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- PDB-9f8o: Crystal Structure of PhzA/B from Burkholderia cepacia R18194 in c... -

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Basic information

Entry
Database: PDB / ID: 9f8o
TitleCrystal Structure of PhzA/B from Burkholderia cepacia R18194 in complex with [6-Hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophen-3-yl]{2-[(2-morpholinoethyl)amino]phenyl}methanone
ComponentsPhenazine biosynthesis protein A/B
KeywordsBIOSYNTHETIC PROTEIN / pyocyanin / phenazine biosynthesis / virulence / inhibitor / ketosteroid-isomerase / cocrystal / PhzA/B / Burkholderia cepacia
Function / homologyPhenazine biosynthesis protein A/B / Phenazine biosynthesis protein A/B / antibiotic biosynthetic process / NTF2-like domain superfamily / : / ACETATE ION / Phenazine biosynthesis protein A/B
Function and homology information
Biological speciesBurkholderia lata (bacteria)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 1.57 Å
AuthorsThiemann, M. / Zimmermann, M. / Kunick, C. / Blankenfeldt, W.
Funding support1items
OrganizationGrant numberCountry
Not funded
CitationJournal: J.Med.Chem. / Year: 2025
Title: From Bones to Bugs: Structure-Based Development of Raloxifene-Derived Pathoblockers That Inhibit Pyocyanin Production in Pseudomonas aeruginosa.
Authors: Thiemann, M. / Zimmermann, M. / Diederich, C. / Zhan, H. / Lebedev, M. / Pletz, J. / Baumgarten, J. / Handke, M. / Musken, M. / Breinbauer, R. / Krasteva-Christ, G. / Zanin, E. / Empting, M. ...Authors: Thiemann, M. / Zimmermann, M. / Diederich, C. / Zhan, H. / Lebedev, M. / Pletz, J. / Baumgarten, J. / Handke, M. / Musken, M. / Breinbauer, R. / Krasteva-Christ, G. / Zanin, E. / Empting, M. / Schiedel, M. / Kunick, C. / Blankenfeldt, W.
History
DepositionMay 6, 2024Deposition site: PDBE / Processing site: PDBE
Revision 1.0Apr 16, 2025Provider: repository / Type: Initial release

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: Phenazine biosynthesis protein A/B
B: Phenazine biosynthesis protein A/B
C: Phenazine biosynthesis protein A/B
D: Phenazine biosynthesis protein A/B
hetero molecules


Theoretical massNumber of molelcules
Total (without water)88,81118
Polymers86,1244
Non-polymers2,68714
Water9,476526
1
A: Phenazine biosynthesis protein A/B
B: Phenazine biosynthesis protein A/B
hetero molecules


Theoretical massNumber of molelcules
Total (without water)44,4069
Polymers43,0622
Non-polymers1,3447
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area5450 Å2
ΔGint-23 kcal/mol
Surface area13960 Å2
MethodPISA
2
C: Phenazine biosynthesis protein A/B
D: Phenazine biosynthesis protein A/B
hetero molecules


Theoretical massNumber of molelcules
Total (without water)44,4069
Polymers43,0622
Non-polymers1,3447
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area5460 Å2
ΔGint-22 kcal/mol
Surface area13850 Å2
MethodPISA
Unit cell
Length a, b, c (Å)68.830, 69.629, 83.141
Angle α, β, γ (deg.)90.00, 90.12, 90.00
Int Tables number4
Space group name H-MP1211

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Components

#1: Protein
Phenazine biosynthesis protein A/B


Mass: 21531.025 Da / Num. of mol.: 4
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Burkholderia lata (bacteria) / Gene: Bcep18194_B1568 / Plasmid: pET15b / Production host: Escherichia coli BL21(DE3) (bacteria) / References: UniProt: Q396C9
#2: Chemical
ChemComp-ACT / ACETATE ION


Mass: 59.044 Da / Num. of mol.: 4 / Source method: obtained synthetically / Formula: C2H3O2
#3: Chemical
ChemComp-GOL / GLYCEROL / GLYCERIN / PROPANE-1,2,3-TRIOL


Mass: 92.094 Da / Num. of mol.: 6 / Source method: obtained synthetically / Formula: C3H8O3
#4: Chemical
ChemComp-A1IAF / [2-(4-hydroxyphenyl)-6-oxidanyl-1-benzothiophen-3-yl]-[2-(2-morpholin-4-ylethylamino)phenyl]methanone / [6-Hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophen-3-yl]{2-[(2-morpholinoethyl)amino]phenyl}methanone


Mass: 474.571 Da / Num. of mol.: 4 / Source method: obtained synthetically / Formula: C27H26N2O4S / Feature type: SUBJECT OF INVESTIGATION
#5: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 526 / Source method: isolated from a natural source / Formula: H2O
Has ligand of interestY
Has protein modificationN

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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: 2.38 Å3/Da / Density % sol: 48.4 %
Crystal growTemperature: 293 K / Method: vapor diffusion, sitting drop
Details: 18.3% (w/v) PEG 3350, 0.1 M Bis-TRIS pH 6.56, 0.133 M lithium acetate

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Data collection

DiffractionMean temperature: 100 K / Serial crystal experiment: N
Diffraction sourceSource: SYNCHROTRON / Site: PETRA III, DESY / Beamline: P11 / Wavelength: 1.0332 Å
DetectorType: DECTRIS EIGER2 X 16M / Detector: PIXEL / Date: Apr 22, 2022
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 1.0332 Å / Relative weight: 1
ReflectionResolution: 1.57→68.83 Å / Num. obs: 109154 / % possible obs: 99 % / Redundancy: 7 % / CC1/2: 0.999 / Rmerge(I) obs: 0.059 / Rpim(I) all: 0.024 / Rrim(I) all: 0.063 / Net I/σ(I): 14
Reflection shellResolution: 1.57→1.59 Å / Redundancy: 7 % / Rmerge(I) obs: 0.634 / Mean I/σ(I) obs: 2.4 / Num. unique obs: 5405 / CC1/2: 0.938 / Rpim(I) all: 0.254 / Rrim(I) all: 0.684 / % possible all: 98.9

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Processing

Software
NameVersionClassification
XDSJan 10, 2022data reduction
Aimlessv0.7.7data scaling
PHASERv2.8.3phasing
PHENIXv1.20.1-4487refinement
autoPROCv1.0.5 (20211020)data processing
PDB_EXTRACTv4.2data extraction
RefinementMethod to determine structure: MOLECULAR REPLACEMENT / Resolution: 1.57→53.07 Å / SU ML: 0.16 / Cross valid method: THROUGHOUT / σ(F): 1.35 / Phase error: 22.56 / Stereochemistry target values: ML
RfactorNum. reflection% reflection
Rfree0.1827 5505 5.06 %
Rwork0.1637 --
obs0.1647 108813 98.69 %
Solvent computationShrinkage radii: 0.9 Å / VDW probe radii: 1.1 Å / Solvent model: FLAT BULK SOLVENT MODEL
Displacement parametersBiso mean: 27.86 Å2
Refinement stepCycle: LAST / Resolution: 1.57→53.07 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms5057 0 188 526 5771
Refine LS restraints
Refine-IDTypeDev idealNumber
X-RAY DIFFRACTIONf_bond_d0.0085590
X-RAY DIFFRACTIONf_angle_d0.7917590
X-RAY DIFFRACTIONf_dihedral_angle_d17.7372050
X-RAY DIFFRACTIONf_chiral_restr0.061710
X-RAY DIFFRACTIONf_plane_restr0.0081018
LS refinement shell
Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkRefine-ID% reflection obs (%)
1.57-1.580.31061910.28193390X-RAY DIFFRACTION98
1.58-1.60.25861900.24413451X-RAY DIFFRACTION99
1.6-1.620.26581780.23293425X-RAY DIFFRACTION99
1.62-1.640.21341520.21353451X-RAY DIFFRACTION99
1.64-1.670.23922210.20413427X-RAY DIFFRACTION99
1.67-1.690.20561680.19363465X-RAY DIFFRACTION99
1.69-1.710.22961900.18483418X-RAY DIFFRACTION99
1.71-1.740.20741800.17583437X-RAY DIFFRACTION99
1.74-1.760.23151690.17713495X-RAY DIFFRACTION99
1.76-1.790.18871680.17453410X-RAY DIFFRACTION99
1.79-1.820.19111900.17983443X-RAY DIFFRACTION99
1.82-1.860.20271890.17853467X-RAY DIFFRACTION99
1.86-1.890.20931740.17983465X-RAY DIFFRACTION99
1.89-1.930.21361900.17583419X-RAY DIFFRACTION99
1.93-1.970.21162070.1623412X-RAY DIFFRACTION99
1.97-2.020.18992080.16063397X-RAY DIFFRACTION99
2.02-2.070.17731610.15453437X-RAY DIFFRACTION97
2.07-2.130.18952130.15763266X-RAY DIFFRACTION95
2.13-2.190.17981780.15383421X-RAY DIFFRACTION98
2.19-2.260.17061770.15343474X-RAY DIFFRACTION99
2.26-2.340.18581950.15613441X-RAY DIFFRACTION99
2.34-2.430.16661860.15123442X-RAY DIFFRACTION99
2.43-2.550.1781810.15213468X-RAY DIFFRACTION99
2.55-2.680.18681360.15823566X-RAY DIFFRACTION99
2.68-2.850.16212280.15213425X-RAY DIFFRACTION99
2.85-3.070.15221540.15663504X-RAY DIFFRACTION99
3.07-3.380.19191930.16083504X-RAY DIFFRACTION99
3.38-3.860.17791940.14433415X-RAY DIFFRACTION97
3.86-4.870.12881970.13173415X-RAY DIFFRACTION97
4.87-53.070.20451470.20353558X-RAY DIFFRACTION97
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)
16.837-3.47981.43115.2694-13.08-0.0746-0.17820.10330.18520.0239-0.4169-0.12731.12730.11990.2251-0.04280.01790.5569-0.01060.174-17.980419.5246-12.6707
24.7062-3.8085-3.30153.59613.42323.46550.25050.2640.0998-0.2865-0.20880.0835-0.5841-0.0563-0.03220.3008-0.00390.0110.3210.01760.1882-31.580823.6245-22.9871
33.1215-0.0064-5.29571.2507-0.10859.2222-0.33150.2568-0.5373-0.07090.0329-0.04570.5988-0.10620.33370.1973-0.01390.00790.2315-0.04570.1709-37.054310.0324-13.8224
43.6432-4.8763-2.57598.01261.17135.44790.09910.3761-0.0521-0.5649-0.08480.1076-0.0604-0.25610.05840.2228-0.0268-0.01350.3286-0.03430.1612-38.613217.4922-23.3333
54.2063-1.59810.97341.7871-0.81993.04530.0330.11610.1725-0.083-0.027-0.0698-0.22620.23390.00520.1662-0.02790.01070.1593-0.00460.1011-30.180522.8524-8.0332
64.9811-0.6438-1.08071.0422-1.05795.1912-0.00230.70290.4101-0.30180.09390.16340.0319-1.0824-0.06390.24670.0595-0.03980.4470.03570.2057-51.996523.7266-16.1108
71.59960.0324-0.60050.6384-0.15292.7840.01820.0670.03970.04750.01980.0409-0.0928-0.2546-0.070.1562-0.0017-0.0030.1752-0.00580.1132-41.338119.034-3.9716
89.05224.45670.11715.9011-1.15593.1875-0.0848-0.0702-0.5019-0.1474-0.0248-0.49840.09720.85510.10880.19180.0505-0.01580.4614-0.00220.1731-17.708813.511910.6165
94.61852.51982.25756.7493.80014.32710.0486-0.8391-0.60240.3114-0.1057-0.010.35470.0273-0.01640.2140.03180.00560.46260.14010.2138-31.05569.728320.7869
103.94881.28785.17031.69061.47457.152-0.2398-0.37680.52610.0295-0.02490.0301-0.4913-0.04250.32250.19410.0146-0.00840.2665-0.05380.1615-36.390823.101112.7267
114.57795.06544.59695.94064.64275.1917-0.1206-0.31720.15590.293-0.03160.2968-0.14470.05550.12870.24180.0269-0.00150.3870.0010.1834-38.307915.585521.4291
123.57941.3025-1.40261.6528-0.83682.1822-0.0967-0.1448-0.19330.03070.0061-0.08850.1370.23960.10450.15790.031-0.010.17260.01450.1126-30.20999.99585.922
134.58971.0957-0.25581.03460.14431.7625-0.0316-0.2243-0.14910.0432-0.0129-0.00860.020.04910.03590.1550.02360.0060.15050.00840.0974-38.947513.26068.2248
140.671-0.76182.25731.5538-3.31798.3888-0.01050.51810.0307-0.19770.16480.1010.3412-0.8306-0.01550.2149-0.0472-0.02270.4831-0.01620.2193-52.699712.6844-11.113
156.7036-4.5457-0.41096.1054-0.83183.8932-0.1043-0.40420.32070.08910.0625-0.4424-0.07020.72920.09010.1899-0.05720.010.3719-0.01250.1612-51.808619.2833-53.9932
165.9853-4.4758-3.39924.21823.78543.72340.13310.45560.1369-0.2041-0.20380.1765-0.2741-0.10650.10120.2392-0.0241-0.00060.32840.02420.1731-65.349723.6204-64.1552
174.665-1.4231-4.6992.0951.32674.7793-0.30390.4167-0.5747-0.0681-0.01710.06580.3762-0.1820.34620.1831-0.02910.00610.264-0.05350.1621-71.338510.203-55.841
184.2779-4.8038-2.47655.6331.90885.07020.0290.5825-0.2752-0.4452-0.15570.39450.2057-0.27610.10990.2537-0.0181-0.02310.3635-0.0230.1963-72.921817.9108-65.036
193.8729-1.38291.05661.8652-0.98122.6525-0.05340.10890.2122-0.01960.0198-0.0533-0.16660.17670.04370.1609-0.03020.01010.17250.01450.1165-64.10622.5668-49.4168
204.4759-1.2161-0.79081.04790.21272.1045-0.04090.19580.07180.00640.0150.0264-0.0272-0.13840.02340.1775-0.0205-0.00770.18650.00210.1175-72.984619.6917-51.5717
210.51270.7303-1.61322.8889-4.89338.70940.0069-0.22020.0240.25850.16440.1264-0.3203-1.1343-0.15820.18720.05220.03330.3388-0.00430.1821-86.89919.843-32.1246
223.41913.0353-1.57943.541-1.07732.5147-0.2267-0.1203-0.3306-0.12650.0161-0.3820.17121.17920.11370.21610.0566-0.01830.59220.00860.1953-52.763712.665-30.855
233.60063.3349-0.44213.1978-0.33950.14620.0692-0.6904-0.31520.0735-0.1874-0.15470.09340.2840.07340.23920.030500.41710.07250.1834-65.90268.8009-20.7863
243.60780.28154.68712.2967-0.28056.7402-0.3206-0.31380.56550.11290.03160.0227-0.5632-0.02970.34760.2120.01580.00030.2538-0.04890.1809-71.097222.5815-29.3754
254.62795.57694.17237.04084.08886.20320.0652-0.39860.14140.3885-0.13160.3811-0.16720.18870.08320.26450.02730.0150.3682-0.01210.2273-73.446614.7578-20.1455
264.37111.3209-1.5311.461-0.57852.2262-0.0927-0.1762-0.30220.047-0.0282-0.07390.1680.30110.12460.15980.0266-0.00310.18090.02070.1169-64.79279.6752-35.8418
276.20791.7126-0.51711.19180.33752.399-0.062-0.1021-0.30750.0495-0.01110.00120.10620.01170.07280.17140.03590.00550.12420.03340.132-74.092111.0187-32.7471
283.15680.6451-1.27960.6667-0.65310.8356-0.083-0.0526-0.0140.02410.04890.0008-0.0373-0.00120.04610.15940.01210.00330.1688-0.0140.108-72.234615.4435-34.6226
290.3604-0.44031.16141.6248-2.80765.5329-0.01080.6035-0.0333-0.35530.03010.10080.5515-0.9254-0.03850.2551-0.023-0.03190.5466-0.02460.2172-86.901813.8628-53.1573
Refinement TLS group
IDRefine-IDRefine TLS-IDSelection details
1X-RAY DIFFRACTION1chain 'A' and (resid 12 through 32 )
2X-RAY DIFFRACTION2chain 'A' and (resid 33 through 44 )
3X-RAY DIFFRACTION3chain 'A' and (resid 45 through 66 )
4X-RAY DIFFRACTION4chain 'A' and (resid 67 through 80 )
5X-RAY DIFFRACTION5chain 'A' and (resid 81 through 110 )
6X-RAY DIFFRACTION6chain 'A' and (resid 111 through 117 )
7X-RAY DIFFRACTION7chain 'A' and (resid 118 through 165 )
8X-RAY DIFFRACTION8chain 'B' and (resid 12 through 32 )
9X-RAY DIFFRACTION9chain 'B' and (resid 33 through 44 )
10X-RAY DIFFRACTION10chain 'B' and (resid 45 through 66 )
11X-RAY DIFFRACTION11chain 'B' and (resid 67 through 80 )
12X-RAY DIFFRACTION12chain 'B' and (resid 81 through 110 )
13X-RAY DIFFRACTION13chain 'B' and (resid 111 through 151 )
14X-RAY DIFFRACTION14chain 'B' and (resid 152 through 165 )
15X-RAY DIFFRACTION15chain 'C' and (resid 12 through 32 )
16X-RAY DIFFRACTION16chain 'C' and (resid 33 through 44 )
17X-RAY DIFFRACTION17chain 'C' and (resid 45 through 66 )
18X-RAY DIFFRACTION18chain 'C' and (resid 67 through 80 )
19X-RAY DIFFRACTION19chain 'C' and (resid 81 through 110 )
20X-RAY DIFFRACTION20chain 'C' and (resid 111 through 151 )
21X-RAY DIFFRACTION21chain 'C' and (resid 152 through 165 )
22X-RAY DIFFRACTION22chain 'D' and (resid 13 through 32 )
23X-RAY DIFFRACTION23chain 'D' and (resid 33 through 44 )
24X-RAY DIFFRACTION24chain 'D' and (resid 45 through 66 )
25X-RAY DIFFRACTION25chain 'D' and (resid 67 through 80 )
26X-RAY DIFFRACTION26chain 'D' and (resid 81 through 110 )
27X-RAY DIFFRACTION27chain 'D' and (resid 111 through 131 )
28X-RAY DIFFRACTION28chain 'D' and (resid 132 through 151 )
29X-RAY DIFFRACTION29chain 'D' and (resid 152 through 165 )

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