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- PDB-8ih8: anti-sigmaF factor and Anti-sigmaF factor antagonist complex(usfx... -

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Basic information

Entry
Database: PDB / ID: 8ih8
Titleanti-sigmaF factor and Anti-sigmaF factor antagonist complex(usfx-RsfB)
Components
  • Anti-sigma-F factor RsbW
  • Anti-sigma-F factor antagonist RsfB
KeywordsTRANSCRIPTION / tetramer / complex
Function / homology
Function and homology information


anti-sigma factor antagonist activity / sigma factor antagonist activity / hydrolase activity / nucleotide binding / negative regulation of DNA-templated transcription / regulation of DNA-templated transcription / ATP binding
Similarity search - Function
Anti-sigma factor antagonist / STAS domain / STAS domain profile. / STAS domain / STAS domain superfamily
Similarity search - Domain/homology
Anti-sigma-F factor antagonist RsfB / Anti-sigma-F factor RsbW
Similarity search - Component
Biological speciesMycobacterium tuberculosis (bacteria)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 2 Å
AuthorsChen, Y.J. / Su, D.
Funding support China, 1items
OrganizationGrant numberCountry
National Natural Science Foundation of China (NSFC)31370735 China
CitationJournal: To Be Published
Title: High resolutions crystal structure of anti-sigmaF factor and Anti-sigmaF factor antagonist complex(usfx-RsfB)
Authors: Chen, Y.J. / Su, D.
History
DepositionFeb 22, 2023Deposition site: PDBJ / Processing site: PDBC
Revision 1.0Feb 28, 2024Provider: repository / Type: Initial release

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: Anti-sigma-F factor antagonist RsfB
C: Anti-sigma-F factor RsbW
B: Anti-sigma-F factor antagonist RsfB
D: Anti-sigma-F factor RsbW
hetero molecules


Theoretical massNumber of molelcules
Total (without water)56,8495
Polymers56,7564
Non-polymers921
Water4,810267
1


  • Idetical with deposited unit
  • defined by author&software
  • Evidence: gel filtration
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area5510 Å2
ΔGint-36 kcal/mol
Surface area21420 Å2
MethodPISA
Unit cell
Length a, b, c (Å)50.425, 77.928, 133.530
Angle α, β, γ (deg.)90.00, 90.00, 90.00
Int Tables number19
Space group name H-MP212121

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Components

#1: Protein Anti-sigma-F factor antagonist RsfB / Anti-anti-sigma F factor RsfB


Mass: 12808.670 Da / Num. of mol.: 2
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv) (bacteria)
Gene: rsfB, Rv3687c / Production host: Escherichia coli (E. coli) / References: UniProt: P9WGE1
#2: Protein Anti-sigma-F factor RsbW / Anti-sigma-F factor UsfX / Regulator of SigF / Sigma-F anti-sigma factor RsbW


Mass: 15569.563 Da / Num. of mol.: 2
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv) (bacteria)
Gene: rsbW, usfX, Rv3287c / Production host: Escherichia coli (E. coli) / References: UniProt: P9WGX7
#3: Chemical ChemComp-GOL / GLYCEROL / GLYCERIN / PROPANE-1,2,3-TRIOL


Mass: 92.094 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: C3H8O3
#4: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 267 / Source method: isolated from a natural source / Formula: H2O
Has ligand of interestN

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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: 2.32 Å3/Da / Density % sol: 46.92 %
Crystal growTemperature: 293 K / Method: vapor diffusion, hanging drop
Details: 0.05M HEPES pH7.5 0.2M KCl 35.5% v/v pentaerythritol propoxylate (5/4 PO/OH)

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Data collection

DiffractionMean temperature: 80 K / Serial crystal experiment: N
Diffraction sourceSource: SYNCHROTRON / Site: SSRF / Beamline: BL19U1 / Wavelength: 0.9785 Å
DetectorType: DECTRIS PILATUS3 6M / Detector: PIXEL / Date: Dec 2, 2020
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.9785 Å / Relative weight: 1
ReflectionResolution: 2→50 Å / Num. obs: 36351 / % possible obs: 99.3 % / Redundancy: 12.7 % / Rmerge(I) obs: 0.057 / Net I/σ(I): 42.7
Reflection shellResolution: 2→2.06 Å / Num. unique obs: 21537 / CC1/2: 0.15

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Processing

Software
NameVersionClassification
PHENIX(1.18.2_3874: ???)refinement
HKL-3000data reduction
HKL-3000data scaling
PHASERphasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT / Resolution: 2→33.65 Å / SU ML: 0.19 / Cross valid method: THROUGHOUT / σ(F): 1.36 / Phase error: 21.68 / Stereochemistry target values: ML
RfactorNum. reflection% reflection
Rfree0.215 2000 5.51 %
Rwork0.1804 --
obs0.1824 36275 99.55 %
Solvent computationShrinkage radii: 0.9 Å / VDW probe radii: 1.11 Å / Solvent model: FLAT BULK SOLVENT MODEL
Refinement stepCycle: LAST / Resolution: 2→33.65 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms3645 0 6 267 3918
Refine LS restraints
Refine-IDTypeDev idealNumber
X-RAY DIFFRACTIONf_bond_d0.013733
X-RAY DIFFRACTIONf_angle_d1.395088
X-RAY DIFFRACTIONf_dihedral_angle_d14.454534
X-RAY DIFFRACTIONf_chiral_restr0.067642
X-RAY DIFFRACTIONf_plane_restr0.011660
LS refinement shell
Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkRefine-ID% reflection obs (%)
2-2.050.27231360.19862325X-RAY DIFFRACTION97
2.05-2.10.25231420.19692433X-RAY DIFFRACTION100
2.1-2.170.22731400.18352404X-RAY DIFFRACTION100
2.17-2.240.20831410.17432408X-RAY DIFFRACTION100
2.24-2.320.20031410.1792428X-RAY DIFFRACTION100
2.32-2.410.25831420.18182430X-RAY DIFFRACTION100
2.41-2.520.2381430.1922451X-RAY DIFFRACTION100
2.52-2.650.27831420.19442430X-RAY DIFFRACTION100
2.65-2.820.23061420.20272447X-RAY DIFFRACTION100
2.82-3.030.26121430.20332442X-RAY DIFFRACTION100
3.03-3.340.21241440.18852467X-RAY DIFFRACTION100
3.34-3.820.19051440.17872474X-RAY DIFFRACTION100
3.82-4.810.19191460.15262500X-RAY DIFFRACTION99
4.81-33.650.18281540.17212636X-RAY DIFFRACTION100
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)
13.55372.7263-1.55336.5623-2.43994.31590.07590.08740.35440.00050.21690.4931-0.314-0.2353-0.35590.2178-0.0220.02310.3175-0.02370.2488-4.4075-11.7376-14.9598
22.24911.2324-0.57244.3024-1.18052.5943-0.07570.17240.2091-0.38130.24380.48470.0032-0.2435-0.14070.19320.0443-0.00950.2383-0.01150.20451.4573-6.1436-15.5395
39.23934.4943-4.64424.5553-4.70024.81850.2879-0.31830.19760.3145-0.22631.1116-0.6719-0.8978-0.20260.3840.06250.01920.3408-0.07770.3227-4.3728-4.9754-7.4482
46.47431.4767-1.91356.2445-2.16042.58360.0455-0.3658-0.13630.17970.14790.1254-0.2844-0.838-0.19170.1957-0.0064-0.03890.28470.03260.2790.4784-14.6102-7.335
55.17775.2799-1.82065.5216-2.55594.0809-0.43190.5849-0.0922-0.19110.4984-0.54630.08390.1114-0.00430.2720.0256-0.01140.2008-0.00060.21268.1443-5.9917-19.9438
66.04171.1333-2.44685.4115-2.13291.54840.5552-1.19150.13821.0355-0.52230.1114-0.56940.324-0.08940.3103-0.0874-0.00840.3298-0.06570.24326.6999-4.0736-4.3659
72.80221.0226-0.70954.24671.2519.02550.4426-0.5328-0.11840.4318-0.0334-0.2945-0.5190.0649-0.15630.242-0.0577-0.02320.32680.02030.25134.5328-14.1867-1.325
83.6226-0.4066-1.18233.21860.15695.2009-0.1007-0.32580.07770.2824-0.1144-0.0937-0.0290.39480.1350.20310.00020.0040.21280.0240.244116.2716-10.2076-12.7624
94.43643.16293.75534.23216.06138.9968-0.0563-0.7347-0.47730.2387-0.1598-0.23370.37490.2336-0.00310.2113-0.0379-0.00950.44860.03580.420210.8635-16.2178-7.6934
107.2414-6.4293-2.61487.69144.41383.1939-0.9329-1.3165-1.49391.0930.4049-0.41321.00140.57650.2440.35440.00510.05780.34620.15420.59174.8265-23.8089-7.5046
113.1931-0.8587-1.08983.84440.02334.538-0.1274-0.13840.0023-0.01860.1105-0.12670.06540.2113-0.02320.1385-0.0069-0.03430.2087-0.00070.171116.34615.1853-30.2017
122.028-1.6236-0.19232.75681.60443.06670.0734-0.11110.07360.2948-0.0778-0.24520.1069-0.04890.02470.1952-0.02840.00330.2220.0210.193615.34083.482-20.058
137.319-3.3538-4.00273.5693.36638.30060.23240.2410.1754-0.229-0.0198-0.3352-0.16420.4583-0.18360.2443-0.0196-0.02340.20380.01840.201120.526511.4582-25.994
142.1985-0.72060.46275.1243-0.24262.23080.1912-0.1592-0.0229-0.1105-0.1438-0.2802-0.04640.1581-0.09520.245-0.0323-0.01220.26560.01850.187721.65249.6-12.5189
157.3355-0.9375-1.28362.40072.72686.2973-0.206-0.36180.70690.55260.2983-0.6207-0.0731-0.0218-0.13470.1993-0.0559-0.03480.27340.00520.204521.488817.2193-18.6751
164.58991.75522.62026.63015.1174.3190.1124-0.0061-0.51620.2120.538-1.1987-0.58640.5006-0.89860.50130.0410.03380.5545-0.04630.535526.2767-6.6001-42.4169
174.1318-0.2474-0.15161.8849-0.99673.29180.1797-0.1522-0.55510.2897-0.1088-0.22490.5603-0.1204-0.06940.29580.031-0.00620.2207-0.01440.231318.5156-16.9088-54.6354
183.51370.1662-0.86779.87211.97681.98880.1740.4984-0.3503-0.3734-0.036-0.63350.26540.5875-0.14940.33070.1196-0.00270.3396-0.00620.303823.7166-13.7026-61.56
193.301-1.381-0.33185.91121.82913.6803-0.04780.0552-0.40840.22930.0665-0.12370.4844-0.007-0.10670.2168-0.0068-0.01670.17260.0140.23914.1149-15.2628-56.7248
206.45351.45942.0567.36624.42172.94760.10960.7826-0.1099-1.03690.0011-0.0253-0.30650.4739-0.04520.3390.0099-0.0340.24640.03820.209614.5054-7.0624-64.0564
211.7422-0.73640.04652.59210.65472.8452-0.06560.3596-0.201-0.0897-0.21290.27140.297-0.1690.21160.2435-0.0548-0.03940.211-0.01570.20569.1289-14.9651-60.7832
227.0945-2.5350.48338.04814.49223.181-0.1961-0.0871-0.2114-0.3356-0.03470.5432-0.1519-0.87130.04910.2337-0.0221-0.01310.2763-0.02350.2732.6905-6.1209-58.3155
235.691-0.19510.08512.3244-3.3549.09970.06280.6146-0.93380.02380.03250.8690.8364-0.8371-0.08470.3946-0.1161-0.01720.3405-0.08230.46245.7733-20.9866-60.8275
243.342-3.2355-1.68233.61110.82249.3250.26020.5721.531-0.344-0.03171.0313-0.5629-1.02490.09610.43440.1866-0.04950.4739-0.02140.60853.641415.9687-33.4173
257.5433-2.7343-3.97036.90812.4382.3583-0.1753-0.11890.39580.17050.4048-0.64290.10630.5117-0.28890.1635-0.0166-0.05870.2621-0.02070.204918.921410.9999-39.3427
263.04491.13720.76583.73960.08693.67430.03190.0207-0.18460.0385-0.06670.1090.3641-0.31280.0540.2043-0.0009-0.02160.1795-0.02230.158311.35641.5502-39.9415
279.36197.7802-1.60947.7503-1.63241.9273-0.21650.19490.0781-0.33140.17480.00140.15910.0670.03250.17210.0358-0.00320.1716-0.02920.193714.15365.5188-48.6283
283.18242.7151-2.84133.8543-1.2073.88720.328-0.35830.68320.39120.02560.3668-0.1341-0.4393-0.33550.2578-0.0082-0.01080.40360.01270.25266.501512.6077-39.4511
292.8844.0782-4.05057.3833-4.11397.07690.2258-0.06250.73140.24450.19430.4913-0.0596-0.1699-0.35250.16070.0128-0.01540.2025-0.02430.17179.668713.7424-44.9
304.02114.91472.51886.76123.41151.9683-0.24640.4480.1945-0.51360.21870.309-0.09880.2521-0.02550.29540.02960.00330.35930.02090.205916.145115.203-59.1336
312.25920.09661.05843.5281.59494.06710.25870.16810.014-0.3933-0.26130.38720.2675-0.58530.03450.29110.0254-0.05360.30250.01410.2436.544210.276-55.1995
322.6565-0.7907-2.56455.20922.63893.17440.32180.70721.1749-0.4471-0.03220.1512-0.7050.3464-0.22980.34410.02830.00250.3090.02360.431322.674325.6039-53.228
338.15564.7273-2.93667.2499-2.85235.03840.2991-0.37940.31940.1604-0.20060.60290.3472-0.1932-0.05830.1986-0.03060.00420.2737-0.01710.19185.562412.3323-47.3327
Refinement TLS group
IDRefine-IDRefine TLS-IDSelection details
1X-RAY DIFFRACTION1chain 'A' and (resid 3 through 16 )
2X-RAY DIFFRACTION2chain 'A' and (resid 17 through 30 )
3X-RAY DIFFRACTION3chain 'A' and (resid 31 through 44 )
4X-RAY DIFFRACTION4chain 'A' and (resid 45 through 54 )
5X-RAY DIFFRACTION5chain 'A' and (resid 55 through 60 )
6X-RAY DIFFRACTION6chain 'A' and (resid 61 through 74 )
7X-RAY DIFFRACTION7chain 'A' and (resid 75 through 83 )
8X-RAY DIFFRACTION8chain 'A' and (resid 84 through 100 )
9X-RAY DIFFRACTION9chain 'A' and (resid 101 through 107 )
10X-RAY DIFFRACTION10chain 'A' and (resid 108 through 117 )
11X-RAY DIFFRACTION11chain 'C' and (resid 12 through 46 )
12X-RAY DIFFRACTION12chain 'C' and (resid 47 through 67 )
13X-RAY DIFFRACTION13chain 'C' and (resid 68 through 93 )
14X-RAY DIFFRACTION14chain 'C' and (resid 94 through 120 )
15X-RAY DIFFRACTION15chain 'C' and (resid 121 through 140 )
16X-RAY DIFFRACTION16chain 'B' and (resid -1 through 7 )
17X-RAY DIFFRACTION17chain 'B' and (resid 8 through 30 )
18X-RAY DIFFRACTION18chain 'B' and (resid 31 through 44 )
19X-RAY DIFFRACTION19chain 'B' and (resid 45 through 60 )
20X-RAY DIFFRACTION20chain 'B' and (resid 61 through 74 )
21X-RAY DIFFRACTION21chain 'B' and (resid 75 through 89 )
22X-RAY DIFFRACTION22chain 'B' and (resid 90 through 100 )
23X-RAY DIFFRACTION23chain 'B' and (resid 101 through 116 )
24X-RAY DIFFRACTION24chain 'D' and (resid 12 through 17 )
25X-RAY DIFFRACTION25chain 'D' and (resid 18 through 25 )
26X-RAY DIFFRACTION26chain 'D' and (resid 26 through 46 )
27X-RAY DIFFRACTION27chain 'D' and (resid 47 through 72 )
28X-RAY DIFFRACTION28chain 'D' and (resid 73 through 83 )
29X-RAY DIFFRACTION29chain 'D' and (resid 84 through 93 )
30X-RAY DIFFRACTION30chain 'D' and (resid 94 through 103 )
31X-RAY DIFFRACTION31chain 'D' and (resid 104 through 120 )
32X-RAY DIFFRACTION32chain 'D' and (resid 121 through 129 )
33X-RAY DIFFRACTION33chain 'D' and (resid 130 through 141 )

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