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- PDB-2xg8: Structural basis of gene regulation by protein PII: The crystal c... -

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Basic information

Entry
Database: PDB / ID: 2xg8
TitleStructural basis of gene regulation by protein PII: The crystal complex of PII and PipX from Synechococcus elongatus PCC 7942
Components
  • NITROGEN REGULATORY PROTEIN P-II
  • PIPX
KeywordsTRANSCRIPTION / PII SIGNALING PROTEIN / NTCA CO-ACTIVATOR PROTEIN PIPX / TUDOR-LIKE DOMAIN
Function / homology
Function and homology information


regulation of nitrogen utilization / enzyme regulator activity / nucleotide binding / identical protein binding
Similarity search - Function
Protein of unknown function (DUF3539) / Single alpha-helices involved in coiled-coils or other helix-helix interfaces - #870 / PII-interacting protein X, cyanobacteria / PII-interacting protein X / Nitrogen regulatory protein P-II, urydylation site / P-II protein uridylation site. / Nitrogen regulatory protein PII, conserved site / P-II protein C-terminal region signature. / Nitrogen regulatory protein P-II / Nitrogen regulatory protein P-II ...Protein of unknown function (DUF3539) / Single alpha-helices involved in coiled-coils or other helix-helix interfaces - #870 / PII-interacting protein X, cyanobacteria / PII-interacting protein X / Nitrogen regulatory protein P-II, urydylation site / P-II protein uridylation site. / Nitrogen regulatory protein PII, conserved site / P-II protein C-terminal region signature. / Nitrogen regulatory protein P-II / Nitrogen regulatory protein P-II / Nitrogen regulatory protein PII / P-II protein family profile. / Alpha-Beta Plaits - #120 / Nitrogen regulatory PII-like, alpha/beta / Nitrogen regulatory protein PII/ATP phosphoribosyltransferase, C-terminal / Single alpha-helices involved in coiled-coils or other helix-helix interfaces / Helix non-globular / Special / SH3 type barrels. / Roll / Alpha-Beta Plaits / 2-Layer Sandwich / Mainly Beta / Alpha Beta
Similarity search - Domain/homology
Nitrogen regulatory protein P-II / PipX
Similarity search - Component
Biological speciesSYNECHOCOCCUS ELONGATUS (bacteria)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 3.2 Å
AuthorsLlacer, J.L. / Rubio, V.
CitationJournal: Proc.Natl.Acad.Sci.USA / Year: 2010
Title: Structural Basis for the Regulation of Ntca-Dependent Transcription by Proteins Pipx and Pii.
Authors: Llacer, J.L. / Espinosa, J. / Castells, M.A. / Contreras, A. / Forchhammer, K. / Rubio, V.
History
DepositionJun 1, 2010Deposition site: PDBE / Processing site: PDBE
Revision 1.0Aug 18, 2010Provider: repository / Type: Initial release
Revision 1.1May 8, 2011Group: Version format compliance
Revision 1.2Jul 13, 2011Group: Version format compliance
Revision 1.3Dec 20, 2023Group: Data collection / Database references ...Data collection / Database references / Other / Refinement description
Category: chem_comp_atom / chem_comp_bond ...chem_comp_atom / chem_comp_bond / database_2 / pdbx_database_status / pdbx_initial_refinement_model
Item: _database_2.pdbx_DOI / _database_2.pdbx_database_accession / _pdbx_database_status.status_code_sf
Remark 650 HELIX DETERMINATION METHOD: AUTHOR PROVIDED.
Remark 700 SHEET DETERMINATION METHOD: AUTHOR PROVIDED.

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: NITROGEN REGULATORY PROTEIN P-II
B: NITROGEN REGULATORY PROTEIN P-II
C: NITROGEN REGULATORY PROTEIN P-II
D: PIPX
E: PIPX
F: PIPX


Theoretical massNumber of molelcules
Total (without water)68,8516
Polymers68,8516
Non-polymers00
Water543
1
A: NITROGEN REGULATORY PROTEIN P-II
B: NITROGEN REGULATORY PROTEIN P-II
C: NITROGEN REGULATORY PROTEIN P-II
D: PIPX
E: PIPX
F: PIPX

A: NITROGEN REGULATORY PROTEIN P-II
B: NITROGEN REGULATORY PROTEIN P-II
C: NITROGEN REGULATORY PROTEIN P-II
D: PIPX
E: PIPX
F: PIPX


Theoretical massNumber of molelcules
Total (without water)137,70212
Polymers137,70212
Non-polymers00
Water1086
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
crystal symmetry operation2_656-x+1,y,-z+11
Buried area28330 Å2
ΔGint-123.7 kcal/mol
Surface area54520 Å2
MethodPISA
Unit cell
Length a, b, c (Å)111.383, 61.585, 112.979
Angle α, β, γ (deg.)90.00, 126.44, 90.00
Int Tables number5
Space group name H-MC121
Noncrystallographic symmetry (NCS)NCS domain:
IDEns-IDDetails
11A
21B
31C
12A
22C
13D
23E
33F

NCS domain segments:
Dom-IDComponent-IDEns-IDRefine codeAuth asym-IDAuth seq-ID
1111A1 - 36
2111B1 - 36
3111C1 - 36
1211A48 - 100
2211B48 - 100
3211C48 - 100
1314A37 - 43
2314B37 - 43
3314C37 - 43
1413A44 - 47
2413B44 - 47
3413C44 - 47
1513A101
2513B101
3513C101
1611A102 - 105
2611B102 - 105
3611C102 - 105
1713A106 - 108
2713B106 - 108
3713C106 - 108
1124A109 - 112
2124C109 - 112
1131D1 - 7
2131E1 - 7
3131F1 - 7
1232D8
2232E8
3232F8
1331D9 - 19
2331E9 - 19
3331F9 - 19
1432D59
2432E59
3432F59
1531D60 - 62
2531E60 - 62
3531F60 - 62
1631D49 - 58
2631E49 - 58
3631F49 - 58
1734D42 - 47
2734E42 - 47
3734F42 - 47
1832D34
2832E34
3832F34
1931D35 - 41
2931E35 - 41
3931F35 - 41
11034D20 - 25
21034E20 - 25
31034F20 - 25
11131D26 - 33
21131E26 - 33
31131F26 - 33
11232D63
21232E63
31232F63
11331D64 - 65
21331E64 - 65
31331F64 - 65
11432D66 - 67
21432E66 - 67
31432F66 - 67
11534D68 - 70
21534E68 - 70
31534F68 - 70

NCS ensembles :
ID
1
2
3

NCS oper:
IDCodeMatrixVector
1given(-0.4831, 0.6214, -0.6168), (-0.2832, 0.5558, 0.7816), (0.8285, 0.5522, -0.0925)-0.6438, -18.4586, 29.623
2given(-0.461, -0.3084, 0.8321), (0.6209, 0.5579, 0.5507), (-0.634, 0.7705, -0.0657)-29.9665, -5.8087, 16.3787
3given(-0.4795, 0.6309, -0.6099), (-0.2875, 0.5438, 0.7885), (0.8291, 0.5534, -0.0794)-0.4542, -18.5293, 29.2755
4given(-0.4522, -0.2814, 0.8463), (0.66, 0.5326, 0.5298), (-0.5998, 0.7982, -0.0551)-29.5687, -4.3909, 15.998
5given(-0.484, 0.6569, -0.5781), (-0.2541, 0.5267, 0.8112), (0.8374, 0.5395, -0.088)-1.2812, -19.2277, 29.6092
6given(-0.4435, -0.2789, 0.8518), (0.6704, 0.5274, 0.5218), (-0.5948, 0.8025, -0.0469)-30.7316, -4.91, 15.1005

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Components

#1: Protein NITROGEN REGULATORY PROTEIN P-II / PII SIGNAL TRANSDUCING PROTEIN


Mass: 12409.347 Da / Num. of mol.: 3
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) SYNECHOCOCCUS ELONGATUS (bacteria) / Strain: PCC 7942 / Plasmid: PET-22B / Production host: ESCHERICHIA COLI (E. coli) / Strain (production host): BL21(DE3) / References: UniProt: P0A3F4
#2: Protein PIPX


Mass: 10540.987 Da / Num. of mol.: 3
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) SYNECHOCOCCUS ELONGATUS (bacteria) / Strain: PCC 7942 / Plasmid: PTRC99A / Production host: ESCHERICHIA COLI (E. coli) / Strain (production host): BL21(DE3) / References: UniProt: Q7X386
#3: Water ChemComp-HOH / water / Water


Mass: 18.015 Da / Num. of mol.: 3 / 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: 43.82 % / Description: NONE
Crystal growpH: 7.5
Details: PII-PIPX COMPLEX AT 5.5 MG/ML IN 20 MM HEPES PH 7.5, 1MM DTT, 0.4 M NACL, 5 MM MAGNESIUM CHLORIDE. PRECIPITANT SOLUTION: 15 % PEG 3.35K, 0.2 M SODIUM FORMATE.

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: ESRF / Beamline: ID29 / Wavelength: 0.9762
DetectorType: ADSC CCD / Detector: CCD / Date: Nov 10, 2007
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.9762 Å / Relative weight: 1
ReflectionResolution: 3.2→90.9 Å / Num. obs: 10330 / % possible obs: 99.9 % / Observed criterion σ(I): 1.8 / Redundancy: 3.9 % / Biso Wilson estimate: 74.4 Å2 / Rmerge(I) obs: 0.11 / Net I/σ(I): 7.1
Reflection shellResolution: 3→3.37 Å / Redundancy: 3.9 % / Rmerge(I) obs: 0.42 / Mean I/σ(I) obs: 1.8 / % possible all: 99.9

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Processing

Software
NameVersionClassification
REFMAC5.2.0019refinement
MOSFLMdata reduction
SCALAdata scaling
MOLREPphasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: PDB ENTRY 1V5H, CHAIN J
Resolution: 3.2→50 Å / Cor.coef. Fo:Fc: 0.914 / Cor.coef. Fo:Fc free: 0.865 / SU B: 59.859 / SU ML: 0.476 / Cross valid method: THROUGHOUT / ESU R Free: 0.603 / Stereochemistry target values: MAXIMUM LIKELIHOOD
Details: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS. ATOM RECORD CONTAINS SUM OF TLS AND RESIDUAL B FACTORS. ANISOU RECORD CONTAINS SUM OF TLS AND RESIDUAL U FACTORS.
RfactorNum. reflection% reflectionSelection details
Rfree0.27247 495 4.8 %RANDOM
Rwork0.22708 ---
obs0.22923 9829 99.83 %-
Solvent computationIon probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.4 Å / Solvent model: MASK
Displacement parametersBiso mean: 70.104 Å2
Baniso -1Baniso -2Baniso -3
1-1.51 Å20 Å21.6 Å2
2--0.05 Å20 Å2
3---0.35 Å2
Refinement stepCycle: LAST / Resolution: 3.2→50 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms4529 0 0 3 4532
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.0080.0224596
X-RAY DIFFRACTIONr_bond_other_d0.0030.023169
X-RAY DIFFRACTIONr_angle_refined_deg1.2591.9596187
X-RAY DIFFRACTIONr_angle_other_deg1.00937668
X-RAY DIFFRACTIONr_dihedral_angle_1_deg8.255571
X-RAY DIFFRACTIONr_dihedral_angle_2_deg30.79923.514222
X-RAY DIFFRACTIONr_dihedral_angle_3_deg17.69615812
X-RAY DIFFRACTIONr_dihedral_angle_4_deg14.4081543
X-RAY DIFFRACTIONr_chiral_restr0.1420.2707
X-RAY DIFFRACTIONr_gen_planes_refined0.0030.025128
X-RAY DIFFRACTIONr_gen_planes_other0.0020.02995
X-RAY DIFFRACTIONr_nbd_refined0.2280.21037
X-RAY DIFFRACTIONr_nbd_other0.1920.23306
X-RAY DIFFRACTIONr_nbtor_refined0.1810.22213
X-RAY DIFFRACTIONr_nbtor_other0.0830.22711
X-RAY DIFFRACTIONr_xyhbond_nbd_refined0.1220.2124
X-RAY DIFFRACTIONr_xyhbond_nbd_other
X-RAY DIFFRACTIONr_metal_ion_refined
X-RAY DIFFRACTIONr_metal_ion_other
X-RAY DIFFRACTIONr_symmetry_vdw_refined0.2440.239
X-RAY DIFFRACTIONr_symmetry_vdw_other0.2360.277
X-RAY DIFFRACTIONr_symmetry_hbond_refined0.30.23
X-RAY DIFFRACTIONr_symmetry_hbond_other
X-RAY DIFFRACTIONr_symmetry_metal_ion_refined
X-RAY DIFFRACTIONr_symmetry_metal_ion_other
X-RAY DIFFRACTIONr_mcbond_it0.3371.53677
X-RAY DIFFRACTIONr_mcbond_other0.0311.51181
X-RAY DIFFRACTIONr_mcangle_it0.37524567
X-RAY DIFFRACTIONr_mcangle_other
X-RAY DIFFRACTIONr_scbond_it0.47532001
X-RAY DIFFRACTIONr_scbond_other
X-RAY DIFFRACTIONr_scangle_it0.7634.51620
X-RAY DIFFRACTIONr_scangle_other
X-RAY DIFFRACTIONr_long_range_B_refined
X-RAY DIFFRACTIONr_long_range_B_other
X-RAY DIFFRACTIONr_rigid_bond_restr
X-RAY DIFFRACTIONr_sphericity_free
X-RAY DIFFRACTIONr_sphericity_bonded
Refine LS restraints NCS

Refine-ID: X-RAY DIFFRACTION

Ens-IDDom-IDAuth asym-IDNumberTypeRms dev position (Å)Weight position
11A1246tight positional0.020.05
12B1246tight positional0.030.05
13C1246tight positional0.020.05
31D614tight positional0.060.05
32E614tight positional0.030.05
33F614tight positional0.040.05
11A7medium positional1.40.5
12B7medium positional1.380.5
13C7medium positional1.280.5
21A19medium positional0.680.5
31D190medium positional0.950.5
32E190medium positional1.110.5
33F190medium positional0.950.5
11A105loose positional1.365
12B105loose positional1.985
13C105loose positional1.945
11A1246tight thermal0.030.5
12B1246tight thermal0.030.5
13C1246tight thermal0.030.5
31D614tight thermal0.020.5
32E614tight thermal0.030.5
33F614tight thermal0.020.5
11A7medium thermal0.522
12B7medium thermal0.572
13C7medium thermal0.182
21A19medium thermal0.132
31D190medium thermal0.132
32E190medium thermal0.142
33F190medium thermal0.122
11A105loose thermal0.4310
12B105loose thermal0.3310
13C105loose thermal0.4610
LS refinement shellResolution: 3.2→3.283 Å / Total num. of bins used: 20
RfactorNum. reflection% reflection
Rfree0.323 30 -
Rwork0.259 710 -
obs--100 %
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)
14.65261.1886-0.6225.6623-1.23495.18090.07640.22940.11060.1117-0.0477-0.0232-0.1424-0.2051-0.0286-0.380.04680.0506-0.3439-0.0382-0.4048-12.8215.35112.371
25.2084-0.9108-2.36765.26040.49095.14330.0902-0.23510.04070.0515-0.14730.16050.2152-0.02220.0571-0.4378-0.1268-0.0101-0.2650.0498-0.3796-15.659-3.16927.716
33.770.03130.98672.42930.89585.7731-0.1134-0.4964-0.03360.1060.0078-0.15030.47190.28430.1057-0.4342-0.02160.0714-0.25090.0415-0.25721.365-2.30621.043
49.9472-1.3156-3.79665.01313.35027.01720.3679-0.92241.54610.04230.1742-0.4165-0.43170.1416-0.5422-0.3138-0.14050.0456-0.0207-0.1643-0.0611-14.21213.23644.317
55.4406-0.42820.11199.1768-0.58587.07030.24650.10490.77450.363-0.218-0.2602-0.5540.3409-0.02840.0188-0.01590.1649-0.3669-0.0309-0.0651-12.12527.84520.703
64.3751.20531.05775.1446-0.92922.1687-0.3056-0.18281.08190.5228-0.10550.3143-0.33530.59570.4111-0.1561-0.25170.04160.0992-0.13470.208910.41616.41332.297
713.88556.72191.314210.05050.40375.54370.5611-0.0037-0.35880.3062-0.38930.32750.90140.0995-0.1719-0.270.10820.1072-0.0742-0.0633-0.1253-43.0866.21845.995
831.559417.3912-5.807311.7388-3.061512.3165-1.89291.435-0.8826-1.63550.71620.55711.11850.20471.17680.4813-0.0281-0.15520.02440.0320.1701-14.33231.7096.968
920.2741-9.035518.09624.0268-8.064816.15221.8708-0.01272.409-0.1987-2.7889-2.36893.0054-0.86550.91820.1368-0.06090.17640.18010.12930.542825.1699.90864.204
Refinement TLS group
IDRefine-IDRefine TLS-IDAuth asym-IDAuth seq-ID
1X-RAY DIFFRACTION1A1 - 110
2X-RAY DIFFRACTION2B1 - 108
3X-RAY DIFFRACTION3C1 - 112
4X-RAY DIFFRACTION4D3 - 70
5X-RAY DIFFRACTION5E3 - 70
6X-RAY DIFFRACTION6F3 - 70
7X-RAY DIFFRACTION7D71 - 89
8X-RAY DIFFRACTION8E71 - 89
9X-RAY DIFFRACTION9F79 - 89

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