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- PDB-3n8k: Type II dehydroquinase from Mycobacterium tuberculosis complexed ... -

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Basic information

Entry
Database: PDB / ID: 3n8k
TitleType II dehydroquinase from Mycobacterium tuberculosis complexed with citrazinic acid
Components3-dehydroquinate dehydratase
KeywordsLYASE / DEHYDRATASE / SHIKIMATE PATHWAY / AROMATIC AMINO ACID BIOSYNTHESIS / tuberculosis / drug target / citrazinic acid / Structural Genomics / TB Structural Genomics Consortium / TBSGC
Function / homology
Function and homology information


quinate catabolic process / Chorismate via Shikimate Pathway / 3-dehydroquinate dehydratase / 3-dehydroquinate dehydratase activity / chorismate biosynthetic process / aromatic amino acid family biosynthetic process / amino acid biosynthetic process / cytosol
Similarity search - Function
Dehydroquinase, class II / Dehydroquinase, class II, conserved site / Dehydroquinase class II signature. / Dehydroquinase, class II / Dehydroquinase, class II superfamily / Dehydroquinase class II / Rossmann fold / 3-Layer(aba) Sandwich / Alpha Beta
Similarity search - Domain/homology
Chem-D1X / 3-dehydroquinate dehydratase / 3-dehydroquinate dehydratase
Similarity search - Component
Biological speciesMycobacterium tuberculosis (bacteria)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 2.25 Å
AuthorsSnee, W.C. / Palaninathan, S.K. / Sacchettini, J.C. / Dias, M.V.B. / Bromfield, K.M. / Payne, R. / Ciulli, A. / Howard, N.I. / Abell, C. / Blundell, T.L. / TB Structural Genomics Consortium (TBSGC)
CitationJournal: Biochem.J. / Year: 2011
Title: Structural investigation of inhibitor designs targeting 3-dehydroquinate dehydratase from the shikimate pathway of Mycobacterium tuberculosis.
Authors: Dias, M.V. / Snee, W.C. / Bromfield, K.M. / Payne, R.J. / Palaninathan, S.K. / Ciulli, A. / Howard, N.I. / Abell, C. / Sacchettini, J.C. / Blundell, T.L.
History
DepositionMay 28, 2010Deposition site: RCSB / Processing site: RCSB
Revision 1.0Jul 21, 2010Provider: repository / Type: Initial release
Revision 1.1Jul 13, 2011Group: Version format compliance
Revision 1.2Sep 6, 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 / struct_ref_seq_dif / struct_site
Item: _database_2.pdbx_DOI / _database_2.pdbx_database_accession ..._database_2.pdbx_DOI / _database_2.pdbx_database_accession / _struct_ref_seq_dif.details / _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: 3-dehydroquinate dehydratase
B: 3-dehydroquinate dehydratase
C: 3-dehydroquinate dehydratase
D: 3-dehydroquinate dehydratase
E: 3-dehydroquinate dehydratase
F: 3-dehydroquinate dehydratase
G: 3-dehydroquinate dehydratase
H: 3-dehydroquinate dehydratase
I: 3-dehydroquinate dehydratase
J: 3-dehydroquinate dehydratase
K: 3-dehydroquinate dehydratase
L: 3-dehydroquinate dehydratase
M: 3-dehydroquinate dehydratase
N: 3-dehydroquinate dehydratase
O: 3-dehydroquinate dehydratase
P: 3-dehydroquinate dehydratase
Q: 3-dehydroquinate dehydratase
R: 3-dehydroquinate dehydratase
S: 3-dehydroquinate dehydratase
T: 3-dehydroquinate dehydratase
U: 3-dehydroquinate dehydratase
V: 3-dehydroquinate dehydratase
W: 3-dehydroquinate dehydratase
X: 3-dehydroquinate dehydratase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)451,93372
Polymers447,59924
Non-polymers4,33448
Water23,5461307
1
A: 3-dehydroquinate dehydratase
B: 3-dehydroquinate dehydratase
C: 3-dehydroquinate dehydratase
D: 3-dehydroquinate dehydratase
E: 3-dehydroquinate dehydratase
F: 3-dehydroquinate dehydratase
G: 3-dehydroquinate dehydratase
H: 3-dehydroquinate dehydratase
I: 3-dehydroquinate dehydratase
J: 3-dehydroquinate dehydratase
K: 3-dehydroquinate dehydratase
L: 3-dehydroquinate dehydratase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)226,33443
Polymers223,79912
Non-polymers2,53531
Water21612
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area30430 Å2
ΔGint-233 kcal/mol
Surface area52370 Å2
MethodPISA
2
M: 3-dehydroquinate dehydratase
N: 3-dehydroquinate dehydratase
O: 3-dehydroquinate dehydratase
P: 3-dehydroquinate dehydratase
Q: 3-dehydroquinate dehydratase
R: 3-dehydroquinate dehydratase
S: 3-dehydroquinate dehydratase
T: 3-dehydroquinate dehydratase
U: 3-dehydroquinate dehydratase
V: 3-dehydroquinate dehydratase
W: 3-dehydroquinate dehydratase
X: 3-dehydroquinate dehydratase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)225,59929
Polymers223,79912
Non-polymers1,79917
Water21612
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area28870 Å2
ΔGint-125 kcal/mol
Surface area54370 Å2
MethodPISA
Unit cell
Length a, b, c (Å)96.316, 137.208, 146.713
Angle α, β, γ (deg.)90.00, 96.59, 90.00
Int Tables number4
Space group name H-MP1211

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Components

#1: Protein ...
3-dehydroquinate dehydratase / 3-dehydroquinase / Type II DHQase


Mass: 18649.947 Da / Num. of mol.: 24
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Mycobacterium tuberculosis (bacteria) / Gene: aroD, aroQ, MT2612, MTCY159.19, Rv2537c / Production host: Escherichia coli (E. coli) / Strain (production host): BL21(DE3)
References: UniProt: P0A4Z6, UniProt: P9WPX7*PLUS, 3-dehydroquinate dehydratase
#2: Chemical...
ChemComp-D1X / 2,6-dioxo-1,2,3,6-tetrahydropyridine-4-carboxylic acid


Mass: 155.108 Da / Num. of mol.: 22 / Source method: obtained synthetically / Formula: C6H5NO4
#3: Chemical...
ChemComp-CL / CHLORIDE ION


Mass: 35.453 Da / Num. of mol.: 26 / Source method: obtained synthetically / Formula: Cl
#4: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 1307 / 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: 2.15 Å3/Da / Density % sol: 42.83 %
Crystal growTemperature: 290 K / Method: microbatch
Details: MtDHQase protein was concentrated to 10-15 mg/mL prior to crystallization, using Amicon Ultra MWCO 5 KDa protein concentrators. MtDHQase crystals were grown in 4 uL drops composed of 5-7 ...Details: MtDHQase protein was concentrated to 10-15 mg/mL prior to crystallization, using Amicon Ultra MWCO 5 KDa protein concentrators. MtDHQase crystals were grown in 4 uL drops composed of 5-7 mg/mL protein, 15% PEG monomethyl ether 2,000, 0.075 M KBr, 25 mM Tris, 50 mM NaCl, 0.5 mM DTT, 0.5 mM EDTA, pH 7.5 in microbatch plates covered with 5 mL Al s oil, Microbatch, temperature 290K

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: APS / Beamline: 23-ID-B / Wavelength: 0.97934 Å
DetectorType: MARMOSAIC 300 mm CCD / Detector: CCD / Date: Mar 14, 2008
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.97934 Å / Relative weight: 1
ReflectionResolution: 1.77→50 Å / Num. all: 176635 / Num. obs: 176635 / % possible obs: 98.44 % / Observed criterion σ(F): -3 / Observed criterion σ(I): -3

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Processing

Software
NameVersionClassification
HKL-3000data collection
MOLREPphasing
REFMAC5.2.0019refinement
HKL-2000data reduction
HKL-2000data scaling
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: PDB entry 3N59
Resolution: 2.25→50 Å / Cor.coef. Fo:Fc: 0.937 / Cor.coef. Fo:Fc free: 0.916 / SU B: 14.791 / SU ML: 0.17 / Cross valid method: THROUGHOUT / ESU R: 0.32 / ESU R Free: 0.219 / Stereochemistry target values: MAXIMUM LIKELIHOOD / Details: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS
RfactorNum. reflection% reflectionSelection details
Rfree0.23612 8851 5 %RANDOM
Rwork0.20479 ---
obs0.20636 167784 98.43 %-
Solvent computationIon probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.4 Å / Solvent model: MASK
Displacement parametersBiso mean: 43.441 Å2
Baniso -1Baniso -2Baniso -3
1--0.25 Å20 Å20.21 Å2
2--0.33 Å20 Å2
3----0.03 Å2
Refinement stepCycle: LAST / Resolution: 2.25→50 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms25582 0 268 1307 27157
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.0050.02126285
X-RAY DIFFRACTIONr_bond_other_d
X-RAY DIFFRACTIONr_angle_refined_deg0.8811.9735816
X-RAY DIFFRACTIONr_angle_other_deg
X-RAY DIFFRACTIONr_dihedral_angle_1_deg4.43553375
X-RAY DIFFRACTIONr_dihedral_angle_2_deg33.25223.3711160
X-RAY DIFFRACTIONr_dihedral_angle_3_deg14.404154103
X-RAY DIFFRACTIONr_dihedral_angle_4_deg13.44215228
X-RAY DIFFRACTIONr_chiral_restr0.0540.24218
X-RAY DIFFRACTIONr_gen_planes_refined0.0030.02120043
X-RAY DIFFRACTIONr_gen_planes_other
X-RAY DIFFRACTIONr_nbd_refined0.1590.212675
X-RAY DIFFRACTIONr_nbd_other
X-RAY DIFFRACTIONr_nbtor_refined0.290.217350
X-RAY DIFFRACTIONr_nbtor_other
X-RAY DIFFRACTIONr_xyhbond_nbd_refined0.0880.21820
X-RAY DIFFRACTIONr_xyhbond_nbd_other
X-RAY DIFFRACTIONr_metal_ion_refined
X-RAY DIFFRACTIONr_metal_ion_other
X-RAY DIFFRACTIONr_symmetry_vdw_refined0.150.265
X-RAY DIFFRACTIONr_symmetry_vdw_other
X-RAY DIFFRACTIONr_symmetry_hbond_refined0.0770.227
X-RAY DIFFRACTIONr_symmetry_hbond_other
X-RAY DIFFRACTIONr_symmetry_metal_ion_refined
X-RAY DIFFRACTIONr_symmetry_metal_ion_other
X-RAY DIFFRACTIONr_mcbond_it0.1171.516767
X-RAY DIFFRACTIONr_mcbond_other
X-RAY DIFFRACTIONr_mcangle_it0.229226718
X-RAY DIFFRACTIONr_scbond_it0.36239518
X-RAY DIFFRACTIONr_scangle_it0.6434.59093
X-RAY DIFFRACTIONr_rigid_bond_restr
X-RAY DIFFRACTIONr_sphericity_free
X-RAY DIFFRACTIONr_sphericity_bonded
LS refinement shellResolution: 2.25→2.307 Å / Total num. of bins used: 20
RfactorNum. reflection% reflection
Rfree0.291 616 -
Rwork0.257 11671 -
obs--93.16 %
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)
12.5836-0.05310.92932.1552-0.05392.0253-0.0463-0.1687-0.17010.02640.02060.00530.1547-0.05050.02570.1033-0.0180.02660.03860.04080.059-35.5423.0675-15.4879
22.41170.46110.09982.33980.29181.8645-0.03470.22550.0476-0.24430.0878-0.0822-0.10230.0201-0.05310.0623-0.05640.02820.13550.03750.0807-20.935844.5889-46.3801
32.1737-0.597-0.52731.90960.4792.19330.0401-0.15240.3247-0.0025-0.0214-0.1947-0.23740.1479-0.01870.1069-0.06170.01450.0808-0.02780.2204-12.430560.2113-21.4916
41.73370.058-0.01871.90030.30192.7835-0.0072-0.01370.13680.04770.00650.1841-0.0244-0.14970.00070.0005-0.00020.00910.0658-0.03870.0981-62.374640.1054-25.5181
51.7729-0.0437-0.04372.2333-0.08042.4185-0.0358-0.0357-0.0434-0.00560.0167-0.24630.04690.20610.01910.0065-0.01970.00760.11660.02150.1752-1.190132.447-26.6325
61.8630.49650.20662.2622-0.30731.8018-0.03820.13060.0956-0.130.0275-0.01220.02080.05540.01070.0158-0.0152-0.0070.0920.02260.0351-42.437935.7171-47.9618
72.6331-0.81570.36782.0216-0.26461.89020.03890.0405-0.2031-0.0238-0.09950.11290.1864-0.12940.06050.0575-0.02710.00260.0463-0.02960.0474-50.989112.4051-30.2445
82.4859-0.2012-0.26511.6677-0.02271.84350.0318-0.2160.33990.0958-0.03-0.114-0.2520.112-0.00180.1592-0.0730.01730.1171-0.12420.1983-28.23463.6015-4.5152
91.95720.26460.18891.95910.57962.7996-0.0053-0.08770.19420.1218-0.02780.2365-0.1046-0.12580.03310.0740.00680.03550.0775-0.09490.1531-56.005951.6744-6.2155
101.5879-0.21470.28142.91990.03271.27970.0456-0.35970.05730.3396-0.01480.07130.02470.0458-0.03080.242-0.02630.0180.2273-0.07390.0571-36.186740.235113.4134
112.31880.7466-0.35592.1732-0.78842.7283-0.0844-0.1166-0.27030.0619-0.0311-0.44040.18780.2620.11550.06350.023-0.00350.12590.04510.2264-7.688614.6995-12.6804
122.22180.1238-0.25983.11730.3841.55550.0799-0.3965-0.02290.39540.0105-0.11710.08620.1157-0.09030.2394-0.0001-0.06320.19270.01910.0216-27.807818.78399.5193
131.9648-0.057-1.19441.30861.26884.24670.37060.01280.363-0.00110.2955-0.2067-0.54830.2769-0.66610.28210.00410.18830.1211-0.12510.3526-18.2621136.2236-47.231
142.1687-1.011-0.39492.38060.67493.4056-0.3833-0.2305-0.21520.71370.34760.14951.36860.03940.03571.25590.2154-0.0630.20050.08490.1576-20.35382.6328-47.2927
152.7492-0.4092-0.08391.72990.03713.6307-0.4355-0.5381-0.18920.54270.4432-0.27231.03160.9612-0.00771.20210.8509-0.30220.8815-0.1820.33119.69180.6227-45.0813
162.23130.0556-0.52331.73540.04843.44140.0768-0.04380.1470.02270.2222-0.09720.18730.0925-0.2990.03440.0258-0.0440.1139-0.0370.1016-12.4996108.7779-85.3723
173.26750.5237-0.70192.21760.43843.6222-0.0201-0.71930.0050.7820.4708-0.2171.17390.9623-0.45071.12640.6391-0.39210.7223-0.1930.2464-6.18697.3507-24.966
182.2426-0.7514-0.40392.20110.29513.6171-0.08960.0665-0.17950.30990.07740.13320.953-0.38930.01220.452-0.0994-0.05060.16680.06740.1123-29.995194.8652-64.7553
191.7274-0.1631-0.41342.27550.5164.1520.1990.14510.30940.07270.1310.0458-0.302-0.5155-0.330.12770.12640.09110.17870.06680.188-28.3161125.1016-65.3837
201.5744-0.689-0.88131.8037-0.81763.4509-0.2341-0.66160.01120.4970.463-0.56570.78731.429-0.22890.68710.7741-0.41151.3075-0.46810.514623.35191.4536-61.1712
213.2531-0.4059-1.25242.12011.0673.59370.0311-0.52810.40180.02540.5533-0.45950.13050.8495-0.58440.03440.1073-0.05240.4746-0.33730.319910.9094107.8811-83.7023
220.19410.44030.63815.01560.35663.48960.2231-0.38950.3986-0.1507-0.0437-1.5659-0.13352.157-0.17930.122-0.07260.10472.2645-0.96871.385225.524121.8579-61.1648
231.46031.1412-0.09542.5818-0.34383.33040.2158-0.49360.22330.52270.4477-0.0990.41560.8888-0.66350.44350.3138-0.17690.861-0.48860.4488-3.7829120.8592-25.4891
242.0279-1.0329-1.26181.21130.60543.59480.3058-0.64950.64350.07210.5592-0.6784-0.59531.5756-0.86490.3083-0.26290.13551.1934-0.86250.880312.8495135.907-46.8585
Refinement TLS group
IDRefine-IDRefine TLS-IDAuth asym-IDAuth seq-ID
1X-RAY DIFFRACTION1A3 - 144
2X-RAY DIFFRACTION2B3 - 144
3X-RAY DIFFRACTION3C2 - 143
4X-RAY DIFFRACTION4D2 - 144
5X-RAY DIFFRACTION5E2 - 144
6X-RAY DIFFRACTION6F3 - 143
7X-RAY DIFFRACTION7G2 - 144
8X-RAY DIFFRACTION8H3 - 143
9X-RAY DIFFRACTION9I3 - 143
10X-RAY DIFFRACTION10J3 - 143
11X-RAY DIFFRACTION11K3 - 143
12X-RAY DIFFRACTION12L3 - 143
13X-RAY DIFFRACTION13M-6 - 144
14X-RAY DIFFRACTION14N3 - 143
15X-RAY DIFFRACTION15O3 - 143
16X-RAY DIFFRACTION16P3 - 143
17X-RAY DIFFRACTION17Q3 - 143
18X-RAY DIFFRACTION18R3 - 143
19X-RAY DIFFRACTION19S3 - 143
20X-RAY DIFFRACTION20T3 - 143
21X-RAY DIFFRACTION21U3 - 143
22X-RAY DIFFRACTION22V3 - 143
23X-RAY DIFFRACTION23W3 - 143
24X-RAY DIFFRACTION24X3 - 143

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