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- PDB-3kqf: 1.8 Angstrom Resolution Crystal Structure of Enoyl-CoA Hydratase ... -

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Basic information

Entry
Database: PDB / ID: 3kqf
Title1.8 Angstrom Resolution Crystal Structure of Enoyl-CoA Hydratase from Bacillus anthracis.
ComponentsEnoyl-CoA hydratase/isomerase family protein
KeywordsISOMERASE / Enoyl-CoA Hydratase / idp02329 / Structural Genomics / Center for Structural Genomics of Infectious Diseases / CSGID
Function / homology
Function and homology information


enoyl-CoA hydratase / isomerase activity
Similarity search - Function
Lyase 2-enoyl-coa Hydratase, Chain A, domain 2 / Lyase 2-enoyl-coa Hydratase; Chain A, domain 2 / Enoyl-CoA hydratase, C-terminal / Enoyl-CoA hydratase/isomerase, conserved site / Enoyl-CoA hydratase/isomerase signature. / Enoyl-CoA hydratase/isomerase / Enoyl-CoA hydratase/isomerase / 2-enoyl-CoA Hydratase; Chain A, domain 1 / 2-enoyl-CoA Hydratase; Chain A, domain 1 / ClpP/crotonase-like domain superfamily ...Lyase 2-enoyl-coa Hydratase, Chain A, domain 2 / Lyase 2-enoyl-coa Hydratase; Chain A, domain 2 / Enoyl-CoA hydratase, C-terminal / Enoyl-CoA hydratase/isomerase, conserved site / Enoyl-CoA hydratase/isomerase signature. / Enoyl-CoA hydratase/isomerase / Enoyl-CoA hydratase/isomerase / 2-enoyl-CoA Hydratase; Chain A, domain 1 / 2-enoyl-CoA Hydratase; Chain A, domain 1 / ClpP/crotonase-like domain superfamily / Alpha-Beta Complex / Orthogonal Bundle / Mainly Alpha / Alpha Beta
Similarity search - Domain/homology
Enoyl-CoA hydratase / Enoyl-CoA hydratase/isomerase family protein
Similarity search - Component
Biological speciesBacillus anthracis (anthrax bacterium)
MethodX-RAY DIFFRACTION / SYNCHROTRON / SAD / Resolution: 1.8 Å
AuthorsMinasov, G. / Halavaty, A. / Wawrzak, Z. / Skarina, T. / Onopriyenko, O. / Papazisi, L. / Savchenko, A. / Anderson, W.F. / Center for Structural Genomics of Infectious Diseases (CSGID)
CitationJournal: TO BE PUBLISHED
Title: 1.8 Angstrom Resolution Crystal Structure of Enoyl-CoA Hydratase from Bacillus anthracis.
Authors: Minasov, G. / Halavaty, A. / Wawrzak, Z. / Skarina, T. / Onopriyenko, O. / Papazisi, L. / Savchenko, A. / Anderson, W.F.
History
DepositionNov 17, 2009Deposition site: RCSB / Processing site: RCSB
Revision 1.0Nov 24, 2009Provider: repository / Type: Initial release
Revision 1.1Jul 13, 2011Group: Advisory / Version format compliance
Revision 1.2Nov 1, 2017Group: Refinement description / Category: software / Item: _software.name

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

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Assembly

Deposited unit
A: Enoyl-CoA hydratase/isomerase family protein
B: Enoyl-CoA hydratase/isomerase family protein
C: Enoyl-CoA hydratase/isomerase family protein
D: Enoyl-CoA hydratase/isomerase family protein
E: Enoyl-CoA hydratase/isomerase family protein
F: Enoyl-CoA hydratase/isomerase family protein
hetero molecules


Theoretical massNumber of molelcules
Total (without water)174,84913
Polymers174,5786
Non-polymers2717
Water15,187843
1
A: Enoyl-CoA hydratase/isomerase family protein
B: Enoyl-CoA hydratase/isomerase family protein
C: Enoyl-CoA hydratase/isomerase family protein
hetero molecules


Theoretical massNumber of molelcules
Total (without water)87,3294
Polymers87,2893
Non-polymers401
Water543
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area11250 Å2
ΔGint-73 kcal/mol
Surface area29710 Å2
MethodPISA
2
D: Enoyl-CoA hydratase/isomerase family protein
E: Enoyl-CoA hydratase/isomerase family protein
F: Enoyl-CoA hydratase/isomerase family protein
hetero molecules


Theoretical massNumber of molelcules
Total (without water)87,5209
Polymers87,2893
Non-polymers2316
Water543
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area11600 Å2
ΔGint-114 kcal/mol
Surface area30000 Å2
MethodPISA
Unit cell
Length a, b, c (Å)73.747, 130.721, 73.802
Angle α, β, γ (deg.)90.00, 114.47, 90.00
Int Tables number4
Space group name H-MP1211

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Components

#1: Protein
Enoyl-CoA hydratase/isomerase family protein


Mass: 29096.311 Da / Num. of mol.: 6
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Bacillus anthracis (anthrax bacterium) / Gene: BAS2374, BA_2551, GBAA2551, GBAA_2551 / Plasmid: pMCSG7 / Production host: Escherichia coli (E. coli) / Strain (production host): BL21-DE3
References: UniProt: Q81Q82, UniProt: A0A6L8PDF4*PLUS, enoyl-CoA hydratase
#2: Chemical
ChemComp-CA / CALCIUM ION


Mass: 40.078 Da / Num. of mol.: 5 / Source method: obtained synthetically / Formula: Ca
#3: Chemical ChemComp-CL / CHLORIDE ION


Mass: 35.453 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: Cl
#4: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 843 / 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: 1.85 Å3/Da / Density % sol: 33.68 %
Crystal growTemperature: 295 K / Method: vapor diffusion, hanging drop
Details: Protein solution: 0.3M NaCl, 10mM HEPES, pH 7.5. Screen solution: PEG400 28%, CaCl2 0.2M, Hepes 0.1M, pH 7.5, VAPOR DIFFUSION, HANGING DROP, temperature 295K

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: APS / Beamline: 21-ID-G / Wavelength: 0.97856 Å
DetectorType: MARMOSAIC 300 mm CCD / Detector: CCD / Date: Oct 29, 2009 / Details: Beryllium lenses
RadiationMonochromator: Diamond / Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.97856 Å / Relative weight: 1
ReflectionResolution: 1.8→30 Å / Num. all: 116373 / Num. obs: 116373 / % possible obs: 100 % / Observed criterion σ(I): -3 / Redundancy: 3.8 % / Biso Wilson estimate: 22.1 Å2 / Rmerge(I) obs: 0.091 / Net I/σ(I): 14.3
Reflection shellResolution: 1.8→1.83 Å / Redundancy: 3.7 % / Rmerge(I) obs: 0.556 / Mean I/σ(I) obs: 2.3 / Num. unique all: 5755 / % possible all: 100

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Processing

Software
NameVersionClassification
Blu-IceMaxdata collection
CRANKphasing
REFMAC5.5.0102refinement
HKL-2000data reduction
HKL-2000data scaling
RefinementMethod to determine structure: SAD / Resolution: 1.8→29.87 Å / Cor.coef. Fo:Fc: 0.969 / Cor.coef. Fo:Fc free: 0.957 / SU B: 4.048 / SU ML: 0.058 / TLS residual ADP flag: LIKELY RESIDUAL
Isotropic thermal model: Individual Temperature Factors Refined
Cross valid method: THROUGHOUT / ESU R: 0.139 / ESU R Free: 0.124 / Stereochemistry target values: MAXIMUM LIKELIHOOD / Details: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS
RfactorNum. reflection% reflectionSelection details
Rfree0.19325 5782 5 %RANDOM
Rwork0.1555 ---
all0.15737 109827 --
obs0.15737 109827 98.82 %-
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: 15.177 Å2
Baniso -1Baniso -2Baniso -3
1--1.23 Å20 Å20.98 Å2
2--0.66 Å2-0 Å2
3---1.38 Å2
Refinement stepCycle: LAST / Resolution: 1.8→29.87 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms11812 0 7 843 12662
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.0110.02212628
X-RAY DIFFRACTIONr_bond_other_d0.0010.028550
X-RAY DIFFRACTIONr_angle_refined_deg1.3241.98217145
X-RAY DIFFRACTIONr_angle_other_deg0.864321090
X-RAY DIFFRACTIONr_dihedral_angle_1_deg2.70451689
X-RAY DIFFRACTIONr_dihedral_angle_2_deg33.08224.991541
X-RAY DIFFRACTIONr_dihedral_angle_3_deg9.265152385
X-RAY DIFFRACTIONr_dihedral_angle_4_deg12.1981592
X-RAY DIFFRACTIONr_chiral_restr0.080.22002
X-RAY DIFFRACTIONr_gen_planes_refined0.0050.0214417
X-RAY DIFFRACTIONr_gen_planes_other0.0010.022319
X-RAY DIFFRACTIONr_mcbond_it0.9251.58116
X-RAY DIFFRACTIONr_mcbond_other0.3091.53330
X-RAY DIFFRACTIONr_mcangle_it1.56213117
X-RAY DIFFRACTIONr_scbond_it2.78534512
X-RAY DIFFRACTIONr_scangle_it4.5714.54028
LS refinement shellResolution: 1.8→1.849 Å / Total num. of bins used: 20
RfactorNum. reflection% reflection
Rfree0.257 399 -
Rwork0.204 7450 -
obs-7450 91.15 %
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)
12.2962-0.53990.74311.0691-0.23592.17170.03870.30220.009-0.1407-0.02-0.17630.00020.3202-0.01870.078-0.01570.0510.1307-0.03410.07749.331231.3897-33.2673
24.1207-0.88832.38920.2394-0.43011.9964-0.098-0.06660.115-0.03050.0996-0.0731-0.12880.1819-0.00150.1404-0.05010.0110.1826-0.06010.16410.961339.4144-19.4477
30.61120.09010.53051.22460.67831.896-0.02430.0605-0.17040.00240.0503-0.14920.11310.0614-0.0260.0579-0.01020.0250.0386-0.02390.08990.492526.0505-24.2421
40.5205-0.12670.12150.47510.09361.3801-0.01830.0645-0.1497-0.03920.0132-0.04230.1273-0.02580.00510.0756-0.00170.00890.02-0.02520.063-3.367225.5386-20.7003
51.0002-0.3033-0.30871.2930.65783.42010.06990.2166-0.0023-0.2218-0.10960.0793-0.0833-0.11510.03970.1101-0.0007-0.01780.0626-0.01590.0378-16.72941.2435-39.6509
62.39210.2372-0.77831.36820.28792.351-0.02170.30320.1424-0.280.02060.2642-0.0633-0.29850.00110.0941-0.0157-0.08090.1585-0.03050.1186-42.138133.73-28.3246
73.15460.4015-1.45283.2999-0.68248.6025-0.16040.79810.3028-0.9756-0.09710.042-0.4138-0.07290.25750.38880.0042-0.06660.27060.0510.193-31.845143.4637-35.3011
81.33940.303-0.19051.31460.0241.7584-0.05230.1416-0.1041-0.2201-0.02510.00320.1031-0.11180.07740.0809-0.0214-0.00090.0628-0.04740.0506-29.848628.2611-27.2864
91.20940.3871-0.65260.38420.1481.0926-0.04490.0562-0.1163-0.0383-0.03920.03350.0973-0.11390.08420.0834-0.0104-0.01460.0487-0.03630.0676-24.775227.2991-25.7491
102.96211.1395-0.44811.68320.13521.96830.1103-0.2070.08360.2133-0.21090.22090.0696-0.33780.10060.1058-0.02380.02110.0923-0.04290.0569-34.478638.1847-1.7692
113.65040.3816-0.78641.13870.19611.87930.015-0.4831-0.17570.2413-0.0321-0.03660.07430.07320.01710.19560.0197-0.00150.11420.06030.0375-12.810427.427612.6384
129.99470.0608-4.10970.3190.28492.7450.2872-0.0720.27610.104-0.12310.1027-0.2423-0.3247-0.16410.21450.04320.03430.1508-0.00630.0934-23.499138.39098.8234
131.9629-0.5902-0.10271.00170.26641.6355-0.0053-0.1816-0.21970.135-0.01610.15150.1955-0.09060.02150.1252-0.01580.01780.02720.01520.0412-17.442624.45240.1127
142.328-0.13140.26720.6615-0.10070.5824-0.0256-0.0719-0.25830.0602-0.0280.01230.1769-0.04220.05370.1111-0.0160.0210.00650.00240.044-18.57424.6742-4.8724
152.0081-0.5680.38791.9005-0.16551.2268-0.0432-0.0624-0.09210.13580.0133-0.2321-0.080.17310.02990.0734-0.0062-0.02630.0457-0.00860.06837.474635.6301-5.5874
161.55630.04680.67431.0907-0.47733.5296-0.02390.2357-0.0226-0.1214-0.0256-0.19920.04620.38650.04960.0709-0.02040.05090.1231-0.03130.099512.527365.7217-28.2123
170.5364-0.13940.80410.2737-1.2636.77270.00740.2054-0.0747-0.05430.0667-0.050.2019-0.0536-0.07410.1976-0.03380.02860.1971-0.07870.1440.777157.6808-35.4355
181.67320.56470.97321.32910.60941.3473-0.03460.05660.1357-0.17440.0501-0.0527-0.15710.0607-0.01540.058-0.01540.02630.0121-0.00390.02840.684571.0263-23.851
191.30050.09960.47060.6027-0.11240.61280.0119-0.04050.1512-0.02230.013-0.0501-0.09950.0426-0.02490.087-0.01270.01820.0258-0.01480.0517-4.252871.5298-21.9223
203.79970.96730.81311.3380.17811.44660.052-0.2338-0.14230.1454-0.0972-0.20960.02060.15730.04520.0957-0.006-0.03410.06490.00390.07077.788456.0316-1.7665
212.58530.3559-0.32731.59850.52981.7839-0.0048-0.4276-0.06930.36450.0278-0.16330.16390.1157-0.0230.2236-0.0203-0.02740.187-0.03440.0307-13.254363.279816.7831
227.6884-0.59740.63794.00380.98212.66380.1453-0.4002-0.46580.54360.1245-1.04290.48330.6277-0.26980.31420.0601-0.11270.26820.00670.3434-2.992153.556810.0972
232.02810.35970.18441.34910.2061.3530.0829-0.20.16660.0554-0.0524-0.1862-0.04770.0724-0.03050.1026-0.0254-0.00380.0646-0.03880.0669-9.149568.79054.973
241.7329-0.1866-0.23940.52030.44150.43970.0508-0.17080.16990.0458-0.0491-0.0081-0.0399-0.019-0.00180.1003-0.0296-0.00860.0406-0.03050.0401-8.484569.7621-0.2902
253.1111-0.4937-0.68021.58861.2712.64860.0519-0.33030.09420.0629-0.2240.32650.0306-0.33580.17210.097-0.03120.01330.1141-0.05540.106-34.478159.0903-1.169
262.74070.051-1.13341.06380.07782.21410.00880.28360.0867-0.1008-0.05340.2457-0.1192-0.39940.04460.08660.0385-0.05430.11680.03380.1491-38.506569.6507-27.0343
276.63240.7082-5.20720.1952-0.23385.9542-0.1494-0.2397-0.4189-0.0149-0.13560.0510.0997-0.17460.2850.1271-0.0135-0.00310.170.01230.2428-39.404758.6954-15.717
281.68160.07860.1070.719-0.45051.52370.0134-0.01730.28780.0695-0.03630.1167-0.2002-0.13280.02290.09510.0138-0.01620.0218-0.01260.1032-28.911372.5654-17.7144
290.7068-0.0033-0.40770.42370.03851.36170.0429-0.00910.2415-0.0457-0.01390.0558-0.1477-0.0956-0.0290.08230.0046-0.00530.0127-0.01510.1006-24.901972.4528-14.5624
301.1891-0.11260.32931.8602-0.62132.21660.05980.2222-0.031-0.29650.00220.0303-0.00950.0211-0.0620.1201-0.0136-0.00420.0552-0.01260.0176-13.401161.5996-38.1553
Refinement TLS group
IDRefine-IDRefine TLS-IDAuth asym-IDAuth seq-ID
1X-RAY DIFFRACTION1A1 - 67
2X-RAY DIFFRACTION2A68 - 100
3X-RAY DIFFRACTION3A101 - 134
4X-RAY DIFFRACTION4A135 - 204
5X-RAY DIFFRACTION5A205 - 262
6X-RAY DIFFRACTION6B1 - 67
7X-RAY DIFFRACTION7B68 - 100
8X-RAY DIFFRACTION8B101 - 134
9X-RAY DIFFRACTION9B135 - 204
10X-RAY DIFFRACTION10B205 - 262
11X-RAY DIFFRACTION11C1 - 67
12X-RAY DIFFRACTION12C68 - 100
13X-RAY DIFFRACTION13C101 - 134
14X-RAY DIFFRACTION14C135 - 204
15X-RAY DIFFRACTION15C205 - 262
16X-RAY DIFFRACTION16D1 - 67
17X-RAY DIFFRACTION17D68 - 100
18X-RAY DIFFRACTION18D101 - 134
19X-RAY DIFFRACTION19D135 - 204
20X-RAY DIFFRACTION20D205 - 262
21X-RAY DIFFRACTION21E1 - 67
22X-RAY DIFFRACTION22E68 - 100
23X-RAY DIFFRACTION23E101 - 134
24X-RAY DIFFRACTION24E135 - 204
25X-RAY DIFFRACTION25E205 - 262
26X-RAY DIFFRACTION26F1 - 67
27X-RAY DIFFRACTION27F68 - 100
28X-RAY DIFFRACTION28F101 - 134
29X-RAY DIFFRACTION29F135 - 204
30X-RAY DIFFRACTION30F205 - 262

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