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- PDB-2pmu: Crystal structure of the DNA-binding domain of PhoP -

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Basic information

Entry
Database: PDB / ID: 2pmu
TitleCrystal structure of the DNA-binding domain of PhoP
Componentsresponse regulator PHOP
KeywordsTRANSCRIPTION REGULATION / winged helix-tun-helx
Function / homology
Function and homology information


protein secretion by the type VII secretion system / glycolipid biosynthetic process / phosphorelay response regulator activity / phosphorelay signal transduction system / positive regulation of lipid biosynthetic process / protein-DNA complex / response to oxidative stress / transcription cis-regulatory region binding / negative regulation of DNA-templated transcription / positive regulation of gene expression ...protein secretion by the type VII secretion system / glycolipid biosynthetic process / phosphorelay response regulator activity / phosphorelay signal transduction system / positive regulation of lipid biosynthetic process / protein-DNA complex / response to oxidative stress / transcription cis-regulatory region binding / negative regulation of DNA-templated transcription / positive regulation of gene expression / regulation of DNA-templated transcription / positive regulation of DNA-templated transcription / DNA binding / metal ion binding / plasma membrane / cytosol
Similarity search - Function
OmpR/PhoB-type DNA-binding domain profile. / OmpR/PhoB-type DNA-binding domain / Transcriptional regulatory protein, C terminal / Transcriptional regulatory protein, C terminal / Transcriptional regulatory protein WalR-like / Signal transduction response regulator, C-terminal effector / Response regulator receiver domain / cheY-homologous receiver domain / Signal transduction response regulator, receiver domain / Response regulatory domain profile. ...OmpR/PhoB-type DNA-binding domain profile. / OmpR/PhoB-type DNA-binding domain / Transcriptional regulatory protein, C terminal / Transcriptional regulatory protein, C terminal / Transcriptional regulatory protein WalR-like / Signal transduction response regulator, C-terminal effector / Response regulator receiver domain / cheY-homologous receiver domain / Signal transduction response regulator, receiver domain / Response regulatory domain profile. / CheY-like superfamily / Winged helix-like DNA-binding domain superfamily/Winged helix DNA-binding domain / Arc Repressor Mutant, subunit A / Winged helix-like DNA-binding domain superfamily / Orthogonal Bundle / Mainly Alpha
Similarity search - Domain/homology
GLYCINE / : / PHOSPHATE ION / Possible two component system response transcriptional positive regulator PhoP / DNA-binding response regulator
Similarity search - Component
Biological speciesMycobacterium tuberculosis (bacteria)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 1.779 Å
AuthorsWang, S.
CitationJournal: Biochemistry / Year: 2007
Title: Structure of the DNA-binding domain of the response regulator PhoP from Mycobacterium tuberculosis
Authors: Wang, S. / Engohang-Ndong, J. / Smith, I.
History
DepositionApr 23, 2007Deposition site: RCSB / Processing site: RCSB
Revision 1.0Feb 26, 2008Provider: repository / Type: Initial release
Revision 1.1Jul 13, 2011Group: Advisory / Source and taxonomy / Version format compliance
Revision 1.2Oct 18, 2017Group: Refinement description / Category: software
Revision 1.3Aug 30, 2023Group: Data collection / Database references ...Data collection / Database references / Derived calculations / Refinement description
Category: chem_comp_atom / chem_comp_bond ...chem_comp_atom / chem_comp_bond / database_2 / pdbx_initial_refinement_model / struct_ncs_dom_lim / struct_ref_seq_dif / struct_site
Item: _database_2.pdbx_DOI / _database_2.pdbx_database_accession ..._database_2.pdbx_DOI / _database_2.pdbx_database_accession / _struct_ncs_dom_lim.beg_auth_comp_id / _struct_ncs_dom_lim.end_auth_comp_id / _struct_ref_seq_dif.details / _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: response regulator PHOP
B: response regulator PHOP
C: response regulator PHOP
D: response regulator PHOP
E: response regulator PHOP
F: response regulator PHOP
hetero molecules


Theoretical massNumber of molelcules
Total (without water)77,60824
Polymers76,7446
Non-polymers86418
Water8,809489
1


  • Idetical with deposited unit
  • defined by author
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Unit cell
Length a, b, c (Å)103.884, 101.129, 86.930
Angle α, β, γ (deg.)90.00, 126.72, 90.00
Int Tables number5
Space group name H-MC121
Noncrystallographic symmetry (NCS)NCS domain:
IDEns-IDDetails
11A
21D
31A
41D
51A
61D
71A
81D
91A
101D
111A
121D
131A
141D
151A
161D
171A
181D
191A
201D
211A
221D
12B
22E
32B
42E
52B
62E
72B
82E
92B
102E
112B
122E
132B
142E
152B
162E
172B
182E
192B
202E
212B
222E
232B
242E
252B
262E
272B
282E
292B
302E
312B
322E
332B
342E
13C
23F
33C
43F
53C
63F
73C
83F
93C
103F

NCS domain segments:
Dom-IDComponent-IDEns-IDBeg auth comp-IDBeg label comp-IDEnd auth comp-IDEnd label comp-IDRefine codeAuth asym-IDLabel asym-IDAuth seq-IDLabel seq-ID
111LEULEUGLUGLU1AA151 - 17812 - 39
211LEULEUGLUGLU1DD151 - 17812 - 39
321VALVALLYSLYS1AA213 - 23074 - 91
421VALVALLYSLYS1DD213 - 23074 - 91
531VALVALARGARG1AA239 - 244100 - 105
631VALVALARGARG1DD239 - 244100 - 105
741PHEPHEPHEPHE3AA17940
841PHEPHEPHEPHE3DD17940
951ARGARGARGARG3AA20465
1051ARGARGARGARG3DD20465
1161ARGARGARGARG3AA23192
1261ARGARGARGARG3DD23192
1371LEULEULEULEU3AA23394
1471LEULEULEULEU3DD23394
1581THRTHRTRPTRP1AA180 - 20341 - 64
1681THRTHRTRPTRP1DD180 - 20341 - 64
1791TYRTYRTYRTYR1AA20566
1891TYRTYRTYRTYR1DD20566
19101LEULEULEULEU1AA23293
20101LEULEULEULEU1DD23293
21111HISHISLEULEU1AA234 - 23695 - 97
22111HISHISLEULEU1DD234 - 23695 - 97
112PROPROVALVAL1BB146 - 1497 - 10
212PROPROVALVAL1EE146 - 1497 - 10
322HISHISASPASP1BB201 - 20662 - 67
422HISHISASPASP1EE201 - 20662 - 67
532ASNASNASNASN1BB21273
632ASNASNASNASN1EE21273
742LEULEULEULEU1BB232 - 23693 - 97
842LEULEULEULEU1EE232 - 23693 - 97
952GLYGLYGLYGLY1BB240101
1052GLYGLYGLYGLY1EE240101
1162ARGARGARGARG3BB15011
1262ARGARGARGARG3EE15011
1372GLUGLUGLUGLU3BB16021
1472GLUGLUGLUGLU3EE16021
1582PHEPHEPHEPHE3BB17940
1682PHEPHEPHEPHE3EE17940
1792VALVALVALVAL3BB21374
1892VALVALVALVAL3EE21374
19102TYRTYRTYRTYR3BB241102
20102TYRTYRTYRTYR3EE241102
21112LEULEULEULEU3BB243104
22112LEULEULEULEU3EE243104
23122LEULEUASPASP1BB151 - 15912 - 20
24122LEULEUASPASP1EE151 - 15912 - 20
25132GLUGLUGLUGLU1BB161 - 17822 - 39
26132GLUGLUGLUGLU1EE161 - 17822 - 39
27142THRTHRLEULEU1BB180 - 19941 - 60
28142THRTHRLEULEU1EE180 - 19941 - 60
29152VALVALASPASP1BB214 - 22675 - 87
30152VALVALASPASP1EE214 - 22675 - 87
31162VALVALVALVAL1BB242103
32162VALVALVALVAL1EE242103
33172ARGARGARGARG1BB244105
34172ARGARGARGARG1EE244105
113LEULEUASPASP1CC151 - 15912 - 20
213LEULEUASPASP1FF151 - 15912 - 20
323THRTHRTRPTRP1CC162 - 20323 - 64
423THRTHRTRPTRP1FF162 - 20323 - 64
533VALVALGLYGLY1CC213 - 22874 - 89
633VALVALGLYGLY1FF213 - 22874 - 89
743GLYGLYLEULEU1CC240 - 243101 - 104
843GLYGLYLEULEU1FF240 - 243101 - 104
953ARGARGHISHIS1CC231 - 23492 - 95
1053ARGARGHISHIS1FF231 - 23492 - 95

NCS ensembles :
ID
1
2
3

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Components

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Protein , 1 types, 6 molecules ABCDEF

#1: Protein
response regulator PHOP / POSSIBLE TWO COMPONENT SYSTEM RESPONSE TRANSCRIPTIONAL POSITIVE REGULATOR PHOP


Mass: 12790.662 Da / Num. of mol.: 6 / Fragment: DNA-binding domain, residues 144-247
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Mycobacterium tuberculosis (bacteria) / Strain: H37Rv / Gene: PhoP / Plasmid: pET28 / Species (production host): Escherichia coli / Production host: Escherichia coli BL21(DE3) (bacteria) / Strain (production host): BL21(DE3) / References: UniProt: P71814, UniProt: Q7D9B8*PLUS

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Non-polymers , 6 types, 507 molecules

#2: Chemical
ChemComp-PO4 / PHOSPHATE ION


Mass: 94.971 Da / Num. of mol.: 4 / Source method: obtained synthetically / Formula: PO4
#3: Chemical ChemComp-K / POTASSIUM ION


Mass: 39.098 Da / Num. of mol.: 3 / Source method: obtained synthetically / Formula: K
#4: Chemical
ChemComp-UNX / UNKNOWN ATOM OR ION


Num. of mol.: 4 / Source method: obtained synthetically
#5: Chemical
ChemComp-CL / CHLORIDE ION


Mass: 35.453 Da / Num. of mol.: 4 / Source method: obtained synthetically / Formula: Cl
#6: Chemical ChemComp-GLY / GLYCINE


Type: peptide linking / Mass: 75.067 Da / Num. of mol.: 3 / Source method: obtained synthetically / Formula: C2H5NO2
#7: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 489 / 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: 2.38 Å3/Da / Density % sol: 48.4 %
Crystal growTemperature: 298 K / Method: vapor diffusion / pH: 5.6
Details: 1.4 M Na/K phosphate, pH 5.6, 100 mM glycine, VAPOR DIFFUSION, temperature 298K

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: NSLS / Beamline: X6A / Wavelength: 0.9795 Å
DetectorType: ADSC QUANTUM 210 / Detector: CCD / Date: Oct 8, 2006
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.9795 Å / Relative weight: 1
ReflectionResolution: 1.779→20 Å / Num. obs: 65220 / % possible obs: 94.8 % / Redundancy: 2.9 % / Rmerge(I) obs: 0.036 / Χ2: 1.033 / Net I/σ(I): 18.6
Reflection shellResolution: 1.779→1.84 Å / Redundancy: 2.7 % / Rmerge(I) obs: 0.582 / Mean I/σ(I) obs: 2.03 / Num. unique all: 6629 / Χ2: 1.05 / % possible all: 96.7

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Processing

Software
NameVersionClassificationNB
DENZOdata reduction
SCALEPACKdata scaling
REFMAC5.2.0019refinement
PDB_EXTRACT2data extraction
PHASERphasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: PDB entry 1KGS C-terminal domain
Resolution: 1.779→20 Å / Cor.coef. Fo:Fc: 0.956 / Cor.coef. Fo:Fc free: 0.935 / SU B: 5.454 / SU ML: 0.09 / TLS residual ADP flag: LIKELY RESIDUAL / Isotropic thermal model: isotopic / Cross valid method: THROUGHOUT / σ(F): 0 / ESU R: 0.131 / ESU R Free: 0.129 / Stereochemistry target values: MAXIMUM LIKELIHOOD / Details: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS
RfactorNum. reflection% reflectionSelection details
Rfree0.238 3310 5.1 %RANDOM
Rwork0.196 ---
obs0.198 65210 94.63 %-
Solvent computationIon probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.4 Å / Solvent model: BABINET MODEL WITH MASK
Displacement parametersBiso mean: 41.509 Å2
Baniso -1Baniso -2Baniso -3
1-0.35 Å20 Å20.43 Å2
2---0.48 Å20 Å2
3---0.64 Å2
Refinement stepCycle: LAST / Resolution: 1.779→20 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms4796 0 46 489 5331
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.0160.0224966
X-RAY DIFFRACTIONr_angle_refined_deg1.5541.976748
X-RAY DIFFRACTIONr_dihedral_angle_1_deg5.7195588
X-RAY DIFFRACTIONr_dihedral_angle_2_deg34.00622.361233
X-RAY DIFFRACTIONr_dihedral_angle_3_deg15.7315856
X-RAY DIFFRACTIONr_dihedral_angle_4_deg19.7341549
X-RAY DIFFRACTIONr_chiral_restr0.1170.2764
X-RAY DIFFRACTIONr_gen_planes_refined0.0080.023711
X-RAY DIFFRACTIONr_nbd_refined0.2360.22286
X-RAY DIFFRACTIONr_nbtor_refined0.3160.23303
X-RAY DIFFRACTIONr_xyhbond_nbd_refined0.130.2467
X-RAY DIFFRACTIONr_metal_ion_refined0.3780.29
X-RAY DIFFRACTIONr_symmetry_vdw_refined0.2660.2117
X-RAY DIFFRACTIONr_symmetry_hbond_refined0.1860.228
X-RAY DIFFRACTIONr_symmetry_metal_ion_refined0.1150.24
X-RAY DIFFRACTIONr_mcbond_it2.59422935
X-RAY DIFFRACTIONr_mcangle_it3.72934788
X-RAY DIFFRACTIONr_scbond_it3.18522059
X-RAY DIFFRACTIONr_scangle_it4.44931954
Refine LS restraints NCS

Dom-ID: 1 / Refine-ID: X-RAY DIFFRACTION

Ens-IDAuth asym-IDNumberTypeRms dev position (Å)Weight position
1A672TIGHT POSITIONAL0.020.05
1A25LOOSE POSITIONAL0.675
1A672TIGHT THERMAL0.371
1A25LOOSE THERMAL1.1320
2B677TIGHT POSITIONAL0.030.05
2B34LOOSE POSITIONAL0.735
2B677TIGHT THERMAL0.331
2B34LOOSE THERMAL120
3C608TIGHT POSITIONAL0.020.05
3C608TIGHT THERMAL0.321
LS refinement shellResolution: 1.779→1.825 Å / Total num. of bins used: 20
RfactorNum. reflection% reflection
Rfree0.324 227 -
Rwork0.274 4508 -
obs-4735 94 %
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)
12.8368-1.51870.02984.14510.31983.4911-0.1114-0.03570.0298-0.099-0.0160.0346-0.0301-0.0570.1274-0.24710.0501-0.0379-0.1961-0.0729-0.231314.665-18.057-21.123
23.077-0.8261-1.01884.6283-0.4365.4606-0.1201-0.1714-0.47550.313-0.03890.26440.4399-0.05970.159-0.1910.0250.004-0.1711-0.0424-0.129114.586-27.1324.801
33.45140.74150.962110.14112.78714.60370.0211-0.3561-0.15530.70850.4267-0.6350.33090.3694-0.4478-0.14120.0302-0.0143-0.1317-0.0535-0.122812.868-10.61125.539
42.54911.36950.20194.22360.35023.449-0.10440.0176-0.05320.1095-0.01420.01940.0385-0.05150.1186-0.2463-0.04660.0372-0.1907-0.0704-0.233114.70817.5921.332
52.9660.80971.13044.3397-0.29815.8082-0.150.14910.4751-0.2958-0.01210.2751-0.4664-0.05510.1621-0.1888-0.0226-0.0111-0.1796-0.0427-0.125814.50126.972-4.634
63.5038-0.8589-1.0810.16192.68184.50040.02360.35040.1249-0.78630.392-0.5894-0.33360.3574-0.4156-0.1282-0.03240.0054-0.1329-0.0465-0.125912.88110.242-25.637
Refinement TLS group

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

IDRefine TLS-IDAuth asym-IDLabel asym-IDAuth seq-IDLabel seq-ID
11AA149 - 24610 - 107
22BB144 - 2465 - 107
33CC148 - 2469 - 107
44DD149 - 24610 - 107
55EE144 - 2465 - 107
66FF148 - 2469 - 107

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