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- PDB-9m73: Crystal structure of MBP-fused BIL1/BZR1 (21-104) in complex with... -

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Basic information

Entry
Database: PDB / ID: 9m73
TitleCrystal structure of MBP-fused BIL1/BZR1 (21-104) in complex with double-stranded DNA contaning CACATATGTG
Components
  • E-box(CATATG)-containing DNA
  • Maltodextrin-binding protein,Protein BRASSINAZOLE-RESISTANT 1
KeywordsDNA BINDING PROTEIN / Plant-specific / DNA-biding protein / Transcription factor / BZR
Function / homology
Function and homology information


plant ovule development / seed development / brassinosteroid mediated signaling pathway / cell envelope / carbohydrate transmembrane transporter activity / maltose binding / maltose transport / maltodextrin transmembrane transport / ATP-binding cassette (ABC) transporter complex, substrate-binding subunit-containing / periplasmic space ...plant ovule development / seed development / brassinosteroid mediated signaling pathway / cell envelope / carbohydrate transmembrane transporter activity / maltose binding / maltose transport / maltodextrin transmembrane transport / ATP-binding cassette (ABC) transporter complex, substrate-binding subunit-containing / periplasmic space / DNA-binding transcription factor activity / negative regulation of DNA-templated transcription / DNA-templated transcription / regulation of DNA-templated transcription / DNA binding / metal ion binding / nucleus / cytosol
Similarity search - Function
BZR family / BES1/BZR1 plant transcription factor, N-terminal / BES1/BZR1 plant transcription factor, N-terminal / 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 / ACP-like superfamily / Carrier protein (CP) domain profile. / Phosphopantetheine binding ACP domain
Similarity search - Domain/homology
alpha-maltose / DNA / DNA (> 10) / Maltodextrin-binding protein / Protein BRASSINAZOLE-RESISTANT 1
Similarity search - Component
Biological speciesArabidopsis thaliana (thale cress)
synthetic construct (others)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 2.31 Å
AuthorsShohei, N. / Masaru, T. / Takuya, M.
Funding support Japan, 6items
OrganizationGrant numberCountry
Ministry of Education, Culture, Sports, Science and Technology (Japan)JP17H05835 Japan
Ministry of Education, Culture, Sports, Science and Technology (Japan)JP19H04855 Japan
Ministry of Education, Culture, Sports, Science and Technology (Japan)JP19K23658 Japan
Ministry of Education, Culture, Sports, Science and Technology (Japan)JP21H02114 Japan
Japan Agency for Medical Research and Development (AMED)JP20am0101077 Japan
Japan Agency for Medical Research and Development (AMED)JP24ama121010 Japan
CitationJournal: Febs Lett. / Year: 2025
Title: Single cis-elements in brassinosteroid-induced upregulated genes are insufficient to recruit both redox states of the BIL1/BZR1 DNA-binding domain.
Authors: Nosaki, S. / Ohtsuka, M. / Nakano, T. / Tanokura, M. / Miyakawa, T.
History
DepositionMar 9, 2025Deposition site: PDBJ / Processing site: PDBJ
Revision 1.0Sep 10, 2025Provider: repository / Type: Initial release

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
C: Maltodextrin-binding protein,Protein BRASSINAZOLE-RESISTANT 1
D: Maltodextrin-binding protein,Protein BRASSINAZOLE-RESISTANT 1
G: E-box(CATATG)-containing DNA
H: E-box(CATATG)-containing DNA
A: Maltodextrin-binding protein,Protein BRASSINAZOLE-RESISTANT 1
B: Maltodextrin-binding protein,Protein BRASSINAZOLE-RESISTANT 1
E: E-box(CATATG)-containing DNA
F: E-box(CATATG)-containing DNA
hetero molecules


Theoretical massNumber of molelcules
Total (without water)219,32517
Polymers217,6468
Non-polymers1,6809
Water2,864159
1
C: Maltodextrin-binding protein,Protein BRASSINAZOLE-RESISTANT 1
D: Maltodextrin-binding protein,Protein BRASSINAZOLE-RESISTANT 1
G: E-box(CATATG)-containing DNA
H: E-box(CATATG)-containing DNA
hetero molecules


Theoretical massNumber of molelcules
Total (without water)109,5707
Polymers108,8234
Non-polymers7473
Water724
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area9430 Å2
ΔGint-19 kcal/mol
Surface area41300 Å2
MethodPISA
2
A: Maltodextrin-binding protein,Protein BRASSINAZOLE-RESISTANT 1
B: Maltodextrin-binding protein,Protein BRASSINAZOLE-RESISTANT 1
E: E-box(CATATG)-containing DNA
F: E-box(CATATG)-containing DNA
hetero molecules


Theoretical massNumber of molelcules
Total (without water)109,75610
Polymers108,8234
Non-polymers9336
Water724
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area9650 Å2
ΔGint-2 kcal/mol
Surface area41460 Å2
MethodPISA
Unit cell
Length a, b, c (Å)102.848, 92.602, 111.767
Angle α, β, γ (deg.)90.000, 100.323, 90.000
Int Tables number4
Space group name H-MP1211
Space group name HallP2yb
Symmetry operation#1: x,y,z
#2: -x,y+1/2,-z

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Components

#1: Protein
Maltodextrin-binding protein,Protein BRASSINAZOLE-RESISTANT 1 / Protein BIN2 SUBSTRATE 2


Mass: 49819.445 Da / Num. of mol.: 4
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Arabidopsis thaliana (thale cress) / Gene: malE, JW3994, BZR1, BIS2, At1g75080, F9E10.7 / Production host: Escherichia coli BL21(DE3) (bacteria) / References: UniProt: A0A4P1LXE0, UniProt: Q8S307
#2: DNA chain
E-box(CATATG)-containing DNA


Mass: 4592.023 Da / Num. of mol.: 4
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) synthetic construct (others) / Production host: synthetic construct (others)
#3: Polysaccharide
alpha-D-glucopyranose-(1-4)-alpha-D-glucopyranose


Type: oligosaccharide, Oligosaccharide / Class: Nutrient / Mass: 342.297 Da / Num. of mol.: 4
Source method: isolated from a genetically manipulated source
Details: oligosaccharide / References: alpha-maltose
DescriptorTypeProgram
DGlcpa1-4DGlcpa1-ROHGlycam Condensed SequenceGMML 1.0
WURCS=2.0/1,2,1/[a2122h-1a_1-5]/1-1/a4-b1WURCSPDB2Glycan 1.1.0
[][a-D-Glcp]{[(4+1)][a-D-Glcp]{}}LINUCSPDB-CARE
#4: Chemical
ChemComp-EDO / 1,2-ETHANEDIOL / ETHYLENE GLYCOL


Mass: 62.068 Da / Num. of mol.: 5 / Source method: obtained synthetically / Formula: C2H6O2
#5: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 159 / Source method: isolated from a natural source / Formula: H2O
Has ligand of interestN
Has protein modificationN

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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: 2.56 Å3/Da / Density % sol: 51.99 %
Crystal growTemperature: 293 K / Method: vapor diffusion, sitting drop / pH: 6.5
Details: 50 mM sodium cacodylate pH 6.5, 200 mM potassium chloride, 10 mM magnesium chloride and 10% (w/v) polyethylene glycol (PEG) 4000

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Data collection

DiffractionMean temperature: 95 K / Ambient temp details: DECTRIS PILATUS 2M-F / Serial crystal experiment: N
Diffraction sourceSource: SYNCHROTRON / Site: Photon Factory / Beamline: AR-NE3A / Wavelength: 1 Å
DetectorType: DECTRIS PILATUS 2M-F / Detector: PIXEL / Date: Apr 19, 2017
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 1 Å / Relative weight: 1
ReflectionResolution: 2.31→43.62 Å / Num. obs: 90599 / % possible obs: 100 % / Redundancy: 7 % / Biso Wilson estimate: 42.68 Å2 / CC1/2: 0.999 / Rpim(I) all: 0.047 / Net I/σ(I): 16
Reflection shellResolution: 2.31→2.35 Å / Redundancy: 6.9 % / Mean I/σ(I) obs: 2.1 / Num. unique obs: 4450 / CC1/2: 0.746 / Rpim(I) all: 0.557 / % possible all: 100

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Processing

Software
NameVersionClassification
PHENIX1.18_3855refinement
XDSdata reduction
Aimlessdata scaling
PHASERphasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT / Resolution: 2.31→43.62 Å / SU ML: 0.2828 / Cross valid method: FREE R-VALUE / σ(F): 1.34 / Phase error: 27.2744
Stereochemistry target values: GeoStd + Monomer Library + CDL v1.2
RfactorNum. reflection% reflection
Rfree0.2451 4565 5.04 %
Rwork0.2054 85940 -
obs0.2074 90505 99.88 %
Solvent computationShrinkage radii: 0.9 Å / VDW probe radii: 1.11 Å / Solvent model: FLAT BULK SOLVENT MODEL
Displacement parametersBiso mean: 59.16 Å2
Refinement stepCycle: LAST / Resolution: 2.31→43.62 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms13584 1220 112 159 15075
Refine LS restraints
Refine-IDTypeDev idealNumber
X-RAY DIFFRACTIONf_bond_d0.005815379
X-RAY DIFFRACTIONf_angle_d0.691621116
X-RAY DIFFRACTIONf_chiral_restr0.04532305
X-RAY DIFFRACTIONf_plane_restr0.00412525
X-RAY DIFFRACTIONf_dihedral_angle_d25.26855749
LS refinement shell
Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkRefine-ID% reflection obs (%)
2.31-2.340.34071260.27652869X-RAY DIFFRACTION99.9
2.34-2.360.34561390.27832841X-RAY DIFFRACTION100
2.36-2.390.31791420.25982889X-RAY DIFFRACTION100
2.39-2.420.29791580.25452843X-RAY DIFFRACTION99.93
2.42-2.450.28821520.25842860X-RAY DIFFRACTION100
2.45-2.490.31841540.25812829X-RAY DIFFRACTION100
2.49-2.520.30661410.24892861X-RAY DIFFRACTION100
2.52-2.560.30981630.24862860X-RAY DIFFRACTION100
2.56-2.60.26931870.23482822X-RAY DIFFRACTION100
2.6-2.640.28951570.23172866X-RAY DIFFRACTION99.97
2.64-2.690.26721580.23132821X-RAY DIFFRACTION100
2.69-2.740.27881410.23792874X-RAY DIFFRACTION99.97
2.74-2.790.29821530.24792870X-RAY DIFFRACTION99.93
2.79-2.850.37421510.26172847X-RAY DIFFRACTION100
2.85-2.910.26961510.25832844X-RAY DIFFRACTION100
2.91-2.980.33921400.24472872X-RAY DIFFRACTION100
2.98-3.050.28031640.23932868X-RAY DIFFRACTION100
3.05-3.130.2911330.24072874X-RAY DIFFRACTION99.9
3.14-3.230.29481780.24682853X-RAY DIFFRACTION99.9
3.23-3.330.29811460.23262841X-RAY DIFFRACTION99.9
3.33-3.450.23711660.21792872X-RAY DIFFRACTION99.97
3.45-3.590.23891400.20542867X-RAY DIFFRACTION100
3.59-3.750.26561510.20432869X-RAY DIFFRACTION100
3.75-3.950.22691210.18812924X-RAY DIFFRACTION99.93
3.95-4.20.22121540.16652861X-RAY DIFFRACTION99.97
4.2-4.520.18521680.15812858X-RAY DIFFRACTION99.9
4.52-4.970.18761570.15192886X-RAY DIFFRACTION99.93
4.98-5.690.20821570.17522890X-RAY DIFFRACTION99.77
5.69-7.170.20711750.19022884X-RAY DIFFRACTION99.8
7.17-43.620.16851420.1662925X-RAY DIFFRACTION97.8
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)
15.14114713681-0.625460863467-2.0220914692.909269068020.4201112025363.155236228110.184499202785-0.2569744814020.51245371863-0.002704538733170.0325726968563-0.187471724641-0.3271254895820.306979699615-0.1780406009950.371867652302-0.07414342716730.04778069050870.2602248436550.01367207027650.284414990616-2.5709070165914.233053860711.4420975675
24.90462819306-1.627877876190.8333512090344.75563783394-1.248887012663.48353176001-0.00383965421563-0.370195606995-0.7544113739850.4667731469670.104413706559-0.0493530011970.923850050888-0.19506701946-0.08659831376660.611085039695-0.0903866330332-0.03688504217140.292591829024-0.008774151047810.418142442404-6.38720904937-37.013372728727.6808195299
31.36187315310.287782156283-0.1012855761253.128453247720.6145414547722.76117702664-0.193783883896-0.1888723881840.06648493245110.5243318973590.639658632081-0.730331560949-0.2935647364120.573043779139-0.4185190059530.5677780861310.15516301954-0.05710245268770.718152828454-0.2493401870710.464399986349-8.6705984338113.638182421745.3018114572
43.819673191551.471967979810.5410462567793.474834179391.41634762572.988055955-0.03508746314030.226701562466-0.630401765573-0.3979062991720.1249153900310.04417965773130.807819947543-0.20926289731-0.06791186333410.649586549783-0.0662752331539-0.06344446177460.2543161355120.04135664635690.444513139242-1.96712018335-36.885564394425.6342343889
51.86870641803-0.4379341174990.6476270756722.92803048116-1.85824921819.333238106180.384495833761-0.482918627326-0.2102421985160.5376357532270.189774200608-0.0206465964214-0.116069358274-0.400839409597-0.5492863098690.421408065627-0.15075162704-0.007832020981970.4031113959020.006038438845280.338354140716-5.88963661978-23.870235858828.6158245743
63.72727380132-0.532827416541-3.471421000093.686247223971.87710485633.75299373060.532308714170.259168186257-0.03544695091580.05180075242220.0261569404416-0.263768924586-0.664618820941-0.117214974896-0.5242834427220.459599213073-0.0315361263088-0.04794187587940.3614412740130.01508182743440.297286545768-2.91458474868-23.823987206724.7228946266
74.560856684060.367641221678-1.375085914251.26734581036-0.2624193445571.697149154420.081672810349-0.559268388799-0.04226686825560.121117835251-0.13927642741-0.1024615016040.03913997589430.6402031035670.05287480069580.3311271638770.04291127295120.006103438782340.51928613380.01358151599110.16085852121639.8584413002-13.144879637643.8982211553
81.16786749976-2.006052600841.02956230027.52627661793-6.034937419235.24854664949-0.0213707757904-0.1045696433050.08091444160180.09051026972140.150166685760.486525677708-0.234148048953-0.541111877998-0.1300320708370.660257957635-0.0606060203511-0.06129778014010.700989210304-0.245219828720.4543160353445.794136864729.854513667829.9040777436
93.44205450061-0.17912586478-0.3397687552391.3562606552-0.2179083272781.80342175351-0.05488601241360.0853400552435-0.0450109647794-0.05920927476420.03722956296110.1277048513990.0434940739175-0.4153469002040.02053226315540.293779041624-0.04722614172020.01080311471070.307958591615-0.006350831103190.14925760713953.2703451938-15.130963779211.0775701908
100.8188684358830.752654321430.09389166262335.885485110142.77659490711.30584253488-0.107428214853-0.291956640064-0.03158062436540.7617449846060.272428258962-0.3726876003790.2308104010650.405368840612-0.1864706864420.7298554649020.175559742299-0.1098474875860.7007380978370.09286051426270.43990743934947.353126850129.902592683424.7359314966
110.211034005470.0543557570250.9167129448321.313771377412.169222649166.718158195040.316800973761-0.440429312171-0.0670862531630.362188828320.2185033851670.1174015951832.55328634522-0.712393419471-0.4762732971191.077842071630.00525712560095-0.06930301340961.18326487550.06772154358320.41664868694246.988122925123.656489172229.809453598
120.7251543370160.723903819-1.125419201561.0722025563-2.698415604538.584747267960.207194648942-0.715828092751-0.05541843189960.2407512542760.237999408348-0.2059217671312.286097173491.26331622197-0.4186428735641.220802911360.212989727205-0.004383825830961.202448457490.02953030992730.44594845780345.376201271323.539239123725.5664565524
Refinement TLS group

Refine-ID: X-RAY DIFFRACTION

IDRefine TLS-IDSelection detailsAuth asym-IDLabel asym-IDAuth seq-IDLabel seq-ID
11chain 'C' and (resid -347 through 26 )CA-347 - 261 - 374
22chain 'C' and (resid 27 through 88 )CA27 - 88375 - 436
33chain 'D' and (resid -347 through 26 )DD-347 - 261 - 374
44chain 'D' and (resid 27 through 88 )DD27 - 88375 - 436
55chain 'G' and (resid -3 through 11 )GF-3 - 11
66chain 'H' and (resid -3 through 11 )HG-3 - 11
77chain 'A' and (resid -348 through 3 )AH-348 - 31 - 352
88chain 'A' and (resid 4 through 88 )AH4 - 88353 - 437
99chain 'B' and (resid -347 through 4 )BK-347 - 41 - 352
1010chain 'B' and (resid 5 through 88 )BK5 - 88353 - 436
1111chain 'E' and (resid -3 through 11 )EP-3 - 11
1212chain 'F' and (resid -3 through 11 )FQ-3 - 11

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