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- PDB-3nke: High resolution structure of the C-terminal domain CRISP-associat... -

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Basic information

Entry
Database: PDB / ID: 3nke
TitleHigh resolution structure of the C-terminal domain CRISP-associated protein Cas1 from Escherichia coli str. K-12
Componentsprotein ygbT
KeywordsIMMUNE SYSTEM / CRISPR / Cas1 / YgbT / Nuclease / DNA recombination / DNA repair
Function / homology
Function and homology information


CRISPR-cas system / crossover junction DNA endonuclease activity / 5'-flap endonuclease activity / maintenance of CRISPR repeat elements / defense response to virus / Hydrolases; Acting on ester bonds / DNA repair / DNA damage response / protein homodimerization activity / DNA binding ...CRISPR-cas system / crossover junction DNA endonuclease activity / 5'-flap endonuclease activity / maintenance of CRISPR repeat elements / defense response to virus / Hydrolases; Acting on ester bonds / DNA repair / DNA damage response / protein homodimerization activity / DNA binding / identical protein binding / metal ion binding / cytoplasm
Similarity search - Function
CRISPR-associated protein Cas1, ECOLI subtype / CRISPR-associated protein Cas1, type I-E / CRISPR-associated endonuclease Cas1, C-terminal domain / CRISPR-associated endonuclease Cas1, N-terminal domain / : / CRISPR-associated protein Cas1 / CRISPR-associated endonuclease Cas1, C-terminal domain / CRISPR associated protein Cas1 / Four Helix Bundle (Hemerythrin (Met), subunit A) / Up-down Bundle / Mainly Alpha
Similarity search - Domain/homology
SULFITE ION / CRISPR-associated endonuclease Cas1
Similarity search - Component
Biological speciesEscherichia coli (E. coli)
MethodX-RAY DIFFRACTION / SYNCHROTRON / Resolution: 1.4 Å
AuthorsNocek, B. / Skarina, T. / Beloglazova, N. / Savchenko, A. / Joachimiak, A. / Yakunin, A.
CitationJournal: Mol.Microbiol. / Year: 2011
Title: A dual function of the CRISPR-Cas system in bacterial antivirus immunity and DNA repair.
Authors: Babu, M. / Beloglazova, N. / Flick, R. / Graham, C. / Skarina, T. / Nocek, B. / Gagarinova, A. / Pogoutse, O. / Brown, G. / Binkowski, A. / Phanse, S. / Joachimiak, A. / Koonin, E.V. / ...Authors: Babu, M. / Beloglazova, N. / Flick, R. / Graham, C. / Skarina, T. / Nocek, B. / Gagarinova, A. / Pogoutse, O. / Brown, G. / Binkowski, A. / Phanse, S. / Joachimiak, A. / Koonin, E.V. / Savchenko, A. / Emili, A. / Greenblatt, J. / Edwards, A.M. / Yakunin, A.F.
History
DepositionJun 18, 2010Deposition site: RCSB / Processing site: RCSB
Revision 1.0Aug 25, 2010Provider: repository / Type: Initial release
Revision 1.1Jul 13, 2011Group: Version format compliance
Revision 1.2Dec 17, 2014Group: Data collection
Revision 1.3Oct 30, 2024Group: Data collection / Database references ...Data collection / Database references / Derived calculations / Structure summary
Category: chem_comp_atom / chem_comp_bond ...chem_comp_atom / chem_comp_bond / database_2 / pdbx_entry_details / pdbx_modification_feature / struct_conn / struct_site
Item: _database_2.pdbx_DOI / _database_2.pdbx_database_accession ..._database_2.pdbx_DOI / _database_2.pdbx_database_accession / _struct_conn.pdbx_leaving_atom_flag / _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: protein ygbT
B: protein ygbT
C: protein ygbT
hetero molecules


Theoretical massNumber of molelcules
Total (without water)67,24414
Polymers66,3553
Non-polymers88911
Water13,836768
1


  • Idetical with deposited unit
  • defined by author&software
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area7100 Å2
ΔGint-84 kcal/mol
Surface area23920 Å2
MethodPISA
Unit cell
Length a, b, c (Å)53.709, 90.047, 121.876
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 protein ygbT / CRISP-associated protein Cas1


Mass: 22118.281 Da / Num. of mol.: 3 / Fragment: UNP residues 92-291
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Escherichia coli (E. coli) / Strain: K-12 / Gene: ygbT, b2755, JW2725 / Production host: Escherichia coli (E. coli) / Strain (production host): BL21DE3 / References: UniProt: Q46896
#2: Chemical
ChemComp-EDO / 1,2-ETHANEDIOL / ETHYLENE GLYCOL


Mass: 62.068 Da / Num. of mol.: 4 / Source method: obtained synthetically / Formula: C2H6O2
#3: Chemical ChemComp-SO3 / SULFITE ION


Mass: 80.063 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: SO3
#4: Chemical
ChemComp-SO4 / SULFATE ION


Mass: 96.063 Da / Num. of mol.: 5 / Source method: obtained synthetically / Formula: SO4
#5: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 768 / Source method: isolated from a natural source / Formula: H2O
Has protein modificationY

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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: 2.23 Å3/Da / Density % sol: 44.73 %
Crystal growTemperature: 291 K / Method: vapor diffusion, hanging drop / pH: 5.6
Details: O.O5 M MES 20% PEG 8k, 0.01 M Magnesium Chloride, 0.01M Adenelyl-imidodophosphate, 0.2M Ammonium sulfate, 0.3M NDSB211, pH 5.6, VAPOR DIFFUSION, HANGING DROP, temperature 291K

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: APS / Beamline: 19-ID / Wavelength: 0.9794 Å
DetectorType: ADSC QUANTUM 315r / Detector: CCD / Date: Apr 20, 2010 / Details: mirrors
RadiationMonochromator: double crystal / Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.9794 Å / Relative weight: 1
ReflectionResolution: 1.4→45 Å / Num. all: 117000 / Num. obs: 115817 / % possible obs: 99 % / Observed criterion σ(F): 2 / Observed criterion σ(I): 2 / Redundancy: 4.8 % / Biso Wilson estimate: 22 Å2 / Rmerge(I) obs: 0.065 / Net I/σ(I): 28
Reflection shellResolution: 1.4→1.42 Å / Redundancy: 4.7 % / Rmerge(I) obs: 0.64 / Mean I/σ(I) obs: 2.5 / Num. unique all: 5726

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Processing

Software
NameVersionClassification
SBC-Collectdata collection
HKL-3000phasing
REFMAC5.5.0109refinement
HKL-3000data reduction
HKL-3000data scaling
RefinementResolution: 1.4→33.2 Å / Cor.coef. Fo:Fc: 0.962 / Cor.coef. Fo:Fc free: 0.952 / SU B: 1.997 / SU ML: 0.037 / Cross valid method: THROUGHOUT / ESU R: 0.061 / ESU R Free: 0.062 / Stereochemistry target values: MAXIMUM LIKELIHOOD / Details: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS
RfactorNum. reflection% reflectionSelection details
Rfree0.20067 5778 5 %RANDOM
Rwork0.17656 ---
obs0.17777 109670 98.77 %-
Solvent computationIon probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.4 Å / Solvent model: MASK
Displacement parametersBiso mean: 14.603 Å2
Baniso -1Baniso -2Baniso -3
1-0 Å20 Å20 Å2
2--0.55 Å20 Å2
3----0.56 Å2
Refinement stepCycle: LAST / Resolution: 1.4→33.2 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms4235 0 49 768 5052
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.0150.0224457
X-RAY DIFFRACTIONr_bond_other_d0.0010.023172
X-RAY DIFFRACTIONr_angle_refined_deg1.5911.9936043
X-RAY DIFFRACTIONr_angle_other_deg1.1337681
X-RAY DIFFRACTIONr_dihedral_angle_1_deg4.4015576
X-RAY DIFFRACTIONr_dihedral_angle_2_deg32.00821.872187
X-RAY DIFFRACTIONr_dihedral_angle_3_deg11.25315771
X-RAY DIFFRACTIONr_dihedral_angle_4_deg15.1731553
X-RAY DIFFRACTIONr_chiral_restr0.0920.2685
X-RAY DIFFRACTIONr_gen_planes_refined0.0080.0214935
X-RAY DIFFRACTIONr_gen_planes_other0.0020.02947
X-RAY DIFFRACTIONr_nbd_refined
X-RAY DIFFRACTIONr_nbd_other
X-RAY DIFFRACTIONr_nbtor_refined
X-RAY DIFFRACTIONr_nbtor_other
X-RAY DIFFRACTIONr_xyhbond_nbd_refined
X-RAY DIFFRACTIONr_xyhbond_nbd_other
X-RAY DIFFRACTIONr_metal_ion_refined
X-RAY DIFFRACTIONr_metal_ion_other
X-RAY DIFFRACTIONr_symmetry_vdw_refined
X-RAY DIFFRACTIONr_symmetry_vdw_other
X-RAY DIFFRACTIONr_symmetry_hbond_refined
X-RAY DIFFRACTIONr_symmetry_hbond_other
X-RAY DIFFRACTIONr_symmetry_metal_ion_refined
X-RAY DIFFRACTIONr_symmetry_metal_ion_other
X-RAY DIFFRACTIONr_mcbond_it0.91.52791
X-RAY DIFFRACTIONr_mcbond_other0.2321.51124
X-RAY DIFFRACTIONr_mcangle_it1.60824492
X-RAY DIFFRACTIONr_scbond_it2.55931666
X-RAY DIFFRACTIONr_scangle_it4.1174.51537
X-RAY DIFFRACTIONr_rigid_bond_restr
X-RAY DIFFRACTIONr_sphericity_free
X-RAY DIFFRACTIONr_sphericity_bonded
LS refinement shellResolution: 1.4→1.436 Å / Total num. of bins used: 20
RfactorNum. reflection% reflection
Rfree0.262 419 -
Rwork0.248 8048 -
obs--99.06 %
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)
10.83420.41160.00950.52360.76521.234-0.0308-0.00320.02330.0363-0.05110.0294-0.0076-0.18030.0820.09410.0242-0.00430.1115-0.01890.09873.96535.510631.4182
22.00960.0269-1.01280.22660.18510.6089-0.05690.1112-0.05610.0343-0.01040.01950.0529-0.04690.06730.0966-0.01570.00770.1045-0.01940.09689.7977-3.231518.1898
30.87960.5716-0.17411.5722-0.67570.4548-0.0140.0886-0.0637-0.13450.00460.01910.07590.00190.00940.1151-0.01390.00350.0978-0.03060.077212.1144-8.592711.9092
42.88790.0227-0.8190.61910.3790.28490.0836-0.1036-0.14940.0731-0.0857-0.03890.03910.00330.00210.1142-0.0103-0.00410.075-0.00440.11217.2676-11.828319.2014
51.43090.5120.32890.3419-0.18610.5120.04130.101-0.0971-0.03780.03190.00810.10180.0709-0.07320.09320.0168-0.0060.0951-0.03240.102434.0032-8.063516.3383
63.0009-6.031-5.575719.41824.31714.73750.018-0.4849-0.53110.8675-0.2728-0.11870.07350.08880.25480.1641-0.1113-0.1130.25150.10250.097434.7008-9.404730.8828
71.4595-1.6302-0.0872.16710.31320.2592-0.0205-0.1120.02070.11350.042-0.05580.028-0.0034-0.02140.07230.0151-0.0150.1224-0.02370.103336.0623-1.854227.8846
80.96630.3275-0.450.70610.40310.95-0.0403-0.07870.06940.03250.03370.02460.05870.06560.00660.07580.0171-0.01040.0958-0.0220.095616.93284.387530.332
90.80910.0467-0.4025-0.00070.11170.451-0.0481-0.02410.0190.01790.0512-0.00970.06070.0388-0.00310.08960.0126-0.00520.1071-0.01560.10224.92210.657224.7158
100.1694-0.7598-0.45651.80320.84031.6915-0.07720.07170.05920.06680.0434-0.06870.02580.07650.03390.041-0.0074-0.01450.1595-0.02660.117145.0392.880424.0471
112.2482-1.09080.55650.68490.14010.6283-0.01040.0727-0.18440.01140.11190.0954-0.10840.1396-0.10150.08480.00170.00230.1037-0.01230.114135.712711.781726.7084
122.9261.1563-0.07590.38080.05020.3839-0.04010.16140.1723-0.00450.07640.0552-0.0509-0.0368-0.03630.07730.0078-0.01480.09630.00720.123812.332910.376121.4116
132.234-0.2675-1.05090.1876-0.60982.4739-0.0380.01870.02750.0652-0.0082-0.0036-0.3007-0.12540.04610.14220.0575-0.00250.07940.01010.08739.038316.50947.6926
140.53310.23910.16460.8832-0.17790.23640.07360.01250.01270.0085-0.08870.0115-0.0262-0.09320.01520.09250.02870.00870.1419-0.01030.07225.09852.064159.44
152.84612.73844.28241.79793.27424.90250.08980.1179-0.20070.10270.11460.00130.03180.1775-0.20440.07580.00010.02420.1351-0.01830.14320.0347-6.226567.2719
16-0.15050.55260.06811.53231.14092.16350.0037-0.00610.00640.1150.0182-0.02380.21680.1131-0.02190.13190.0271-0.01370.0881-0.03170.08297.6779-12.49359.8769
174.15481.0129-0.27471.246-0.46463.4402-0.063-0.1018-0.01140.1073-0.0010.04230.5119-0.05280.0640.1948-0.01150.02190.0041-0.02310.05046.4083-24.695347.5119
180.1764-1.19450.06873.11280.18871.1503-0.0371-0.01160.03550.06480.0519-0.04830.0927-0.0119-0.01470.13030.0164-0.00050.0566-0.01860.101911.7789-15.078340.5863
190.38880.54080.09641.15290.04480.59450.05510.0386-0.04270.0281-0.0286-0.0875-0.0145-0.0201-0.02640.08850.03130.00450.1057-0.0030.087110.62781.069247.9057
200.7972-0.8028-0.08512.75480.5440.0609-0.0217-0.0627-0.02850.03170.01260.10970.153-0.20220.0090.1517-0.08610.01040.141-0.01190.092.15-18.626941.2277
214.1469-3.3601-5.17815.68413.04373.0104-0.12740.0739-0.0748-0.0240.04930.07680.1754-0.03270.07820.13530.03510.01020.0686-0.02910.08789.9472-20.552331.0473
223.2324-3.45120.84054.0244-1.33211.2162-0.07460.01510.09150.08910.0313-0.06160.1212-0.11430.04330.07980.003-0.00540.0842-0.00980.10441.5045-9.528333.4937
230.44570.18650.89361.78762.25043.56020.0215-0.02380.0112-0.0743-0.09690.0535-0.0375-0.09980.07540.07840.06110.00390.1493-0.01240.0673-1.53983.254845.8324
2412.66624.4206-1.89118.9493-5.036412.03070.3353-0.04650.7320.0649-0.80710.2508-0.4953-0.66440.47170.07290.21250.01810.2182-0.05140.07050.983213.959856.1769
251.12020.53330.85961.406-0.06110.1713-0.0124-0.04150.004-0.04630.01740.1619-0.08640.0164-0.0050.12370.0231-0.02070.07230.00110.121220.436419.704232.1361
260.24450.10220.3060.3488-0.34191.8131-0.01030.02040.00890.0356-0.0228-0.0266-0.05920.04190.03310.1128-0.0127-0.010.0857-0.00040.091636.391520.43642.0613
275.1023-4.8163-6.89684.46887.376916.70110.2704-0.1840.75460.08480.319-0.8238-0.10660.4657-0.58940.1398-0.0367-0.05450.0492-0.03480.244446.301822.285843.5913
282.2704-0.61320.36240.1273-0.03624.1884-0.05690.05930.0580.0043-0.0135-0.00540.24260.15170.07040.10220.0204-0.00060.09980.0140.078143.324212.811740.4964
291.00090.6996-0.750.3213-0.12010.77330.0254-0.1074-0.0429-0.0258-0.0516-0.01360.00230.1820.02620.08510.05410.00520.1570.03160.056442.04255.662358.1761
306.66821.0809-2.18663.9036-0.45011.46850.05810.3649-0.71080.0547-0.0147-0.08240.130.1108-0.04340.080.0523-0.02650.1109-0.02660.170935.5044-4.189156.3076
31-0.0557-0.11510.21750.5909-0.58731.65520.0249-0.0139-0.0616-0.00220.06190.0953-0.0152-0.0171-0.08680.08260.0109-0.00840.08960.02860.115824.17988.994646.0215
320.0385-0.05270.67240.65270.08342.31510.06840.0104-0.0703-0.00520.08140.05890.0790.1414-0.14980.10770.0009-0.02220.0848-0.00830.110329.19629.356743.456
330.3729-0.6271.28711.3521-1.42662.34640.0226-0.0479-0.00030.03340.01840.0487-0.02790.0351-0.0410.09550.0053-0.00190.12010.04120.088932.03485.700363.3934
340.8049-0.54880.61082.7591-0.78580.37840.0991-0.0745-0.09840.0397-0.06360.0044-0.0321-0.0712-0.03550.11340.01980.01670.09020.04110.108921.2625.148662.5221
350.93560.4495-2.11281.4193-1.17783.3813-0.01410.0126-0.03540.1290.04430.0636-0.11910.0461-0.03020.14560.01930.01640.05680.01070.103123.589121.747547.1618
3617.1251-1.17274.62226.29951.8021.19960.1610.09770.19450.1205-0.34230.4783-0.33550.16340.18130.3302-0.01190.01570.0031-0.01520.090523.942728.137735.0841
Refinement TLS group
IDRefine-IDRefine TLS-IDAuth asym-IDAuth seq-ID
1X-RAY DIFFRACTION1A94 - 108
2X-RAY DIFFRACTION2A109 - 123
3X-RAY DIFFRACTION3A124 - 134
4X-RAY DIFFRACTION4A135 - 143
5X-RAY DIFFRACTION5A144 - 163
6X-RAY DIFFRACTION6A164 - 172
7X-RAY DIFFRACTION7A173 - 186
8X-RAY DIFFRACTION8A187 - 208
9X-RAY DIFFRACTION9A209 - 234
10X-RAY DIFFRACTION10A235 - 244
11X-RAY DIFFRACTION11A245 - 257
12X-RAY DIFFRACTION12A258 - 280
13X-RAY DIFFRACTION13B92 - 101
14X-RAY DIFFRACTION14B102 - 124
15X-RAY DIFFRACTION15B125 - 131
16X-RAY DIFFRACTION16B132 - 151
17X-RAY DIFFRACTION17B152 - 164
18X-RAY DIFFRACTION18B172 - 190
19X-RAY DIFFRACTION19B191 - 223
20X-RAY DIFFRACTION20B224 - 238
21X-RAY DIFFRACTION21B239 - 246
22X-RAY DIFFRACTION22B247 - 258
23X-RAY DIFFRACTION23B259 - 274
24X-RAY DIFFRACTION24B275 - 281
25X-RAY DIFFRACTION25C92 - 107
26X-RAY DIFFRACTION26C108 - 125
27X-RAY DIFFRACTION27C126 - 132
28X-RAY DIFFRACTION28C133 - 140
29X-RAY DIFFRACTION29C141 - 162
30X-RAY DIFFRACTION30C163 - 178
31X-RAY DIFFRACTION31C179 - 207
32X-RAY DIFFRACTION32C208 - 223
33X-RAY DIFFRACTION33C224 - 238
34X-RAY DIFFRACTION34C239 - 261
35X-RAY DIFFRACTION35C262 - 274
36X-RAY DIFFRACTION36C275 - 281

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