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- PDB-7t31: X-ray Structure of Clostridiodies difficile PilW -

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Basic information

Entry
Database: PDB / ID: 7t31
TitleX-ray Structure of Clostridiodies difficile PilW
ComponentsMaltose/maltodextrin-binding periplasmic protein,Putative pilin protein chimera
KeywordsDNA BINDING PROTEIN / type IV pilin / gram-positive
Function / homology
Function and homology information


protein secretion by the type II secretion system / type II protein secretion system complex / detection of maltose stimulus / maltose binding / maltose transport complex / maltose transport / maltodextrin transmembrane transport / carbohydrate transmembrane transporter activity / ATP-binding cassette (ABC) transporter complex, substrate-binding subunit-containing / carbohydrate transport ...protein secretion by the type II secretion system / type II protein secretion system complex / detection of maltose stimulus / maltose binding / maltose transport complex / maltose transport / maltodextrin transmembrane transport / carbohydrate transmembrane transporter activity / ATP-binding cassette (ABC) transporter complex, substrate-binding subunit-containing / carbohydrate transport / ATP-binding cassette (ABC) transporter complex / cell chemotaxis / outer membrane-bounded periplasmic space / membrane => GO:0016020 / periplasmic space / DNA damage response / membrane
Similarity search - Function
Bacterial general secretion pathway protein G-type pilin / Prokaryotic N-terminal methylation site. / Prokaryotic N-terminal methylation motif / Prokaryotic N-terminal methylation site / Pilin-like / Maltose/Cyclodextrin ABC transporter, substrate-binding protein / Solute-binding family 1, conserved site / Bacterial extracellular solute-binding proteins, family 1 signature. / Bacterial extracellular solute-binding protein / Bacterial extracellular solute-binding protein
Similarity search - Domain/homology
Maltose/maltodextrin-binding periplasmic protein / Putative pilin protein
Similarity search - Component
Biological speciesClostridioides difficile R20291 (bacteria)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / molecular replacement / Resolution: 2.3 Å
AuthorsRonish, L.A. / Piepenbrink, K.H.
Funding support United States, 2items
OrganizationGrant numberCountry
National Institutes of Health/National Institute Of Allergy and Infectious Diseases (NIH/NIAID)K22 AI123467 United States
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)P20-GM113126 United States
CitationJournal: J.Biol.Chem. / Year: 2022
Title: Recognition of extracellular DNA by type IV pili promotes biofilm formation by Clostridioides difficile.
Authors: Ronish, L.A. / Sidner, B. / Yu, Y. / Piepenbrink, K.H.
History
DepositionDec 6, 2021Deposition site: RCSB / Processing site: RCSB
Revision 1.0Oct 19, 2022Provider: repository / Type: Initial release
Revision 1.1Oct 26, 2022Group: Database references / Category: citation / Item: _citation.journal_volume
Revision 1.2Apr 3, 2024Group: Data collection / Refinement description
Category: chem_comp_atom / chem_comp_bond / pdbx_initial_refinement_model

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

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Assembly

Deposited unit
A: Maltose/maltodextrin-binding periplasmic protein,Putative pilin protein chimera
B: Maltose/maltodextrin-binding periplasmic protein,Putative pilin protein chimera
C: Maltose/maltodextrin-binding periplasmic protein,Putative pilin protein chimera
D: Maltose/maltodextrin-binding periplasmic protein,Putative pilin protein chimera


Theoretical massNumber of molelcules
Total (without water)223,9574
Polymers223,9574
Non-polymers00
Water17,925995
1
A: Maltose/maltodextrin-binding periplasmic protein,Putative pilin protein chimera


Theoretical massNumber of molelcules
Total (without water)55,9891
Polymers55,9891
Non-polymers00
Water181
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
2
B: Maltose/maltodextrin-binding periplasmic protein,Putative pilin protein chimera


Theoretical massNumber of molelcules
Total (without water)55,9891
Polymers55,9891
Non-polymers00
Water181
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
3
C: Maltose/maltodextrin-binding periplasmic protein,Putative pilin protein chimera


Theoretical massNumber of molelcules
Total (without water)55,9891
Polymers55,9891
Non-polymers00
Water181
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
4
D: Maltose/maltodextrin-binding periplasmic protein,Putative pilin protein chimera


Theoretical massNumber of molelcules
Total (without water)55,9891
Polymers55,9891
Non-polymers00
Water181
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Unit cell
Length a, b, c (Å)65.244, 81.754, 102.797
Angle α, β, γ (deg.)92.31, 91.05, 113.37
Int Tables number1
Space group name H-MP1

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Components

#1: Protein
Maltose/maltodextrin-binding periplasmic protein,Putative pilin protein chimera / MMBP / Maltodextrin-binding protein / Maltose-binding protein / MBP


Mass: 55989.332 Da / Num. of mol.: 4
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Clostridioides difficile R20291 (bacteria)
Strain: K12, 630 / Gene: malE, b4034, JW3994, CD630_23050 / Plasmid: petMal / Production host: Escherichia coli (E. coli) / Strain (production host): NiCO21 / References: UniProt: P0AEX9, UniProt: Q185H8
#2: Water ChemComp-HOH / water / Water


Mass: 18.015 Da / Num. of mol.: 995 / 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: 2.35 Å3/Da / Density % sol: 47.57 % / Description: Rods
Crystal growTemperature: 280 K / Method: vapor diffusion, hanging drop / pH: 7.5
Details: 0.1M Hepes pH 7.5, 25% Peg 3350, 0.0375M NaCl, 0.2M CsCl2, 2% ethanol

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Data collection

DiffractionMean temperature: 100 K / Serial crystal experiment: N
Diffraction sourceSource: SYNCHROTRON / Site: APS / Beamline: 23-ID-D / Wavelength: 1.0332 Å
DetectorType: DECTRIS PILATUS3 6M / Detector: PIXEL / Date: Oct 30, 2020
RadiationMonochromator: double crystal / Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 1.0332 Å / Relative weight: 1
ReflectionResolution: 2.01→74.95 Å / Num. obs: 121248 / % possible obs: 93.1 % / Redundancy: 1.8 % / Biso Wilson estimate: 28.39 Å2 / CC1/2: 0.995 / Rmerge(I) obs: 0.074 / Rpim(I) all: 0.074 / Rrim(I) all: 0.104 / Net I/σ(I): 3.1 / Num. measured all: 216621
Reflection shell

Diffraction-ID: 1 / Redundancy: 1.8 %

Resolution (Å)Rmerge(I) obsNum. measured allNum. unique obsCC1/2Rpim(I) allRrim(I) allNet I/σ(I) obs% possible all
2.01-2.121.10633001180850.3951.1051.5630.595.1
6.35-74.950.031682538680.9950.0310.0448.794

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Phasing

PhasingMethod: molecular replacement
Phasing MR
Highest resolutionLowest resolution
Rotation3 Å102.64 Å

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Processing

Software
NameVersionClassificationNB
PHENIX(1.19.2_4158: ???)refinement
XDSdata reduction
Aimless0.7.4data scaling
PHASER2.8.0phasing
PDB_EXTRACT3.27data extraction
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: MBP, PilA1

Resolution: 2.3→39.7 Å / SU ML: 0.3 / Cross valid method: THROUGHOUT / σ(F): 1.96 / Phase error: 25.86 / Stereochemistry target values: ML
RfactorNum. reflection% reflection
Rfree0.2453 3522 4.98 %
Rwork0.1961 --
obs0.1985 70741 81.73 %
Solvent computationShrinkage radii: 0.9 Å / VDW probe radii: 1.11 Å / Solvent model: FLAT BULK SOLVENT MODEL
Refinement stepCycle: LAST / Resolution: 2.3→39.7 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms14998 0 0 995 15993
Refine LS restraints
Refine-IDTypeDev idealNumber
X-RAY DIFFRACTIONf_bond_d0.00215413
X-RAY DIFFRACTIONf_angle_d0.53920928
X-RAY DIFFRACTIONf_dihedral_angle_d18.2145642
X-RAY DIFFRACTIONf_chiral_restr0.0422315
X-RAY DIFFRACTIONf_plane_restr0.0032702
LS refinement shell
Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkRefine-ID% reflection obs (%)
2.3-2.330.3377790.25941429X-RAY DIFFRACTION42
2.33-2.360.3218700.24831576X-RAY DIFFRACTION49
2.36-2.40.3047690.25081839X-RAY DIFFRACTION55
2.4-2.440.2851820.23242018X-RAY DIFFRACTION62
2.44-2.480.28881220.24372168X-RAY DIFFRACTION66
2.48-2.520.26921290.24412362X-RAY DIFFRACTION72
2.52-2.570.29331650.25642521X-RAY DIFFRACTION77
2.57-2.620.3071480.25962768X-RAY DIFFRACTION84
2.62-2.670.26461380.24032868X-RAY DIFFRACTION88
2.67-2.730.30911670.23353103X-RAY DIFFRACTION93
2.73-2.790.3081580.23243064X-RAY DIFFRACTION94
2.79-2.860.29621750.22663082X-RAY DIFFRACTION94
2.86-2.940.28961370.23073058X-RAY DIFFRACTION92
2.94-3.020.2851350.22753015X-RAY DIFFRACTION92
3.02-3.120.27771690.23043011X-RAY DIFFRACTION92
3.12-3.230.26031690.21852945X-RAY DIFFRACTION90
3.23-3.360.2641700.20042954X-RAY DIFFRACTION91
3.36-3.510.24681410.18142897X-RAY DIFFRACTION87
3.52-3.70.22491360.17182863X-RAY DIFFRACTION87
3.7-3.930.21041580.17242852X-RAY DIFFRACTION87
3.93-4.230.19031790.15712804X-RAY DIFFRACTION86
4.24-4.660.19761500.1462961X-RAY DIFFRACTION90
4.66-5.330.19311530.15622964X-RAY DIFFRACTION90
5.33-6.710.23911590.18683010X-RAY DIFFRACTION92
6.72-39.70.21291640.17023087X-RAY DIFFRACTION94
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)
16.87963.18491.16656.80792.7085.514-0.75211.50790.4577-1.9220.42060.2984-0.74620.10060.23780.8144-0.1758-0.08570.93390.08520.44221.0143-70.580923.2892
25.62851.00950.62460.6895-0.05361.816-0.108-0.08580.58410.18710.133-0.0156-0.0171-0.3523-0.0130.32640.0609-0.04530.2632-0.05230.341733.3775-72.860338.3738
37.0013-1.4152-1.15484.04411.00842.9090.20570.23950.18860-0.07960.1581-0.20880.0034-0.12960.21750.0599-0.00170.2192-0.06680.386154.8014-71.259138.8088
42.8036-0.3975-0.470.2243-0.35811.01980.13060.0773-0.25920.01070.1162-0.0854-0.1992-0.0633-0.2090.27660.10260.05220.3068-0.09010.475636.1211-68.412944.5347
57.08051.2455-2.01644.8901-0.81537.88670.9854-1.94150.59080.6723-0.1819-0.1963-1.54640.3328-0.62220.90510.06490.05891.1573-0.2010.6128.392-46.770682.6554
64.537-4.0524-1.43635.37342.85355.80070.3122-1.56121.43550.47520.4690.1328-0.95990.5153-0.63831.04010.20790.37410.7685-0.17481.065431.7262-41.100173.4472
75.8784-3.8321-1.28975.21630.21378.51960.2376-0.09332.8091-0.163-0.137-0.8823-1.53630.4834-0.24491.22320.05640.39860.5296-0.04191.492333.2842-38.135563.7932
86.4309-0.01010.83470.72830.42751.28110.0314-1.0035-0.89380.19180.0911-0.28380.11330.1015-0.07820.22320.0292-0.05240.32690.08530.356347.086-38.137236.2362
94.44080.11380.40650.69590.24660.73090.04-0.0546-0.24930.0646-0.05910.03660.0695-0.02880.01110.1684-0.0109-0.01180.14280.00720.090725.3259-32.505833.427
100.34280.14750.42041.10860.54481.19290.1224-0.2217-0.05880.4287-0.1283-0.28080.0717-0.2004-0.03190.3314-0.0443-0.1060.4994-0.03220.229932.975-22.450554.0914
116.53213.5685-0.01813.38780.19337.04470.16910.38182.6631-0.03260.4394-0.155-1.5295-0.3916-0.04140.9099-0.0616-0.06440.7033-0.07451.393945.2091-0.622571.2113
126.88321.45771.75064.2311-0.26645.23020.4551-1.47660.44810.9689-0.36010.1662-0.0233-0.5971-0.05541.0297-0.3547-0.41571.28730.2040.503239.707-13.949178.3535
135.34460.0995-0.44450.49630.35540.93030.0098-0.2782-0.65110.0987-0.08290.00360.1704-0.19760.04050.2856-0.06640.04460.21460.03110.26528.5334-63.2652105.2464
145.61470.79171.32744.00021.05813.86810.20480.56990.016-0.2520.01530.20070.252-0.0531-0.19070.23420.02980.0140.1808-0.00880.137653.0863-65.492293.0604
154.24340.1712-0.20.13870.15550.53470.1781-0.46620.45840.0308-0.10150.0321-0.0179-0.0765-0.05770.2039-0.05350.04410.1761-0.0530.284640.6694-58.4759106.6958
162.22130.930.68760.35590.14650.72260.14950.4266-0.1638-0.14020.1447-0.08010.17340.0218-0.28250.31690.0074-0.08450.2862-0.0630.343639.354-76.080885.0623
177.58351.11482.58484.44471.90148.0195-0.22391.5459-0.2773-1.23480.04170.28890.04410.11430.06080.9161-0.1825-0.29941.0533-0.10280.612626.3032-90.640258.4466
186.48853.6797-0.59627.24390.71353.15770.18420.1637-2.12080.41590.044-0.81660.53470.523-0.38740.7958-0.0699-0.41190.7993-0.04330.80533.8358-96.401873.613
194.4295-0.7931-0.93262.5182-0.53263.06610.41490.07141.4179-0.05610.0444-0.4-0.93180.0306-0.10840.46940.01480.13910.1978-0.02160.572884.1731-13.156104.9894
203.76122.00482.40284.1654-0.74465.5215-0.3441.50210.4889-0.66030.2687-0.0188-0.2943-0.28590.11310.43550.11930.05820.60660.1660.231978.8555-20.528893.1133
215.23850.5797-1.10294.201-2.86183.4538-0.18720.5887-0.7889-0.26470.1017-0.0840.7391-0.1708-0.00230.2435-0.0138-0.01690.1771-0.0490.261179.8247-33.8922102.7071
221.76650.20391.02030.67880.22080.59920.02370.35190.0312-0.01660.01420.04950.03290.0904-0.03640.2559-0.00070.02110.21220.04220.102951.5186-32.2406100.6593
234.3870.117-0.5530.5956-0.13080.0733-0.0255-0.16790.3110.00690.0491-0.0226-0.05860.09020.00020.26480.00840.020.1716-0.00060.099462.96-23.5279108.088
240.8041-0.4789-0.76931.27520.06960.82290.19090.3403-0.1409-0.5531-0.2253-0.2541-0.1036-0.0784-0.03010.44240.12060.11630.6775-0.04660.148165.4439-38.047184.1175
257.0063-4.9509-1.54657.73292.61920.96460.08431.4901-1.6899-0.129-0.84640.26680.1585-0.570.83670.9650.02910.15630.9168-0.0880.728476.8942-57.809464.5223
262.2817-0.9777-2.38552.362.97874.47070.30331.24250.5043-0.6906-0.34390.2772-0.8939-1.4637-0.14221.23030.4180.34561.41130.22930.528370.3501-43.63861.261
Refinement TLS group
IDRefine-IDRefine TLS-IDSelection details
1X-RAY DIFFRACTION1chain 'A' and (resid 1 through 42 )
2X-RAY DIFFRACTION2chain 'A' and (resid 43 through 131 )
3X-RAY DIFFRACTION3chain 'A' and (resid 132 through 245 )
4X-RAY DIFFRACTION4chain 'A' and (resid 246 through 1053 )
5X-RAY DIFFRACTION5chain 'A' and (resid 1054 through 1091 )
6X-RAY DIFFRACTION6chain 'A' and (resid 1092 through 1121 )
7X-RAY DIFFRACTION7chain 'A' and (resid 1122 through 1152 )
8X-RAY DIFFRACTION8chain 'B' and (resid 0 through 118 )
9X-RAY DIFFRACTION9chain 'B' and (resid 119 through 314 )
10X-RAY DIFFRACTION10chain 'B' and (resid 315 through 1054 )
11X-RAY DIFFRACTION11chain 'B' and (resid 1055 through 1095 )
12X-RAY DIFFRACTION12chain 'B' and (resid 1096 through 1151 )
13X-RAY DIFFRACTION13chain 'C' and (resid 1 through 131 )
14X-RAY DIFFRACTION14chain 'C' and (resid 132 through 200 )
15X-RAY DIFFRACTION15chain 'C' and (resid 201 through 314 )
16X-RAY DIFFRACTION16chain 'C' and (resid 315 through 1053 )
17X-RAY DIFFRACTION17chain 'C' and (resid 1054 through 1110 )
18X-RAY DIFFRACTION18chain 'C' and (resid 1111 through 1152 )
19X-RAY DIFFRACTION19chain 'D' and (resid 0 through 42 )
20X-RAY DIFFRACTION20chain 'D' and (resid 43 through 72 )
21X-RAY DIFFRACTION21chain 'D' and (resid 73 through 105 )
22X-RAY DIFFRACTION22chain 'D' and (resid 106 through 200 )
23X-RAY DIFFRACTION23chain 'D' and (resid 201 through 314 )
24X-RAY DIFFRACTION24chain 'D' and (resid 315 through 1054 )
25X-RAY DIFFRACTION25chain 'D' and (resid 1055 through 1110 )
26X-RAY DIFFRACTION26chain 'D' and (resid 1111 through 1152 )

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