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- PDB-3lrd: Self-assembly of spider silk proteins is controlled by a pH-sensi... -

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Basic information

Entry
Database: PDB / ID: 3lrd
TitleSelf-assembly of spider silk proteins is controlled by a pH-sensitive relay
ComponentsMajor ampullate spidroin 1
KeywordsSTRUCTURAL PROTEIN / dragline spider silk / self-assembly / pH-dependence / NT D40N/E84Q double-mutant
Function / homology
Function and homology information


identical protein binding
Similarity search - Function
Spidroin, N-terminal domain / Spidroin, N-terminal / Major ampullate spidroin 1, spider silk protein 1, N-term / Spidroin, N-terminal domain superfamily / Enzyme I; Chain A, domain 2 / Orthogonal Bundle / Mainly Alpha
Similarity search - Domain/homology
TRIETHYLENE GLYCOL / Major ampullate spidroin 1
Similarity search - Component
Biological speciesEuprosthenops australis (spider)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 2.15 Å
AuthorsAskarieh, G. / Hedhammar, H. / Nordling, K. / Rising, A. / Johansson, J. / Knight, S.D. / Saenz, A. / Casals, C.
CitationJournal: Nature / Year: 2010
Title: Self-assembly of spider silk proteins is controlled by a pH-sensitive relay.
Authors: Askarieh, G. / Hedhammar, M. / Nordling, K. / Saenz, A. / Casals, C. / Rising, A. / Johansson, J. / Knight, S.D.
History
DepositionFeb 11, 2010Deposition site: RCSB / Processing site: RCSB
Revision 1.0May 12, 2010Provider: repository / Type: Initial release
Revision 1.1Jul 13, 2011Group: Version format compliance
Revision 1.2Oct 13, 2021Group: Database references / Derived calculations / Category: database_2 / 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_ref_seq_dif.details / _struct_site.pdbx_auth_asym_id / _struct_site.pdbx_auth_comp_id / _struct_site.pdbx_auth_seq_id
Revision 1.3Feb 21, 2024Group: Data collection / Category: chem_comp_atom / chem_comp_bond
Revision 1.4Apr 3, 2024Group: Refinement description / Category: pdbx_initial_refinement_model

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: Major ampullate spidroin 1
B: Major ampullate spidroin 1
hetero molecules


Theoretical massNumber of molelcules
Total (without water)28,6385
Polymers28,3632
Non-polymers2743
Water99155
1


  • Idetical with deposited unit
  • defined by author&software
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area3420 Å2
ΔGint-14 kcal/mol
Surface area10130 Å2
MethodPISA
Unit cell
Length a, b, c (Å)68.383, 68.383, 97.781
Angle α, β, γ (deg.)90.00, 90.00, 120.00
Int Tables number154
Space group name H-MP3221
Components on special symmetry positions
IDModelComponents
11B-155-

HOH

21B-175-

HOH

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Components

#1: Protein Major ampullate spidroin 1


Mass: 14181.743 Da / Num. of mol.: 2 / Fragment: N-terminal domain / Mutation: D40N, E84Q
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Euprosthenops australis (spider) / Gene: MaSp1 / Production host: Escherichia coli (E. coli) / Strain (production host): BL21(DE3) / References: UniProt: Q05H60
#2: Chemical ChemComp-EDO / 1,2-ETHANEDIOL / ETHYLENE GLYCOL


Mass: 62.068 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: C2H6O2
#3: Chemical ChemComp-PGE / TRIETHYLENE GLYCOL


Mass: 150.173 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: C6H14O4
#4: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 55 / Source method: isolated from a natural source / Formula: H2O

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Experimental details

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Experiment

ExperimentMethod: X-RAY DIFFRACTION

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Sample preparation

CrystalDensity Matthews: 2.33 Å3/Da / Density % sol: 47.14 %
Crystal growMethod: vapor diffusion / Details: Ammonium sulphate, PEG 400, VAPOR DIFFUSION

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Data collection

Diffraction sourceSource: SYNCHROTRON / Site: ESRF / Beamline: ID23-1
DetectorType: ADSC QUANTUM 315 / Detector: CCD
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthRelative weight: 1
ReflectionResolution: 2.06→59.23 Å / Num. obs: 14086 / Observed criterion σ(I): 1.8 / Redundancy: 5.3 % / Rmerge(I) obs: 0.087 / Net I/σ(I): 7.2
Reflection shellResolution: 2.06→2.206 Å / Redundancy: 5.5 % / Rmerge(I) obs: 0.421 / Mean I/σ(I) obs: 4 / Num. unique all: 2416

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Processing

SoftwareName: REFMAC / Version: 5.2.0019 / Classification: refinement
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: NT wild-type

Resolution: 2.15→59.2 Å / Cor.coef. Fo:Fc: 0.952 / Cor.coef. Fo:Fc free: 0.913 / SU B: 9.362 / SU ML: 0.12 / Cross valid method: THROUGHOUT / ESU R: 0.208 / ESU R Free: 0.186 / Stereochemistry target values: MAXIMUM LIKELIHOOD / Details: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS
RfactorNum. reflection% reflectionSelection details
Rfree0.22841 739 5 %RANDOM
Rwork0.17013 ---
obs0.17299 14086 99.52 %-
Solvent computationIon probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.2 Å / Solvent model: MASK
Displacement parametersBiso mean: 14.606 Å2
Baniso -1Baniso -2Baniso -3
1-0.78 Å20.39 Å20 Å2
2--0.78 Å20 Å2
3----1.17 Å2
Refinement stepCycle: LAST / Resolution: 2.15→59.2 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms1856 0 18 55 1929
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.0160.0221933
X-RAY DIFFRACTIONr_angle_refined_deg1.4231.9432619
X-RAY DIFFRACTIONr_dihedral_angle_1_deg4.8945269
X-RAY DIFFRACTIONr_dihedral_angle_2_deg38.98627.12373
X-RAY DIFFRACTIONr_dihedral_angle_3_deg14.62815320
X-RAY DIFFRACTIONr_dihedral_angle_4_deg10.151152
X-RAY DIFFRACTIONr_chiral_restr0.090.2302
X-RAY DIFFRACTIONr_gen_planes_refined0.0060.021450
X-RAY DIFFRACTIONr_nbd_refined0.2170.2924
X-RAY DIFFRACTIONr_nbtor_refined0.2950.21389
X-RAY DIFFRACTIONr_xyhbond_nbd_refined0.130.267
X-RAY DIFFRACTIONr_symmetry_vdw_refined0.1940.235
X-RAY DIFFRACTIONr_symmetry_hbond_refined0.1620.25
X-RAY DIFFRACTIONr_mcbond_it0.8891.51326
X-RAY DIFFRACTIONr_mcangle_it1.13322074
X-RAY DIFFRACTIONr_scbond_it2.2363674
X-RAY DIFFRACTIONr_scangle_it3.0164.5540
LS refinement shellResolution: 2.15→2.206 Å / Total num. of bins used: 20
RfactorNum. reflection% reflection
Rfree0.238 59 -
Rwork0.161 1037 -
obs--99.73 %
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)
123.8146-5.18152.185724.0284-6.734428.6759-0.421-0.4670.479-0.0467-0.5039-2.00580.25742.04890.92480.0658-0.0253-0.03160.19150.09980.2308-13.21320.729816.9536
29.1728-5.37980.166513.71353.42357.0058-0.14570.18050.04020.1918-0.2935-0.2130.4223-0.35190.43920.0836-0.0763-0.00870.0694-0.0003-0.0009-22.329718.448817.1607
36.9593-6.41435.540915.9052-2.89664.9006-0.1257-0.3067-0.23040.2053-0.0962-0.0712-0.1695-0.66330.22190.0364-0.0407-0.02240.131-0.04340.0455-25.343126.336419.3487
42.7057-8.1332-3.671128.62670.431531.89160.2885-0.6578-0.0293-0.17490.4891.2137-0.2379-0.7195-0.77740.16620.0193-0.03940.1492-0.0270.0938-27.888334.261320.5755
518.37847.12983.94039.5991-2.90393.72-0.1877-1.0144-0.08160.5213-0.1711-0.0699-0.4242-0.36270.35880.21410.1009-0.04410.0408-0.10860.1152-29.988542.832118.3943
65.7829-5.92942.762821.1424.53737.2775-0.0546-0.36090.03240.34210.27460.2878-0.3953-0.2179-0.21990.08970.0630.04020.0517-0.03890.026-33.290335.341512.9133
78.8062-7.82697.002250.8484-5.73555.5731-0.5120.16070.0109-0.06230.21461.2887-0.4436-0.7920.29740.09020.01630.0130.14960.03220.0572-32.473523.817411.7162
87.961-0.04525.873815.22519.694310.5492-0.19630.1175-0.24270.3141-0.04880.0671-0.0815-0.63010.24510.1341-0.04230.08980.07280.03580.0093-29.722215.360811.0047
91.44393.781-1.790813.7808-1.53494.78590.16270.1242-0.2216-0.0119-0.046-0.06370.28130.0626-0.11670.19660.00480.01940.08980.01720.135-23.902312.63727.6623
104.90390.05281.829517.49363.44547.50820.34050.5389-0.0383-0.4481-0.4628-0.3841-0.03160.36190.12230.1031-0.02260.00150.1224-0.01290.0176-19.347920.33957.445
116.618-1.9088-0.62134.96570.49170.0611-0.0442-0.30510.48070.5232-0.3984-0.1521-0.06740.61740.44270.1941-0.0150.01050.1521-0.0060.0375-22.422229.3399.3506
127.6077-4.5319-1.61342.89550.56631.13810.1068-0.13830.01660.2313-0.5831-0.2326-0.2263-0.45410.47630.2278-0.0444-0.02880.1146-0.06780.095-26.456837.60388.3446
136.76026.3074-3.0467.179-3.42941.6391-0.11970.39040.34630.1156-0.0488-0.0356-0.0291-0.38420.16850.3360.0211-0.05890.0843-0.01120.2552-28.954746.67959.4152
142.5686-3.26791.738233.0879-6.81152.2465-0.0092-0.25950.7414-0.5161-0.1628-0.9078-0.64390.12920.17190.2707-0.0686-0.05080.0134-0.03860.1447-21.184245.029915.5482
151.4374-0.6705-1.433611.9162-8.36648.46530.1954-0.26550.257-0.3612-0.5479-0.3431-0.3053-0.08190.35260.1221-0.032-0.07290.0996-0.05890.0781-19.57135.576319.0489
1618.96191.65760.58520.0974-7.9543.22980.0169-0.91380.26861.20810.01790.23370.1457-0.0952-0.03480.0936-0.0858-0.07550.1081-0.03410.0546-19.070729.177924.1559
1714.9308-0.8936-0.54318.1804-6.37545.0724-0.17240.3589-0.5802-0.3394-0.4533-0.50470.53220.25510.62580.0992-0.0049-0.03510.0951-0.0480.0392-14.630826.268222.0079
1812.2771.5924-5.176811.5503-10.40122.8988-0.09310.0092-0.1459-0.1206-0.5383-0.88790.38881.32370.63140.047-0.02580.00240.15980.07620.0869-12.55933.153813.5884
197.0451-6.97232.923.0273-4.115511.2874-0.28530.11340.7096-0.4141-0.348-0.7957-0.43970.71450.63330.0665-0.1353-0.030.05430.04280.0505-17.806539.84116.6731
2013.4228-3.9494-4.00451.1661.378411.3079-0.2635-0.0470.867-0.844-0.26151.03-1.1482-1.24360.5250.4634-0.0382-0.14570.16830.10550.2968-26.039443.2676-3.7251
2120.55914.796212.333314.9951-4.908215.1852-0.32420.0670.30860.15560.1057-0.0228-0.6439-0.0310.21850.06620.009-0.00280.0761-0.02870.0541-35.380836.5399-6.3072
225.5361-3.56651.322210.0551.91781.30460.124-0.0658-0.0195-0.1016-0.24480.308-0.206-0.12910.12080.0637-0.0249-0.01080.07560.0040.0271-35.438929.742-6.095
239.8436-2.5638-3.44999.9184.44158.6586-0.1846-0.00040.15450.16940.06260.29680.4658-0.00620.1220.052-0.04950.01940.05760.0263-0.0282-27.292323.4497-7.6781
2438.5545-15.1998-19.884314.85862.77313.1501-0.32382.5794-2.071-0.36620.01630.5620.5112-0.35560.30750.1443-0.0234-0.00410.118-0.04610.1469-22.43119.9432-12.7216
2513.24046.2425-3.52965.0077-3.07841.90970.1705-0.1023-0.8124-0.0394-0.1278-0.17840.48890.582-0.04270.09360.06640.06040.11060.01970.1412-14.227717.0262-6.81
2613.3673-7.2668-2.674110.8733-0.023817.88570.199-0.6617-0.5721-0.3334-0.0791-0.10221.01760.0122-0.11990.0955-0.03150.01570.03040.08630.046-19.137317.1302-2.3922
2719.9203-6.6732-8.921515.4923-3.13556.8248-0.0119-0.5479-0.26530.0230.02760.1102-0.09360.0959-0.01570.1352-0.0006-0.01060.14310.00780.0585-27.397818.74470.3477
2823.3607-10.15877.02587.52212.740812.9342-0.26060.16880.45830.73520.3663-0.15310.62430.0784-0.10580.1046-0.02310.01360.0690.03160.0629-33.196623.56731.7925
2915.1918-4.6858-2.98526.99475.79147.6724-0.14580.0314-0.24690.6201-0.08260.05140.66340.13070.22850.0854-0.00410.05350.08540.04680.0659-41.459527.17645.3174
3014.15582.565610.11482.0178-1.436514.1126-0.2456-0.09220.52480.36130.3620.3397-0.8066-0.1978-0.11640.12320.03890.0240.0524-0.01290.1232-36.183836.0633.2168
3117.2123-4.4507-3.83866.26474.55684.7689-0.044-0.12230.13790.2284-0.13520.1066-0.21460.24270.17920.0631-0.03070.02230.0814-0.00620.0209-25.828631.06441.0222
3219.8517-1.2644-0.203710.5299-0.56550.0341-0.1457-0.6939-0.47830.2873-0.1974-0.18020.31620.13420.34310.12920.01190.02080.1287-0.01130.0385-15.90624.86580.4006
333.01644.0679-0.168410.02751.87240.9799-0.158-0.53870.00050.54730.0052-0.85350.25590.55220.15270.15340.0635-0.01720.33670.08110.1583-9.18121.0007-2.5218
3420.1933-4.76624.6528.75712.657612.8799-0.03150.35840.3391-0.3978-0.0863-0.3742-0.12680.78790.11780.0246-0.02820.08180.07660.06030.0438-11.749626.1672-9.5952
3514.1150.19473.26655.1488-0.30010.7791-0.28380.00870.4279-0.2545-0.0259-0.25110.2111-0.15950.30970.1203-0.01250.03720.10820.01960.0758-21.441728.6779-11.9425
3618.09682.24336.28490.49922.04779.4610.1272-0.0227-0.3867-0.130.1358-0.1566-0.1024-0.5319-0.2630.0747-0.0435-0.00070.09710.01110.0564-29.749430.5937-13.815
377.39630.56424.26420.16921.16127.99740.1791-0.09790.6045-0.0032-0.07260.1663-0.4503-0.3662-0.10650.0682-0.02610.00590.07620.03380.0935-25.937737.6423-12.7725
3810.89480.35498.48118.7601-5.284610.13680.2273-0.7022-0.01090.1654-0.18770.1278-0.1172-0.4437-0.03950.0336-0.0580.01480.0853-0.01340.058-19.79736.2375-5.9688
399.8312-3.7317.35818.9928-9.452811.36190.0765-0.30330.0420.1955-0.3549-0.1086-0.1787-0.09680.27840.0672-0.09890.03670.1494-0.00790.0555-13.036933.9858-1.4987
4043.9332-16.06229.048618.89048.098211.8580.0184-1.57051.23510.8420.5033-2.0137-0.74621.5858-0.52170.0516-0.1638-0.0710.44910.07030.1688-7.74630.5674.2413
Refinement TLS group
IDRefine-IDRefine TLS-IDAuth asym-IDAuth seq-ID
1X-RAY DIFFRACTION1A5 - 10
2X-RAY DIFFRACTION2A11 - 17
3X-RAY DIFFRACTION3A18 - 22
4X-RAY DIFFRACTION4A23 - 28
5X-RAY DIFFRACTION5A29 - 36
6X-RAY DIFFRACTION6A37 - 45
7X-RAY DIFFRACTION7A46 - 51
8X-RAY DIFFRACTION8A52 - 57
9X-RAY DIFFRACTION9A58 - 63
10X-RAY DIFFRACTION10A64 - 69
11X-RAY DIFFRACTION11A70 - 75
12X-RAY DIFFRACTION12A76 - 81
13X-RAY DIFFRACTION13A82 - 89
14X-RAY DIFFRACTION14A90 - 96
15X-RAY DIFFRACTION15A97 - 102
16X-RAY DIFFRACTION16A103 - 107
17X-RAY DIFFRACTION17A108 - 113
18X-RAY DIFFRACTION18A114 - 119
19X-RAY DIFFRACTION19A120 - 128
20X-RAY DIFFRACTION20A129 - 135
21X-RAY DIFFRACTION21B6 - 10
22X-RAY DIFFRACTION22B11 - 19
23X-RAY DIFFRACTION23B20 - 24
24X-RAY DIFFRACTION24B25 - 30
25X-RAY DIFFRACTION25B31 - 36
26X-RAY DIFFRACTION26B37 - 41
27X-RAY DIFFRACTION27B42 - 47
28X-RAY DIFFRACTION28B48 - 52
29X-RAY DIFFRACTION29B53 - 60
30X-RAY DIFFRACTION30B61 - 66
31X-RAY DIFFRACTION31B67 - 76
32X-RAY DIFFRACTION32B77 - 82
33X-RAY DIFFRACTION33B83 - 90
34X-RAY DIFFRACTION34B91 - 96
35X-RAY DIFFRACTION35B97 - 103
36X-RAY DIFFRACTION36B104 - 108
37X-RAY DIFFRACTION37B109 - 115
38X-RAY DIFFRACTION38B116 - 120
39X-RAY DIFFRACTION39B121 - 126
40X-RAY DIFFRACTION40B127 - 132

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