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- PDB-2b5a: C.BclI, Control Element of the BclI Restriction-Modification System -

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Basic information

Entry
Database: PDB / ID: 2b5a
TitleC.BclI, Control Element of the BclI Restriction-Modification System
ComponentsC.BclI
KeywordsGENE REGULATION / HELIX-TURN-HELIX motif
Function / homologylambda repressor-like DNA-binding domains / 434 Repressor (Amino-terminal Domain) / Orthogonal Bundle / Mainly Alpha / ACETIC ACID
Function and homology information
Biological speciesBacillus caldolyticus (bacteria)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MAD / Resolution: 1.543 Å
AuthorsSawaya, M.R. / Zhu, Z. / Mersha, F. / Chan, S.H. / Dabur, R. / Xu, S.Y. / Balendiran, G.K.
CitationJournal: Structure / Year: 2005
Title: Crystal Structure of the Restriction-Modification System Control Element C.BclI and Mapping of Its Binding Site.
Authors: Sawaya, M.R. / Zhu, Z. / Mersha, F. / Chan, S.H. / Dabur, R. / Xu, S.Y. / Balendiran, G.K.
History
DepositionSep 28, 2005Deposition site: RCSB / Processing site: RCSB
Revision 1.0Jan 3, 2006Provider: repository / Type: Initial release
Revision 1.1May 1, 2008Group: Version format compliance
Revision 1.2Jul 13, 2011Group: Advisory / Version format compliance
Revision 1.3Oct 11, 2017Group: Refinement description / Category: software
Item: _software.classification / _software.contact_author ..._software.classification / _software.contact_author / _software.contact_author_email / _software.date / _software.language / _software.location / _software.name / _software.type / _software.version
Revision 1.4Feb 14, 2024Group: Data collection / Database references / Derived calculations
Category: chem_comp_atom / chem_comp_bond ...chem_comp_atom / chem_comp_bond / database_2 / struct_site
Item: _database_2.pdbx_DOI / _database_2.pdbx_database_accession ..._database_2.pdbx_DOI / _database_2.pdbx_database_accession / _struct_site.pdbx_auth_asym_id / _struct_site.pdbx_auth_comp_id / _struct_site.pdbx_auth_seq_id
Remark 999SEQUENCE THERE WAS NO SUITABLE SEQUENCE DATABASE REFERENCE AT THE TIME OF PROCESSING.

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: C.BclI
B: C.BclI
C: C.BclI
D: C.BclI
hetero molecules


Theoretical massNumber of molelcules
Total (without water)35,8456
Polymers35,7254
Non-polymers1202
Water4,125229
1
A: C.BclI
B: C.BclI
hetero molecules


Theoretical massNumber of molelcules
Total (without water)17,9233
Polymers17,8632
Non-polymers601
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area1880 Å2
ΔGint-17 kcal/mol
Surface area8230 Å2
MethodPISA
2
C: C.BclI
D: C.BclI
hetero molecules


Theoretical massNumber of molelcules
Total (without water)17,9233
Polymers17,8632
Non-polymers601
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area1820 Å2
ΔGint-17 kcal/mol
Surface area8250 Å2
MethodPISA
Unit cell
Length a, b, c (Å)63.205, 65.016, 73.296
Angle α, β, γ (deg.)90.00, 90.00, 90.00
Int Tables number18
Space group name H-MP21212
DetailsThe biological assembly is a homodimer. Molecules A and B compose one biological assembly. Molecules C and D compose a second copy of the biological assembly.

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Components

#1: Protein
C.BclI


Mass: 8931.332 Da / Num. of mol.: 4
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Bacillus caldolyticus (bacteria) / Gene: bclIC / Plasmid: LITMUS28 / Production host: Escherichia coli (E. coli) / Strain (production host): ER2683
#2: Chemical ChemComp-ACY / ACETIC ACID


Mass: 60.052 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: C2H4O2
#3: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 229 / 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: 2.19 Å3/Da / Density % sol: 43.3 %
Crystal growTemperature: 298 K / Method: vapor diffusion, hanging drop / pH: 6.5
Details: 25% (w/v) Polyethylene glyco 400, 25 mM ammonium sulfate, 50 mM citrate, pH 6.5, VAPOR DIFFUSION, HANGING DROP, temperature 298K

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Data collection

Diffraction
IDMean temperature (K)Crystal-ID
11001
21
31
Diffraction source
SourceSiteBeamlineIDWavelength (Å)
SYNCHROTRONSSRL BL7-111.08
SYNCHROTRONSSRL BL1-521.0061, 1.0088, 0.9076
Detector
TypeIDDetectorDateDetails
MARRESEARCH1IMAGE PLATEMar 27, 200258 cm long, Pt-coated, fused silica, vertical focusmirror, Cyclindrically bent triangular Si(111) asymmetric cut, horizontal focus monochromator
ADSC QUANTUM 42CCDMay 24, 2002
Radiation
IDMonochromatorProtocolMonochromatic (M) / Laue (L)Scattering typeWavelength-ID
1Si (111)SINGLE WAVELENGTHMx-ray1
2MADMx-ray2
Radiation wavelength
IDWavelength (Å)Relative weight
11.081
21.00611
31.00881
40.90761
Reflection
IDNumberRmerge(I) obsΧ2D res high (Å)D res low (Å)% possible obs
1142460.0421.0691.620069.5
2140440.0391.0471.620068.5
3182120.0461.1241.620088.7
Diffraction reflection shell
Highest resolution (Å)Lowest resolution (Å)% possible obs (%)IDRmerge(I) obsChi squared
3.4520092.210.0331.042
2.743.4599.910.0351.059
2.392.7410010.0421.084
2.172.3983.510.0691.052
2.022.1797.310.0591.081
1.92.0272.310.0841.124
1.81.956.710.0831.068
1.721.845.510.0981.056
1.661.7232.610.1241.087
1.61.6610.210.1771.204
3.4520091.620.0311.113
2.743.4599.920.0321.008
2.392.7410020.0381.011
2.172.3981.520.0641.025
2.022.1796.620.0551.065
1.92.0271.620.0781.041
1.81.955.120.081.07
1.721.844.820.0991.1
1.661.7230.420.1171.168
1.61.668.520.1751.448
3.4520084.830.0331.102
2.743.4599.430.0331.062
2.392.7499.530.0381.085
2.172.3970.730.0741.093
2.022.1799.830.0521.176
1.92.0288.530.0951.125
1.81.987.930.0811.185
1.721.899.330.0921.274
1.661.728830.0951.062
1.61.6669.630.1030.998
ReflectionResolution: 1.54→99 Å / Num. all: 44934 / Num. obs: 44934 / % possible obs: 98.3 % / Observed criterion σ(F): 0 / Observed criterion σ(I): 0 / Redundancy: 5.9 % / Biso Wilson estimate: 28.3 Å2 / Rmerge(I) obs: 0.051 / Χ2: 1.59 / Net I/σ(I): 38
Reflection shellResolution: 1.54→1.56 Å / % possible obs: 68.4 % / Rmerge(I) obs: 0.457 / Mean I/σ(I) obs: 3.2 / Num. measured obs: 1526 / Num. unique all: 1526 / Χ2: 1.382 / % possible all: 68.4

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Phasing

PhasingMethod: MAD
Phasing MADD res high: 2 Å / D res low: 19.28 Å / FOM : 0.146 / FOM acentric: 0.142 / FOM centric: 0.17 / Reflection: 10001 / Reflection acentric: 8483 / Reflection centric: 1518
Phasing MAD set

Highest resolution: 2 Å

IDR cullis acentricR cullis centricLowest resolution (Å)Loc acentricLoc centricPower acentricPower centricReflection acentricReflection centric
10.980.96203.650.240.2581751396
20.990.9819.282.13.20.240.2484651516
31.13119.28000084831518
Phasing MAD set shell
IDResolution (Å)R cullis acentricR cullis centricLoc acentricLoc centricPower acentricPower centricReflection acentricReflection centric
19.27-19.281.050.794.25.91.120.785841
16.1-9.270.820.813.34.41.220.8319687
14.55-6.10.910.93.65.50.820.51418134
13.62-4.550.980.995.36.40.390.3636153
13.01-3.620.970.9634.80.40.25975192
12.58-3.010.990.972.53.70.290.181573267
12.25-2.58113.140.130.091888269
12-2.25114.66.50.040.032431253
29.27-19.280.910.9633.91.030.755853
26.1-9.270.860.92.53.61.010.67196100
24.55-6.10.920.942.13.50.860.51418145
23.62-4.550.9812.630.470.38676175
23.01-3.620.990.981.82.70.380.241021210
22.58-3.010.990.981.62.40.240.141573276
22.25-2.58111.72.90.110.061990288
22-2.25112.64.40.030.022533269
39.27-19.282.41100005853
36.1-9.271.8910000196100
34.55-6.11.3710000418145
33.62-4.550.9310000682175
33.01-3.620.93100001024210
32.58-3.010.92100001573276
32.25-2.581.05100001994288
32-2.251100002538271
Phasing MAD set site
IDAtom type symbolB isoFract xFract yFract zOccupancy
1Hg51.458-0.848-0.308-0.1950.029
2Hg63.984-0.31-0.827-0.3330.029
3Hg59.949-0.848-0.308-0.1960.02
4Hg72.973-0.31-0.83-0.3340.018
5Hg81.181-0.849-0.307-0.1970
6Hg86.413-0.31-0.831-0.3340
Phasing MAD shell
Resolution (Å)FOM FOM acentricFOM centricReflectionReflection acentricReflection centric
9.27-19.280.4770.5550.3921115853
6.1-9.270.5260.5910.399296196100
4.55-6.10.4260.4550.344563418145
3.62-4.550.250.2630.2857682175
3.01-3.620.2240.2250.21912341024210
2.58-3.010.1670.1680.16218491573276
2.25-2.580.0660.0670.0622821994288
2-2.250.0230.0240.01928092538271
Phasing dmMethod: Solvent flattening and Histogram matching / Reflection: 10001
Phasing dm shell
Resolution (Å)Delta phi finalFOM Reflection
5.7-10055.90.769504
4.5-5.755.40.764503
3.92-4.558.20.654505
3.45-3.9269.60.301502
3.22-3.4572.30.345501
3.03-3.2270.60.482502
2.88-3.03760.203501
2.76-2.8880.20.455501
2.65-2.76850.33506
2.56-2.6590.20.385507
2.48-2.5689.90.104503
2.41-2.48850.33511
2.35-2.4183.80.18507
2.29-2.3589.10.187512
2.2-2.2994.50.03504
2.15-2.287.80.026512
2.11-2.1583.80.025515
2.07-2.1189.30.021518
2-2.0788.40.249887

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Processing

Software
NameVersionClassificationNB
DENZOdata reduction
SCALEPACKdata scaling
MLPHAREphasing
DM5phasing
REFMACrefmac_5.2.0005refinement
PDB_EXTRACT1.7data extraction
Blu-Icedata collection
RefinementMethod to determine structure: MAD / Resolution: 1.543→19.741 Å / Cor.coef. Fo:Fc: 0.969 / Cor.coef. Fo:Fc free: 0.959 / WRfactor Rfree: 0.207 / WRfactor Rwork: 0.168 / SU B: 2.677 / SU ML: 0.048 / TLS residual ADP flag: LIKELY RESIDUAL / Cross valid method: THROUGHOUT / σ(F): 0 / ESU R: 0.076 / ESU R Free: 0.079 / Stereochemistry target values: Engh & Huber / Details: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS
RfactorNum. reflection% reflectionSelection details
Rfree0.2005 2259 5.033 %random
Rwork0.1652 ---
all0.167 44881 --
obs0.167 44881 99.453 %-
Solvent computationIon probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.2 Å / Solvent model: MASK BULK SOLVENT
Displacement parametersBiso mean: 20.264 Å2
Baniso -1Baniso -2Baniso -3
1-0.008 Å20 Å20 Å2
2---0.001 Å20 Å2
3----0.006 Å2
Refinement stepCycle: LAST / Resolution: 1.543→19.741 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms2461 0 8 229 2698
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.0120.0222500
X-RAY DIFFRACTIONr_bond_other_d0.0010.022416
X-RAY DIFFRACTIONr_angle_refined_deg1.4091.9743332
X-RAY DIFFRACTIONr_angle_other_deg1.44135616
X-RAY DIFFRACTIONr_dihedral_angle_1_deg3.9865302
X-RAY DIFFRACTIONr_dihedral_angle_2_deg21.23923.333120
X-RAY DIFFRACTIONr_dihedral_angle_3_deg12.00815530
X-RAY DIFFRACTIONr_dihedral_angle_4_deg18.8591528
X-RAY DIFFRACTIONr_chiral_restr0.0780.2377
X-RAY DIFFRACTIONr_gen_planes_refined0.0050.022699
X-RAY DIFFRACTIONr_gen_planes_other0.0010.02501
X-RAY DIFFRACTIONr_nbd_refined0.240.2583
X-RAY DIFFRACTIONr_nbd_other0.1760.22467
X-RAY DIFFRACTIONr_nbtor_refined0.1820.21263
X-RAY DIFFRACTIONr_nbtor_other0.0820.21457
X-RAY DIFFRACTIONr_xyhbond_nbd_refined0.1510.2142
X-RAY DIFFRACTIONr_symmetry_vdw_refined0.3170.217
X-RAY DIFFRACTIONr_symmetry_vdw_other0.2680.272
X-RAY DIFFRACTIONr_symmetry_hbond_refined0.1390.226
X-RAY DIFFRACTIONr_symmetry_hbond_other0.0020.21
X-RAY DIFFRACTIONr_mcbond_it2.80221566
X-RAY DIFFRACTIONr_mcbond_other0.7212630
X-RAY DIFFRACTIONr_mcangle_it3.41432439
X-RAY DIFFRACTIONr_mcangle_other2.02232085
X-RAY DIFFRACTIONr_scbond_it3.87621015
X-RAY DIFFRACTIONr_scbond_other1.32321992
X-RAY DIFFRACTIONr_scangle_it6.0813892
X-RAY DIFFRACTIONr_scangle_other2.81433531
LS refinement shell

Refine-ID: X-RAY DIFFRACTION / Total num. of bins used: 20

Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkNum. reflection all% reflection obs (%)
1.543-1.5830.31610.212926328593.973
1.583-1.6260.2541680.18530213189100
1.626-1.6730.2371650.18729403105100
1.673-1.7240.221500.17528623012100
1.724-1.780.2121590.1732766292699.966
1.78-1.8420.2081360.1642702283999.965
1.842-1.9110.2141410.16126042745100
1.911-1.9880.1941380.16124902628100
1.988-2.0750.1711260.15824282554100
2.075-2.1750.1681110.15223362447100
2.175-2.2910.1741060.14822212327100
2.291-2.4280.156870.14421132200100
2.428-2.5930.1861000.15519772077100
2.593-2.7970.191040.15718411945100
2.797-3.0580.227910.16717151806100
3.058-3.4090.2810.1621555163799.939
3.409-3.9170.188980.15313741472100
3.917-4.750.174500.1611202126099.365
4.75-6.5310.227520.215970102599.707
6.531-19.7410.286350.21357964994.607
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)
11.32670.5117-0.70160.8334-0.57922.5876-0.00370.0270.061-0.0359-0.012-0.0164-0.11560.1960.0157-0.0429-0.03710.0071-0.0498-0.0058-0.053520.786719.64631.1151
21.4270.779-0.46131.3415-0.09391.32590.0409-0.0819-0.00020.0524-0.01380.00690.01250.1112-0.0271-0.04560.0088-0.0018-0.0437-0.0059-0.042411.834311.025215.9207
31.49680.7045-0.90971.8692-0.18711.42450.0370.14040.0398-0.1277-0.07370.00910.00880.00560.0367-0.04150.0021-0.0133-0.04330.0238-0.056620.904820.538938.2982
41.94780.9544-0.27211.2284-0.35360.83190.0599-0.06390.00740.0713-0.08590.0394-0.0249-0.01340.026-0.0408-0.02080.0049-0.0517-0.005-0.036211.34811.062752.1324
Refinement TLS group

Refine-ID: X-RAY DIFFRACTION / Selection: ALL

IDRefine TLS-IDAuth asym-IDLabel asym-IDAuth seq-IDLabel seq-ID
11AA1 - 761 - 76
22BB1 - 751 - 75
33CC1 - 761 - 76
44DD1 - 751 - 75

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