-assembly / symmetric / nanomaterial / solubility
Function / homology
Function and homology information


chorismate metabolic process / chorismate mutase / chorismate mutase activity / response to metal ion / aromatic amino acid family biosynthetic process / amino acid biosynthetic process / cytoplasm
Similarity search - Function
Chorismate mutase, AroH class / Chorismate mutase type I / Chorismate mutase domain profile. / Divalent ion tolerance protein, CutA / CutA1 divalent ion tolerance protein / RutC-like / RutC-like superfamily / Alpha-Beta Plaits - #120 / Nitrogen regulatory PII-like, alpha/beta / Nitrogen regulatory protein PII/ATP phosphoribosyltransferase, C-terminal ...Chorismate mutase, AroH class / Chorismate mutase type I / Chorismate mutase domain profile. / Divalent ion tolerance protein, CutA / CutA1 divalent ion tolerance protein / RutC-like / RutC-like superfamily / Alpha-Beta Plaits - #120 / Nitrogen regulatory PII-like, alpha/beta / Nitrogen regulatory protein PII/ATP phosphoribosyltransferase, C-terminal / 60s Ribosomal Protein L30; Chain: A; / Alpha-Beta Plaits / 2-Layer Sandwich / Alpha Beta
Similarity search - Domain/homology
Chorismate mutase AroH / Divalent-cation tolerance protein CutA
Similarity search - Component
Biological speciesThermus thermophilus (bacteria)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / molecular replacement / Resolution: 3.4 Å
AuthorsLiu, Y. / Cascio, D. / Sawaya, M.R. / Bale, J. / Collazo, M.J. / Park, R. / King, N. / Baker, D. / Yeates, T.
CitationJournal: Protein Sci. / Year: 2015
Title: Structure of a designed tetrahedral protein assembly variant engineered to have improved soluble expression.
Authors: Bale, J.B. / Park, R.U. / Liu, Y. / Gonen, S. / Gonen, T. / Cascio, D. / King, N.P. / Yeates, T.O. / Baker, D.
History
DepositionApr 30, 2015Deposition site: RCSB / Processing site: RCSB
Revision 1.0Jul 29, 2015Provider: repository / Type: Initial release
Revision 1.1Oct 7, 2015Group: Database references
Revision 1.2Nov 1, 2017Group: Author supporting evidence / Database references / Derived calculations
Category: citation / pdbx_struct_assembly_auth_evidence / pdbx_struct_oper_list
Item: _citation.journal_id_CSD / _pdbx_struct_oper_list.symmetry_operation
Revision 1.3Mar 6, 2024Group: Data collection / Database references / Category: chem_comp_atom / chem_comp_bond / database_2
Item: _database_2.pdbx_DOI / _database_2.pdbx_database_accession

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: Chorismate mutase
B: Chorismate mutase
C: Chorismate mutase
D: Chorismate mutase
E: Chorismate mutase
F: Chorismate mutase
G: Chorismate mutase
H: Chorismate mutase
I: Chorismate mutase
J: Chorismate mutase
K: Chorismate mutase
L: Chorismate mutase
M: Divalent-cation tolerance protein CutA
N: Divalent-cation tolerance protein CutA
O: Divalent-cation tolerance protein CutA
P: Divalent-cation tolerance protein CutA
Q: Divalent-cation tolerance protein CutA
R: Divalent-cation tolerance protein CutA
S: Divalent-cation tolerance protein CutA
T: Divalent-cation tolerance protein CutA
U: Divalent-cation tolerance protein CutA
V: Divalent-cation tolerance protein CutA
W: Divalent-cation tolerance protein CutA
X: Divalent-cation tolerance protein CutA


Theoretical massNumber of molelcules
Total (without water)324,48124
Polymers324,48124
Non-polymers00
Water0
1


  • Idetical with deposited unit
  • defined by author&software
  • Evidence: gel filtration, scanning transmission electron microscopy
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area68660 Å2
ΔGint-374 kcal/mol
Surface area87520 Å2
MethodPISA
Unit cell
Length a, b, c (Å)121.100, 128.400, 204.700
Angle α, β, γ (deg.)90.000, 90.000, 90.000
Int Tables number19
Space group name H-MP212121
DetailsTetrahedral cage from two protein components by gel filtration, SEC, EM

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Components

#1: Protein
Chorismate mutase /


Mass: 14495.646 Da / Num. of mol.: 12 / Fragment: Chorismate mutase / Mutation: E17A, H20L, Q21A, R24I, Q64L, R109N
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579) (bacteria)
Strain: HB8 / ATCC 27634 / DSM 579 / Gene: TTHA0868 / Plasmid: pET29b / Production host: Escherichia coli (E. coli) / Strain (production host): BL21(DE3) / References: UniProt: Q5SJY4
#2: Protein
Divalent-cation tolerance protein CutA


Mass: 12544.401 Da / Num. of mol.: 12 / Fragment: Divalent cation tolerance protein
Mutation: E12A, E13L, R16V, T17K, K20H, W43E, Q44E, E46S, E49S, Q51H, H62D, A73E, T78E
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579) (bacteria)
Strain: HB8 / ATCC 27634 / DSM 579 / Gene: cutA, TTHA1356 / Plasmid: pET29b / Production host: Escherichia coli (E. coli) / Strain (production host): BL21(DE3) / References: UniProt: Q7SIA8

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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: 2.45 Å3/Da / Density % sol: 49.84 %
Crystal growTemperature: 298 K / Method: vapor diffusion, hanging drop / pH: 7.5
Details: 100 mM HEPES pH 7.5, 9% (w/v) PEG 8000, 11.7% (v/v) ethylene glycol

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: APS / Beamline: 24-ID-C / Wavelength: 0.942 Å
DetectorType: DECTRIS PILATUS 6M / Detector: PIXEL / Date: Apr 3, 2014
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.942 Å / Relative weight: 1
ReflectionResolution: 3.25→88.1 Å / Num. obs: 49099 / % possible obs: 96.3 % / Observed criterion σ(I): -3 / Redundancy: 4 % / Biso Wilson estimate: 90.95 Å2 / Rmerge F obs: 0.984 / Rmerge(I) obs: 0.205 / Rrim(I) all: 0.233 / Χ2: 0.917 / Net I/σ(I): 5.54 / Num. measured all: 198097
Reflection shell

Diffraction-ID: 1 / Rejects: 0

Resolution (Å)Rmerge F obsRmerge(I) obsMean I/σ(I) obsNum. measured obsNum. possibleNum. unique obsRrim(I) all% possible all
3.25-3.330.7330.6061.25018372124850.77766.8
3.33-3.430.6870.7481.4310329363033640.89992.7
3.43-3.530.7790.6251.7611685351935080.74299.7
3.53-3.630.8930.5222.6515234342634170.58699.7
3.63-3.750.9130.4423.0814702334233250.49699.5
3.75-3.880.9320.3833.5614011323632180.43199.4
3.88-4.030.9480.3074.3113053312130910.34799
4.03-4.20.9550.2634.9211781299029380.398.3
4.2-4.380.9760.2166.3512801287528650.24199.7
4.38-4.60.9770.2016.9712202276427480.22699.4
4.6-4.840.9790.1777.6711497264426320.19999.5
4.84-5.140.9780.1757.5310656249124790.19799.5
5.14-5.490.9680.1946.459202234623250.22199.1
5.49-5.930.9680.2076.349684219821810.23399.2
5.93-6.50.9720.1926.878958202520110.21599.3
6.5-7.270.9810.1478.57973185918490.16599.5
7.27-8.390.9890.09911.166374164216240.11398.9
8.39-10.280.9930.08114.76062140113840.09198.8
10.28-14.530.9940.06916.134524111710780.07896.5
14.53-88.10.9960.06115.923516575770.06987.8

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Phasing

PhasingMethod: molecular replacement

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Processing

Software
NameVersionClassification
XSCALEdata scaling
PHASER2.5.6phasing
BUSTER2.10.0refinement
PDB_EXTRACT3.15data extraction
XDSdata reduction
RefinementMethod to determine structure: MOLECULAR REPLACEMENT / Resolution: 3.4→88.1 Å / Cor.coef. Fo:Fc: 0.889 / Cor.coef. Fo:Fc free: 0.8487 / Cross valid method: THROUGHOUT / σ(F): 0 / SU Rfree Blow DPI: 0.539
RfactorNum. reflection% reflectionSelection details
Rfree0.2388 2219 5.02 %RANDOM
Rwork0.1895 ---
obs0.192 44218 99.09 %-
Displacement parametersBiso max: 204.2 Å2 / Biso mean: 72.64 Å2 / Biso min: 11.57 Å2
Baniso -1Baniso -2Baniso -3
1--37.3769 Å20 Å20 Å2
2--16.0693 Å20 Å2
3---21.3076 Å2
Refine analyzeLuzzati coordinate error obs: 0.644 Å
Refinement stepCycle: final / Resolution: 3.4→88.1 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms20510 0 0 0 20510
Num. residues----2646
Refine LS restraints
Refine-IDTypeNumberRestraint functionWeightDev ideal
X-RAY DIFFRACTIONt_dihedral_angle_d7454SINUSOIDAL2
X-RAY DIFFRACTIONt_trig_c_planes456HARMONIC2
X-RAY DIFFRACTIONt_gen_planes3022HARMONIC5
X-RAY DIFFRACTIONt_it21042HARMONIC20
X-RAY DIFFRACTIONt_nbd
X-RAY DIFFRACTIONt_improper_torsion
X-RAY DIFFRACTIONt_pseud_angle
X-RAY DIFFRACTIONt_chiral_improper_torsion2850SEMIHARMONIC5
X-RAY DIFFRACTIONt_sum_occupancies
X-RAY DIFFRACTIONt_utility_distance
X-RAY DIFFRACTIONt_utility_angle
X-RAY DIFFRACTIONt_utility_torsion
X-RAY DIFFRACTIONt_ideal_dist_contact25019SEMIHARMONIC4
X-RAY DIFFRACTIONt_bond_d21042HARMONIC20.01
X-RAY DIFFRACTIONt_angle_deg28742HARMONIC21.2
X-RAY DIFFRACTIONt_omega_torsion2.68
X-RAY DIFFRACTIONt_other_torsion18.96
LS refinement shellResolution: 3.4→3.49 Å / Total num. of bins used: 20
RfactorNum. reflection% reflection
Rfree0.3446 158 4.89 %
Rwork0.2416 3076 -
all0.2466 3234 -
obs--99.09 %
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)
10.4210.5109-0.313.00671.59652.39920.03030.1330.0291-0.26390.0203-0.2859-0.19460.1421-0.05060.06-0.00160.0148-0.06520.0004-0.129363.142610.352229.6006
21.9-0.3919-0.83851.8227-0.05920.9564-0.0177-0.0058-0.00820.12150.0834-0.12370.22820.1798-0.06580.1120.0205-0.0422-0.08740.0496-0.121956.7086-2.206645.8041
32.33270.2761-0.74073.60140.29892.1372-0.124-0.0211-0.22590.08710.03670.54390.0705-0.38960.0874-0.1543-0.00230.0342-0.0660.0084-0.0215-1.06394.240642.4488
43.8666-0.52950.44052.05550.03481.49660.0995-0.12630.19750.0584-0.0472-0.0197-0.0264-0.0989-0.05230.07580.0552-0.0096-0.1021-0.0333-0.128353.475819.076547.0088
52.5118-1.36850.23621.8604-0.53040.8175-0.15180.01620.22140.11220.0220.2078-0.1821-0.20.1297-0.0660.00290.0571-0.0579-0.0965-0.01868.078423.456545.9822
60.73050.6978-0.00392.46460.00941.7392-0.1147-0.16710.03620.34770.0410.11970.144-0.12030.07370.00770.06390.0578-0.00730.0165-0.124513.94616.455257.836
72.8160.45940.29370.73360.51860.99940.0793-0.0303-0.2944-0.0792-0.1460.16550.152-0.13330.06670.1683-0.00120.0031-0.09020.0216-0.143423.7443-28.717830.3781
80.7089-0.37980.09385.1902-0.06621.7648-0.05130.2074-0.1483-0.0960.1211-0.02150.2370.2776-0.06980.04840.0049-0.0134-0.0139-0.0385-0.147939.1031-25.430415.4724
91.5801-0.1918-0.54111.20870.26641.98680.04410.0284-0.0465-0.0776-0.02830.2680.2203-0.0303-0.01580.0703-0.0519-0.0828-0.134-0.0359-0.056618.3532-22.051610.5692
100.83910.32160.32931.8086-0.1011.7836-0.00720.17590.056-0.32510.0042-0.0260.07610.27240.0030.19640.00210.0051-0.0567-0.0096-0.175633.48115.1965-3.6266
111.9687-0.57370.14462.6108-0.17252.64920.07120.0740.2028-0.28820.09170.1465-0.11820.2088-0.16290.1097-0.0261-0.0962-0.1218-0.0593-0.133733.150632.49839.3209
122.38090.4469-0.67861.8892-0.70054.2385-0.07220.09210.0993-0.20280.08170.23470.0714-0.2757-0.00940.0160.0399-0.073-0.0822-0.0615-0.114414.651222.66773.9383
132.9743-0.96111.0644.56630.80581.7263-0.01180.3480.0536-0.39530.08110.2258-0.11970.0281-0.06930.0251-0.0598-0.152-0.1145-0.027-0.1254-4.58332.90670.061
143.321-0.8038-0.25570-0.51673.59730.0113-0.3642-0.19240.03690.04910.5376-0.2647-0.3418-0.0604-0.20570.0515-0.0827-0.1258-0.02740.0822-13.4855.14417.485
152.18980.01860.2492.71520.527200.00550.0661-0.2762-0.2040.04540.38820.5032-0.2039-0.0509-0.119-0.051-0.1433-0.1478-0.06220.1129-7.8391-12.698911.5466
160.659-0.31130.25553.10690.34630.8668-0.07470.024-0.34480.20710.07350.0988-0.1121-0.1470.00120.06510.02180.0214-0.04640.0983-0.12236.4085-29.329155.1454
170.87530.17480.3380-0.00190.2676-0.0322-0.3160.07720.3888-0.00830.1982-0.0297-0.27490.04040.20840.00170.0946-0.04430.0722-0.119322.3514-18.784666.1838
182.0730.0833-0.55262.0837-0.24641.47360.0235-0.21840.18160.2692-0.0177-0.4396-0.00440.1643-0.00580.20190.0123-0.0514-0.04510.0072-0.247741.8289-13.647665.7645
191.1933-0.08270.61231.00160.53464.4102-0.00540.44710.0953-0.5394-0.043-0.2089-0.270.08760.04840.1148-0.04730.0871-0.0209-0.0326-0.183359.4945.5725-5.2164
200.0004-0.2345-0.21333.3309-0.30471.4998-0.05540.2834-0.4066-0.54060.1004-0.03680.2847-0.0258-0.0450.2367-0.02070.0188-0.0609-0.101-0.221654.5623-13.359-4.27
212.16220.22271.49152.48110.61561.36190.0553-0.056-0.0783-0.00950.0506-0.54290.14590.4581-0.10590.0520.09110.0706-0.0761-0.0682-0.119368.1333-6.11937.9142
222.45291.8871-0.23260.34030.66682.69130.0363-0.38860.1930.3888-0.03440.3211-0.2256-0.3095-0.00190.0620.01640.014-0.1173-0.0942-0.10727.127443.1849.7406
232.4474-0.4755-0.1612.7371-0.81973.5825-0.09820.36070.2727-0.08770.04540.3666-0.3425-0.20040.05280.09510.0739-0.0849-0.2107-0.0427-0.092627.760948.996631.0341
241.0450.64521.1942.4675-0.43114.0355-0.0067-0.030.0856-0.0139-0.3373-0.3461-0.16020.45920.344-0.0523-0.0326-0.0405-0.07570.0065-0.035344.351144.795740.6323
Refinement TLS group
IDRefine-IDRefine TLS-IDSelection detailsAuth asym-IDAuth seq-ID
1X-RAY DIFFRACTION1{ A|* }A1 - 117
2X-RAY DIFFRACTION2{ B|* }B1 - 117
3X-RAY DIFFRACTION3{ C|* }C1 - 117
4X-RAY DIFFRACTION4{ D|* }D1 - 117
5X-RAY DIFFRACTION5{ E|* }E1 - 117
6X-RAY DIFFRACTION6{ F|* }F1 - 117
7X-RAY DIFFRACTION7{ G|* }G1 - 117
8X-RAY DIFFRACTION8{ H|* }H1 - 117
9X-RAY DIFFRACTION9{ I|* }I1 - 117
10X-RAY DIFFRACTION10{ J|* }J1 - 117
11X-RAY DIFFRACTION11{ K|* }K1 - 117
12X-RAY DIFFRACTION12{ L|* }L1 - 117
13X-RAY DIFFRACTION13{ M|* }M1 - 103
14X-RAY DIFFRACTION14{ N|* }N1 - 103
15X-RAY DIFFRACTION15{ O|* }O1 - 103
16X-RAY DIFFRACTION16{ P|* }P1 - 103
17X-RAY DIFFRACTION17{ Q|* }Q1 - 109
18X-RAY DIFFRACTION18{ R|* }R1 - 103
19X-RAY DIFFRACTION19{ S|* }S1 - 103
20X-RAY DIFFRACTION20{ T|* }T1 - 103
21X-RAY DIFFRACTION21{ U|* }U1 - 103
22X-RAY DIFFRACTION22{ V|* }V1 - 103
23X-RAY DIFFRACTION23{ W|* }W1 - 103
24X-RAY DIFFRACTION24{ X|* }X1 - 103

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