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- PDB-2pft: The Crystal Structure of Mouse Exo70 Reveals Unique Features of t... -

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Basic information

Entry
Database: PDB / ID: 2pft
TitleThe Crystal Structure of Mouse Exo70 Reveals Unique Features of the Mammalian Exocyst
ComponentsExocytosis Protein
KeywordsENDOCYTOSIS/EXOCYTOSIS / helix-turn-helix / ENDOCYTOSIS-EXOCYTOSIS COMPLEX
Function / homology
Function and homology information


Insulin processing / VxPx cargo-targeting to cilium / regulation of entry of bacterium into host cell / exocyst / growth cone membrane / Flemming body / microtubule organizing center / exocytosis / centriolar satellite / protein transport ...Insulin processing / VxPx cargo-targeting to cilium / regulation of entry of bacterium into host cell / exocyst / growth cone membrane / Flemming body / microtubule organizing center / exocytosis / centriolar satellite / protein transport / plasma membrane / cytosol
Similarity search - Function
Exocyst complex component Exo70 / Exocyst complex component Exo70 / Exo70 exocyst complex subunit C-terminal / Cullin repeat-like-containing domain superfamily / Monooxygenase / Up-down Bundle / Mainly Alpha
Similarity search - Domain/homology
Exocyst complex component 7 / Exocyst complex component 7
Similarity search - Component
Biological speciesMus musculus (house mouse)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MAD / Resolution: 2.25 Å
AuthorsMoore, B.A. / Xu, Z.
CitationJournal: J.Mol.Biol. / Year: 2007
Title: The crystal structure of mouse Exo70 reveals unique features of the mammalian exocyst.
Authors: Moore, B.A. / Robinson, H.H. / Xu, Z.
History
DepositionApr 5, 2007Deposition site: RCSB / Processing site: RCSB
Revision 1.0May 22, 2007Provider: repository / Type: Initial release
Revision 1.1Dec 17, 2007Group: Version format compliance
Revision 1.2Jul 13, 2011Group: Version format compliance
Revision 1.3Nov 16, 2011Group: Atomic model
Revision 1.4Oct 18, 2017Group: Refinement description / Category: software
Revision 1.5Feb 21, 2024Group: Data collection / Database references
Category: chem_comp_atom / chem_comp_bond ...chem_comp_atom / chem_comp_bond / database_2 / struct_ref_seq_dif
Item: _database_2.pdbx_DOI / _database_2.pdbx_database_accession / _struct_ref_seq_dif.details

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: Exocytosis Protein


Theoretical massNumber of molelcules
Total (without water)65,5551
Polymers65,5551
Non-polymers00
Water3,063170
1


  • Idetical with deposited unit
  • defined by author
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Unit cell
Length a, b, c (Å)61.520, 61.520, 294.730
Angle α, β, γ (deg.)90.000, 90.000, 120.000
Int Tables number154
Space group name H-MP3221
Components on special symmetry positions
IDModelComponents
11A-812-

HOH

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Components

#1: Protein Exocytosis Protein /


Mass: 65555.477 Da / Num. of mol.: 1 / Fragment: Residues 85-653
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Mus musculus (house mouse) / Gene: Exoc7 / Plasmid: pSJ7 / Species (production host): Escherichia coli / Production host: Escherichia coli BL21(DE3) (bacteria) / Strain (production host): BL21(DE3) / References: UniProt: Q3TIP4, UniProt: O35250*PLUS
#2: Water ChemComp-HOH / water / Water


Mass: 18.015 Da / Num. of mol.: 170 / 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: 2.46 Å3/Da / Density % sol: 49.9 %
Crystal growTemperature: 293.15 K / Method: vapor diffusion, sitting drop / pH: 7.5
Details: Precipitant in drop: 10% ethylene glycol, 5% PEG 8000, 0.01M magnesium chloride, 0.1M HEPES Precipitant in well: 10% ethylene glycol, 8% PEG 8000, 0.1M HEPES, pH 7.5, VAPOR DIFFUSION, ...Details: Precipitant in drop: 10% ethylene glycol, 5% PEG 8000, 0.01M magnesium chloride, 0.1M HEPES Precipitant in well: 10% ethylene glycol, 8% PEG 8000, 0.1M HEPES, pH 7.5, VAPOR DIFFUSION, SITTING DROP, temperature 293.15K

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Data collection

Diffraction
IDMean temperature (K)Crystal-ID
1771
21
31
Diffraction sourceSource: SYNCHROTRON / Site: APS / Beamline: 23-ID-D / Wavelength: 0.97926,0.97942,0.95660
DetectorType: MARMOSAIC 300 mm CCD / Detector: CCD / Date: Jul 19, 2006 / Details: Adjustable focusing mirrors in K-B geometry
RadiationMonochromator: Si(111) Double Crystal / Protocol: MAD / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelength
IDWavelength (Å)Relative weight
10.979261
20.979421
30.95661
Reflection

D res high: 2.5 Å / D res low: 50 Å

Redundancy (%)IDAv σ(I) over netINumberRmerge(I) obsΧ2Num. obs% possible obs
6.2115.41441400.0792.762332098.6
5.7215.31313130.0692.042300397.1
5.4314.71194440.0691.912228493.4
Diffraction reflection shell
Highest resolution (Å)Lowest resolution (Å)% possible obs (%)IDRmerge(I) obsChi squaredRedundancy
5.38509810.076.8615.8
4.275.3899.310.0724.6826.1
3.734.2710010.0693.3826.4
3.393.7310010.0813.0496.4
3.153.3910010.1012.5156.6
2.963.1510010.1151.7126.7
2.822.9610010.141.386.6
2.692.8210010.1791.2276.4
2.592.6999.610.2531.276
2.52.5989.210.2970.9824.6
5.385098.120.0523.5665.6
4.275.3899.420.0622.9826
3.734.2799.920.0632.4396.2
3.393.7310020.0792.46.3
3.153.3999.920.1051.9396.4
2.963.1599.920.1351.4316.5
2.822.9610020.1821.2446.3
2.692.8299.520.2361.1265.6
2.592.6996.520.3181.1044.5
2.52.5977.520.3651.1173
5.385097.930.052.8545.5
4.275.3899.630.0622.7045.9
3.734.2799.830.0672.3716
3.393.7399.630.0892.2656.2
3.153.3999.830.131.8146.3
2.963.1599.930.1761.3046.2
2.822.9699.730.2291.095.6
2.692.8294.130.2951.0154.6
2.592.6983.130.3860.9913.3
2.52.5959.230.4230.9722.3
ReflectionResolution: 2.25→46.84 Å / Num. all: 31423 / Num. obs: 30334 / % possible obs: 93.7 % / Observed criterion σ(F): 0 / Redundancy: 4.2 % / Biso Wilson estimate: 40.4 Å2 / Limit h max: 24 / Limit h min: 0 / Limit k max: 24 / Limit k min: 0 / Limit l max: 133 / Limit l min: 0 / Observed criterion F max: 127192.55 / Observed criterion F min: 0.32 / Rmerge(I) obs: 0.054 / Χ2: 1.26 / Net I/σ(I): 15.8
Reflection shellResolution: 2.25→2.33 Å / Redundancy: 3.1 % / Rmerge(I) obs: 0.416 / Num. unique all: 2448 / Χ2: 1.093 / % possible all: 77.7

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Phasing

PhasingMethod: MAD
Phasing MAD set
IDR cullis acentricR cullis centricHighest resolution (Å)Lowest resolution (Å)Reflection acentricReflection centric
ISO_1002.546.9194713780
ISO_20.3820.3452.546.9191343700
ISO_30.5950.572.546.9183453614
ANO_10.51402.546.9190690
ANO_20.69302.546.9179810
ANO_30.79602.546.9165810
Phasing MAD set shell
IDResolution (Å)R cullis acentricR cullis centricReflection acentricReflection centric
ISO_110.89-46.900159157
ISO_17.81-10.8900339187
ISO_16.4-7.8100489194
ISO_15.56-6.400578195
ISO_14.98-5.5600687189
ISO_14.55-4.9800752180
ISO_14.22-4.5500823200
ISO_13.94-4.2200889192
ISO_13.72-3.9400970201
ISO_13.53-3.72001021188
ISO_13.37-3.53001083190
ISO_13.22-3.37001108197
ISO_13.1-3.22001179202
ISO_12.99-3.1001268179
ISO_12.89-2.99001258207
ISO_12.79-2.89001322198
ISO_12.71-2.79001386191
ISO_12.64-2.71001414195
ISO_12.57-2.64001455175
ISO_12.5-2.57001291163
ANO_110.89-46.90.32901490
ANO_17.81-10.890.36903310
ANO_16.4-7.810.29904880
ANO_15.56-6.40.29505740
ANO_14.98-5.560.36806780
ANO_14.55-4.980.4707390
ANO_14.22-4.550.43208080
ANO_13.94-4.220.45508750
ANO_13.72-3.940.47509600
ANO_13.53-3.720.546010040
ANO_13.37-3.530.553010770
ANO_13.22-3.370.567011030
ANO_13.1-3.220.59011750
ANO_12.99-3.10.657012670
ANO_12.89-2.990.699012550
ANO_12.79-2.890.768013200
ANO_12.71-2.790.806013810
ANO_12.64-2.710.937013940
ANO_12.57-2.640.925013960
ANO_12.5-2.570.952010950
ISO_210.89-46.90.3310.365155155
ISO_27.81-10.890.3420.251337185
ISO_26.4-7.810.1810.17489193
ISO_25.56-6.40.1720.164578195
ISO_24.98-5.560.2030.188686189
ISO_24.55-4.980.2410.281751178
ISO_24.22-4.550.2420.272823197
ISO_23.94-4.220.2490.308889192
ISO_23.72-3.940.270.311970201
ISO_23.53-3.720.2740.3251021188
ISO_23.37-3.530.3280.3891082190
ISO_23.22-3.370.3710.451105197
ISO_23.1-3.220.4640.5461178201
ISO_22.99-3.10.5210.7061268178
ISO_22.89-2.990.6120.7171256207
ISO_22.79-2.890.710.7891320197
ISO_22.71-2.790.7720.871378187
ISO_22.64-2.710.4380.8731398185
ISO_22.57-2.640.8250.8971378160
ISO_22.5-2.570.8580.9081072125
ANO_210.89-46.90.47101490
ANO_27.81-10.890.48903310
ANO_26.4-7.810.36304870
ANO_25.56-6.40.41105760
ANO_24.98-5.560.52806780
ANO_24.55-4.980.64407310
ANO_24.22-4.550.61708020
ANO_23.94-4.220.64708600
ANO_23.72-3.940.6609500
ANO_23.53-3.720.74309960
ANO_23.37-3.530.746010640
ANO_23.22-3.370.786010880
ANO_23.1-3.220.793011670
ANO_22.99-3.10.865012590
ANO_22.89-2.990.881012500
ANO_22.79-2.890.92013030
ANO_22.71-2.790.931012950
ANO_22.64-2.710.974012280
ANO_22.57-2.640.978010960
ANO_22.5-2.570.98306710
ISO_310.89-46.90.5120.528155152
ISO_37.81-10.890.5060.546338185
ISO_36.4-7.810.5090.474489194
ISO_35.56-6.40.5380.566577195
ISO_34.98-5.560.5130.545686187
ISO_34.55-4.980.4920.492752178
ISO_34.22-4.550.5140.521823199
ISO_33.94-4.220.5040.567889192
ISO_33.72-3.940.5410.585967200
ISO_33.53-3.720.5180.5541019187
ISO_33.37-3.530.5810.6021075189
ISO_33.22-3.370.6290.6851106197
ISO_33.1-3.220.6990.7121178202
ISO_32.99-3.10.7530.8511266179
ISO_32.89-2.990.7810.8751255204
ISO_32.79-2.890.8450.931317191
ISO_32.71-2.790.8640.961308173
ISO_32.64-2.710.8760.9591257163
ISO_32.57-2.640.8650.9211135151
ISO_32.5-2.570.8941.00575396
ANO_310.89-46.90.50501440
ANO_37.81-10.890.61903300
ANO_36.4-7.810.5104860
ANO_35.56-6.40.51305710
ANO_34.98-5.560.66906760
ANO_34.55-4.980.77607310
ANO_34.22-4.550.74607910
ANO_33.94-4.220.7608510
ANO_33.72-3.940.80209320
ANO_33.53-3.720.85109850
ANO_33.37-3.530.856010470
ANO_33.22-3.370.88010880
ANO_33.1-3.220.904011700
ANO_32.99-3.10.943012570
ANO_32.89-2.990.97012300
ANO_32.79-2.890.976012360
ANO_32.71-2.790.969010960
ANO_32.64-2.710.97808840
ANO_32.57-2.640.99106610
ANO_32.5-2.570.99204150
Phasing MAD set site
IDCartn x (Å)Cartn y (Å)Cartn z (Å)Atom type symbolB isoOccupancy
1-31.791-22.349-0.665SE60.580.91
2-7.811-42.177-10.244SE56.80.94
316.393-31.502-14.809SE57.670.94
4-35.063-41.92-3.793SE55.51.11
5-10.793-23.465-42.746SE71.711.05
6-40.086-28.025-21.986SE86.261.08
7-49.165-2.637-28.712SE79.70.88
8-34.492-21.508-23.78SE79.510.89
9-7.261-11.69-30.628SE71.380.58
10-5.524-27.151-7.297SE100.330.84

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Processing

Software
NameVersionClassificationNB
DENZOdata reduction
SCALEPACKdata scaling
SHARPphasing
SOLOMONphasing
CNS1.2refinement
PDB_EXTRACT2data extraction
Blu-Ice(Custom)data collection
HKL-2000data reduction
HKL-2000data scaling
RefinementMethod to determine structure: MAD / Resolution: 2.25→46.84 Å / Rfactor Rfree error: 0.008 / Occupancy max: 1 / Occupancy min: 1 / Data cutoff high absF: 127010.656 / Data cutoff low absF: 0 / Isotropic thermal model: RESTRAINED / Cross valid method: THROUGHOUT / σ(F): 0 / Details: BULK SOLVENT MODEL USED
RfactorNum. reflection% reflectionSelection details
Rfree0.285 1435 5 %RANDOM
Rwork0.235 ---
all-32008 --
obs-28804 90 %-
Solvent computationSolvent model: FLAT MODEL / Bsol: 57.814 Å2 / ksol: 0.35 e/Å3
Displacement parametersBiso max: 98.02 Å2 / Biso mean: 54.8 Å2 / Biso min: 29.02 Å2
Baniso -1Baniso -2Baniso -3
1-2.06 Å20 Å20 Å2
2--2.06 Å20 Å2
3----4.11 Å2
Refine analyze
FreeObs
Luzzati coordinate error0.4 Å0.32 Å
Luzzati d res low-5 Å
Luzzati sigma a0.38 Å0.28 Å
Luzzati d res high-2.25
Refinement stepCycle: LAST / Resolution: 2.25→46.84 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms4098 0 0 170 4268
Refine LS restraints
Refine-IDTypeDev idealDev ideal target
X-RAY DIFFRACTIONx_bond_d0.006
X-RAY DIFFRACTIONx_angle_deg1.1
X-RAY DIFFRACTIONx_torsion_deg17.9
X-RAY DIFFRACTIONx_torsion_impr_deg0.85
X-RAY DIFFRACTIONx_mcbond_it1.431.5
X-RAY DIFFRACTIONx_mcangle_it2.362
X-RAY DIFFRACTIONx_scbond_it2.172
X-RAY DIFFRACTIONx_scangle_it3.232.5
LS refinement shell

Refine-ID: X-RAY DIFFRACTION

Resolution (Å)Rfactor RfreeNum. reflection Rfree% reflection Rfree (%)Rfactor RworkNum. reflection RworkRfactor Rfree errorNum. reflection allNum. reflection obs% reflection obs (%)
2.25-2.350.3781344.80.3126430.0333933277770.6
2.35-2.480.36215950.28329920.0293936315180.1
2.48-2.630.3251985.80.28331900.0233907338886.7
2.63-2.830.3141915.30.25934330.0233936362492
2.83-3.120.3461624.30.27636100.0273992377294.5
3.12-3.570.3141995.20.25436610.0224002386096.5
3.57-4.50.2381934.80.21237910.0174044398498.5
4.5-46.830.2741994.70.21940490.0194287424899.1
Xplor file
Refine-IDSerial noParam fileTopol file
X-RAY DIFFRACTION1protein_rep.paramprotein.top
X-RAY DIFFRACTION2water_rep.param

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