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- PDB-6fuz: Crystal structure of the TPR domain of KLC1 in complex with the C... -

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Basic information

Entry
Database: PDB / ID: 6fuz
TitleCrystal structure of the TPR domain of KLC1 in complex with the C-terminal peptide of JIP1
Components
  • Kinesin light chain 1,Kinesin light chain 1,C-Jun-amino-terminal kinase-interacting protein 1
  • nanobody
KeywordsMOTOR PROTEIN / Protein complex / nanobody / cargo recognition
Function / homology
Function and homology information


RHO GTPases activate KTN1 / membrane-bounded organelle / Kinesins / COPI-dependent Golgi-to-ER retrograde traffic / protein localization to synapse / MHC class II antigen presentation / axo-dendritic transport / stress granule disassembly / ciliary rootlet / kinesin complex ...RHO GTPases activate KTN1 / membrane-bounded organelle / Kinesins / COPI-dependent Golgi-to-ER retrograde traffic / protein localization to synapse / MHC class II antigen presentation / axo-dendritic transport / stress granule disassembly / ciliary rootlet / kinesin complex / microtubule-based movement / kinesin binding / tubulin binding / intracellular protein transport / growth cone / cytoplasmic vesicle / vesicle / microtubule / cell adhesion / neuron projection / axon / neuronal cell body / membrane / cytosol / cytoplasm
Similarity search - Function
Kinesin light chain / Kinesin light chain repeat / Kinesin light chain repeat. / Tetratricopeptide repeat / Tetratricopeptide repeat / Tetratricopeptide repeat / Tetratricopeptide repeat domain / TPR repeat region circular profile. / TPR repeat profile. / Tetratricopeptide repeats ...Kinesin light chain / Kinesin light chain repeat / Kinesin light chain repeat. / Tetratricopeptide repeat / Tetratricopeptide repeat / Tetratricopeptide repeat / Tetratricopeptide repeat domain / TPR repeat region circular profile. / TPR repeat profile. / Tetratricopeptide repeats / Tetratricopeptide repeat / Serine Threonine Protein Phosphatase 5, Tetratricopeptide repeat / Alpha Horseshoe / Tetratricopeptide-like helical domain superfamily / Mainly Alpha
Similarity search - Domain/homology
Kinesin light chain 1 / Kinesin light chain
Similarity search - Component
Biological speciesMus musculus (house mouse)
Homo sapiens (human)
Lama glama (llama)
MethodX-RAY DIFFRACTION / SYNCHROTRON / Resolution: 2.7 Å
AuthorsPernigo, S. / Dodding, M.P. / Steiner, R.A.
Funding support United Kingdom, 1items
OrganizationGrant numberCountry
Biotechnology and Biological Sciences Research CouncilBB/L006774/1 United Kingdom
Citation
Journal: Elife / Year: 2018
Title: Structural basis for isoform-specific kinesin-1 recognition of Y-acidic cargo adaptors.
Authors: Pernigo, S. / Chegkazi, M.S. / Yip, Y.Y. / Treacy, C. / Glorani, G. / Hansen, K. / Politis, A. / Bui, S. / Dodding, M.P. / Steiner, R.A.
#1: Journal: Science / Year: 2013
Title: Structural basis for kinesin-1:cargo recognition.
Authors: Pernigo, S. / Lamprecht, A. / Steiner, R.A. / Dodding, M.P.
History
DepositionFeb 28, 2018Deposition site: PDBE / Processing site: PDBE
Revision 1.0May 2, 2018Provider: repository / Type: Initial release
Revision 1.1Sep 12, 2018Group: Data collection / Database references / Source and taxonomy
Category: citation / entity_src_gen
Item: _citation.title / _entity_src_gen.pdbx_gene_src_ncbi_taxonomy_id / _entity_src_gen.pdbx_gene_src_scientific_name
Revision 1.2Sep 18, 2019Group: Data collection / Database references / Category: citation / citation_author
Item: _citation.country / _citation.journal_abbrev ..._citation.country / _citation.journal_abbrev / _citation.journal_id_CSD / _citation.journal_id_ISSN / _citation.journal_volume / _citation.pdbx_database_id_DOI / _citation.pdbx_database_id_PubMed / _citation.title / _citation.year
Revision 1.3Oct 16, 2024Group: Data collection / Database references / Structure summary
Category: chem_comp_atom / chem_comp_bond ...chem_comp_atom / chem_comp_bond / database_2 / pdbx_entry_details / pdbx_modification_feature
Item: _database_2.pdbx_DOI / _database_2.pdbx_database_accession

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

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Assembly

Deposited unit
A: Kinesin light chain 1,Kinesin light chain 1,C-Jun-amino-terminal kinase-interacting protein 1
N: nanobody
hetero molecules


Theoretical massNumber of molelcules
Total (without water)50,7903
Polymers50,6982
Non-polymers921
Water23413
1


  • Idetical with deposited unit
  • defined by author&software
  • Evidence: gel filtration, isothermal titration calorimetry, assay for oligomerization, Fluorescence polarisation
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area1850 Å2
ΔGint-7 kcal/mol
Surface area17610 Å2
MethodPISA
Unit cell
Length a, b, c (Å)107.473, 90.364, 51.698
Angle α, β, γ (deg.)90.00, 99.80, 90.00
Int Tables number5
Space group name H-MC121

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Components

#1: Protein Kinesin light chain 1,Kinesin light chain 1,C-Jun-amino-terminal kinase-interacting protein 1 / JNK-interacting protein 1 / Islet-brain 1 / IB-1 / JNK MAP kinase scaffold protein 1 / Mitogen- ...JNK-interacting protein 1 / Islet-brain 1 / IB-1 / JNK MAP kinase scaffold protein 1 / Mitogen-activated protein kinase 8-interacting protein 1


Mass: 37301.824 Da / Num. of mol.: 1
Source method: isolated from a genetically manipulated source
Details: Chains A is a chimeric construct of TPR domain of KLC1 and J the C-terminal peptide of JIP1 fused via a (TGS)10 linker.,Chains A is a chimeric construct of TPR domain of KLC1 and J the C- ...Details: Chains A is a chimeric construct of TPR domain of KLC1 and J the C-terminal peptide of JIP1 fused via a (TGS)10 linker.,Chains A is a chimeric construct of TPR domain of KLC1 and J the C-terminal peptide of JIP1 fused via a (TGS)10 linker.
Source: (gene. exp.) Mus musculus (house mouse), (gene. exp.) Homo sapiens (human)
Gene: Klc1, Kns2, MAPK8IP1, IB1, JIP1, PRKM8IP / Production host: Escherichia coli BL21 (bacteria) / References: UniProt: Q5UE59, UniProt: O88447*PLUS
#2: Antibody nanobody


Mass: 13395.817 Da / Num. of mol.: 1
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Lama glama (llama) / Production host: Escherichia coli (E. coli) / Variant (production host): WK6
#3: Chemical ChemComp-GOL / GLYCEROL / GLYCERIN / PROPANE-1,2,3-TRIOL


Mass: 92.094 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: C3H8O3
#4: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 13 / Source method: isolated from a natural source / Formula: H2O
Has protein modificationY

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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: 2.44 Å3/Da / Density % sol: 49.58 %
Crystal growTemperature: 291.15 K / Method: vapor diffusion, hanging drop
Details: 0.04 M potassium phosphate monobasic, 16% PEG8000, 20% glycerol

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: PETRA III, EMBL c/o DESY / Beamline: P14 (MX2) / Wavelength: 0.9763 Å
DetectorType: DECTRIS PILATUS 6M-F / Detector: PIXEL / Date: Oct 27, 2016
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.9763 Å / Relative weight: 1
ReflectionResolution: 2.7→45.18 Å / Num. obs: 13229 / % possible obs: 98.6 % / Redundancy: 3.8 % / Biso Wilson estimate: 94.37 Å2 / CC1/2: 0.999 / Rmerge(I) obs: 0.051 / Rpim(I) all: 0.047 / Net I/σ(I): 11.9
Reflection shellResolution: 2.7→2.83 Å / Rmerge(I) obs: 1.176 / Mean I/σ(I) obs: 1 / Num. unique obs: 1747 / CC1/2: 0.584 / Rpim(I) all: 1.067 / % possible all: 98.5

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Processing

Software
NameVersionClassification
BUSTER2.10.3refinement
XDSdata reduction
XSCALEdata scaling
PHASERphasing
RefinementResolution: 2.7→45.18 Å / Cor.coef. Fo:Fc: 0.926 / Cor.coef. Fo:Fc free: 0.933 / Rfactor Rfree error: 0 / SU R Cruickshank DPI: 1.024 / Cross valid method: THROUGHOUT / σ(F): 0 / SU R Blow DPI: 0.822 / SU Rfree Blow DPI: 0.32 / SU Rfree Cruickshank DPI: 0.333
RfactorNum. reflection% reflectionSelection details
Rfree0.257 669 5.06 %RANDOM
Rwork0.233 ---
obs0.234 13228 98.4 %-
Displacement parametersBiso mean: 113.06 Å2
Baniso -1Baniso -2Baniso -3
1-11.0292 Å20 Å2-23.036 Å2
2---3.6355 Å20 Å2
3----7.3937 Å2
Refine analyzeLuzzati coordinate error obs: 0.48 Å
Refinement stepCycle: 1 / Resolution: 2.7→45.18 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms3001 0 6 13 3020
Refine LS restraints
Refine-IDTypeDev idealNumberRestraint functionWeight
X-RAY DIFFRACTIONt_bond_d0.0083060HARMONIC2
X-RAY DIFFRACTIONt_angle_deg1.074129HARMONIC2
X-RAY DIFFRACTIONt_dihedral_angle_d1091SINUSOIDAL2
X-RAY DIFFRACTIONt_incorr_chiral_ct
X-RAY DIFFRACTIONt_pseud_angle
X-RAY DIFFRACTIONt_trig_c_planes88HARMONIC2
X-RAY DIFFRACTIONt_gen_planes436HARMONIC5
X-RAY DIFFRACTIONt_it3060HARMONIC20
X-RAY DIFFRACTIONt_nbd
X-RAY DIFFRACTIONt_omega_torsion2.45
X-RAY DIFFRACTIONt_other_torsion19.92
X-RAY DIFFRACTIONt_improper_torsion
X-RAY DIFFRACTIONt_chiral_improper_torsion386SEMIHARMONIC5
X-RAY DIFFRACTIONt_sum_occupancies
X-RAY DIFFRACTIONt_utility_distance
X-RAY DIFFRACTIONt_utility_angle
X-RAY DIFFRACTIONt_utility_torsion
X-RAY DIFFRACTIONt_ideal_dist_contact3493SEMIHARMONIC4
LS refinement shellResolution: 2.7→2.92 Å / Rfactor Rfree error: 0 / Total num. of bins used: 7
RfactorNum. reflection% reflection
Rfree0.238 152 5.64 %
Rwork0.238 2545 -
all0.238 2697 -
obs--98 %
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)
1-0.36092.98761.17720.05131.31790.43250.0046-0.0206-0.10580.01050.0387-0.01850.0917-0.1318-0.0433-0.4341-0.313-0.21260.43960.0262-0.559347.58621.56792.8758
2-0.0499-2.49033.07470.4753.6382.5648-0.0536-0.15480.1960.20740.0709-0.0562-0.0586-0.104-0.0173-0.2867-0.23290.23230.6035-0.34870.193447.392412.228-0.7543
37.18024.6766-6.36649.8144-5.05728.5654-0.06650.0837-0.1339-0.1617-0.0045-0.40980.33440.82610.071-0.18370.0166-0.01120.31470.0333-0.193144.1458-0.2553-8.9499
411.3382-4.5442-2.92524.6827-6.62965.3141-0.09610.82690.3421-0.015-0.00450.2288-0.1320.02410.1006-0.2281-0.02290.0880.09640.1381-0.132436.58184.1298-11.6006
58.5574-1.2119-2.13870.32032.31346.7221-0.19330.8585-0.0611-0.49440.14050.03010.50060.09950.0528-0.0913-0.10020.0405-0.07590.0863-0.20231.7752-3.8179-7.6994
6-1.58860.38014.01862.14975.06376.71350.10390.6901-0.2176-0.08580.1087-0.0203-0.1526-0.2233-0.2126-0.1778-0.10070.07870.39930.0251-0.087422.01821.99-10.4728
70.73952.7315-1.87940.0025-0.2302-0.67130.0182-0.0021-0.1961-0.01190.04290.0980.17420.0451-0.061-0.14630.03430.2307-0.28080.1147-0.015925.2209-8.8966-2.8823
81.2305-1.0987-3.51553.0183-0.177-0.53660.0248-0.0066-0.37950.00090.01390.03610.3897-0.0499-0.03870.04280.12190.2729-0.02370.30290.31225.5061-9.1183.7772
91.7892-0.78910.97380.03010.82313.4337-0.10580.5636-0.00370.1488-0.17210.79540.2998-0.27790.278-0.2601-0.04690.0459-0.03330.21320.082614.26392.3144-1.3536
100.6908-4.5301-2.15270.21541.88131.7718-0.0932-0.5025-0.11260.7209-0.15160.0510.19430.01980.24480.05860.00740.3095-0.06480.2668-0.165820.3104-3.015510.8224
111.28880.7482-3.357702.0979-0.47660.0008-0.0810.16860.3594-0.02090.0109-0.00750.18140.0201-0.1886-0.1075-0.0163-0.22070.2258-0.104922.21615.452711.3422
121.3688-1.4579-4.1176.9711-0.02721.7909-0.02630.40280.21870.1190.06940.4738-0.3057-0.2217-0.0431-0.2013-0.01530.088-0.25790.25080.139311.283811.333410.9234
130.53361.6551.49460.1913-4.0141.48630.0059-0.13660.0387-0.0285-0.05250.02720.01190.0710.04650.1533-0.2277-0.26520.43060.254-0.144824.134310.764425.5831
14-0.7879-0.29032.20950-2.93040.825-0.0549-0.12910.17330.13610.1473-0.1546-0.01890.1514-0.09240.1358-0.2312-0.0119-0.27820.0987-0.049316.302116.413528.6715
15-0.08250.60570.15140.69660.91161.0121-0.0148-0.08470.02220.04060.02770.0405-0.0098-0.0448-0.01290.1546-0.13490.1214-0.09020.1364-0.12179.757525.098724.5893
161.8772.74060.06233.3385-0.5203-0.0340.01050.07320.11530.06350.2095-0.0041-0.13550.3248-0.2201-0.10910.0413-0.035-0.15580.2782-0.197920.333218.008313.3619
170.6908-0.0072-1.10112.3677-0.45470.5053-0.01290.0778-0.0008-0.01790.01280.1345-0.03540.0050.0001-0.07680.04220.2387-0.07320.27750.237910.040423.548111.5781
182.09521.17940.6661.6879-4.91581.76310.0066-0.00410.22460.2020.0447-0.0836-0.0398-0.1392-0.05140.16340.12460.1226-0.19890.2791-0.113116.001626.743817.0313
190.23760.0251-0.682500.3347-0.05980.0071-0.0252-0.00640.02010.0371-0.0034-0.0203-0.052-0.04410.044-0.167-0.088-0.0143-0.18640.084829.062424.081715.8698
20-0.5694-3.49352.16222.0607-6.07090.76390.0065-0.03390.02340.1519-0.045-0.0525-0.04710.11650.0385-0.11990.19190.067-0.04380.33090.129414.735-13.594226.8034
210.0622-2.67842.132802.6090.9346-0.02090.0318-0.0305-0.04420.0049-0.17740.0581-0.13130.0160.0904-0.09770.2734-0.49410.0620.3458-5.6333-20.531121.4725
221.6481-0.3891-1.994800.6692-0.7383-0.00930.0038-0.0018-0.043-0.04590.13730.1907-0.07570.0552-0.34980.01220.2066-0.40280.07140.3021-2.4133-12.997223.091
230.4764-0.8282-0.64050.0445-0.6141-0.20140.01990.0211-0.0386-0.0602-0.0504-0.1218-0.14830.2710.0305-0.02330.1353-0.03980.22240.1925-0.01215.3946-8.409627.8598
240.4035-0.0985-1.15560.1898-1.1501-0.1279-0.03-0.0190.1285-0.0520.0389-0.10440.01150.1753-0.00890.1109-0.09470.1590.32510.286-0.130517.6608-1.04224.0066
253.32123.2481.33550.11621.16730.3928-0.09350.0641-0.3488-0.1367-0.17010.14630.23550.76350.2636-0.34980.05780.2282-0.42850.03920.180910.3061-10.538315.533
261.1937-1.3674-2.89670.42572.9532-0.86230.04110.1169-0.19-0.0771-0.02530.05170.0832-0.0603-0.01580.13370.18250.2451-0.4944-0.05330.54648.4906-21.99129.1642
271.092-2.21743.002301.78141.01210.02130.0655-0.15030.0749-0.1086-0.0707-0.12730.17580.0874-0.4759-0.05250.20220.08670.29110.09218.6241-5.200414.1535
280.98440.6562-2.76453.07813.1701-0.9844-0.00190.02940.0470.2758-0.0398-0.1537-0.2823-0.04260.0417-0.1675-0.08250.2213-0.1210.06210.36628.02892.074515.8109
291.8825-3.16440.79170.98482.27710.21730.01230.01620.0153-0.22530.03070.01250.03020.0173-0.043-0.1031-0.0521-0.06040.1938-0.01490.02931.4742-7.96869.3929
30-0.50251.38391.54262.5274-2.67541.54750.03570.15050.0418-0.0101-0.0286-0.0775-0.2433-0.0148-0.0071-0.17960.0512-0.02550.0646-0.0960.1238-1.3178-6.945314.6155
31-0.18911.99360.532103.42480.18940.036-0.1798-0.02990.1594-0.03750.0015-0.0929-0.06470.0015-0.011-0.12960.17060.09990.02720.15878.9205-1.512728.3411
32-0.037-2.45180.13863.37393.80430.54660.0047-0.10340.06770.1390.0987-0.09770.0362-0.014-0.1034-0.24090.05130.125-0.09270.2093-0.09717.4274-6.443525.8862
330.6307-2.69863.54190.00571.55661.20480.01690.3181-0.2616-0.1841-0.09460.02110.2174-0.06570.0777-0.2619-0.06610.1067-0.2105-0.23590.3791-4.696-13.468116.5802
34-0.5429-2.23411.508900.57440.7267-0.09260.05770.0033-0.04320.1030.18580.20060.0097-0.0104-0.09710.03450.225-0.49920.03130.42332.6529-18.513614.8916
352.0729-0.1077-1.1860.7935-3.2806-0.4705-0.03510.0055-0.14170.2447-0.1026-0.16610.02110.02660.1377-0.15290.04890.26720.28760.12180.127114.9404-11.861416.0566
360.872-2.6679-2.44891.68490.4046-0.872-0.02470.15720.06160.0012-0.04280.0262-0.08580.21430.0675-0.26160.27390.20020.08750.15160.325817.9088-15.754720.2058
370.5965-1.383-1.12231.7865-0.95521.0202-0.0057-0.089-0.0816-0.13850.07060.09510.1340.1896-0.0649-0.02960.01870.2337-0.50540.09640.46381.9255-21.412320.4
387.81415.96764.17012.1278-0.35846.98810.0283-0.09530.32020.18890.05240.0757-0.25780.2204-0.0807-0.0689-0.06540.0431-0.10310.0760.038623.84259.98972.3064
Refinement TLS group
IDRefine-IDRefine TLS-IDSelection details
1X-RAY DIFFRACTION1{ A|214 - A|222 }
2X-RAY DIFFRACTION2{ A|223 - A|236 }
3X-RAY DIFFRACTION3{ A|245 - A|266 }
4X-RAY DIFFRACTION4{ A|267 - A|286 }
5X-RAY DIFFRACTION5{ A|287 - A|309 }
6X-RAY DIFFRACTION6{ A|310 - A|322 }
7X-RAY DIFFRACTION7{ A|323 - A|327 }
8X-RAY DIFFRACTION8{ A|328 - A|341 }
9X-RAY DIFFRACTION9{ A|342 - A|360 }
10X-RAY DIFFRACTION10{ A|361 - A|375 }
11X-RAY DIFFRACTION11{ A|376 - A|387 }
12X-RAY DIFFRACTION12{ A|388 - A|409 }
13X-RAY DIFFRACTION13{ A|410 - A|417 }
14X-RAY DIFFRACTION14{ A|418 - A|430 }
15X-RAY DIFFRACTION15{ A|431 - A|435 }
16X-RAY DIFFRACTION16{ A|436 - A|473 }
17X-RAY DIFFRACTION17{ A|474 - A|478 }
18X-RAY DIFFRACTION18{ A|479 - A|490 }
19X-RAY DIFFRACTION19{ A|491 - A|495 }
20X-RAY DIFFRACTION20{ N|2 - N|7 }
21X-RAY DIFFRACTION21{ N|8 - N|14 }
22X-RAY DIFFRACTION22{ N|15 - N|20 }
23X-RAY DIFFRACTION23{ N|21 - N|27 }
24X-RAY DIFFRACTION24{ N|28 - N|32 }
25X-RAY DIFFRACTION25{ N|33 - N|40 }
26X-RAY DIFFRACTION26{ N|41 - N|46 }
27X-RAY DIFFRACTION27{ N|47 - N|52 }
28X-RAY DIFFRACTION28{ N|53 - N|59 }
29X-RAY DIFFRACTION29{ N|60 - N|64 }
30X-RAY DIFFRACTION30{ N|65 - N|70 }
31X-RAY DIFFRACTION31{ N|71 - N|76 }
32X-RAY DIFFRACTION32{ N|77 - N|81 }
33X-RAY DIFFRACTION33{ N|82 - N|88 }
34X-RAY DIFFRACTION34{ N|89 - N|94 }
35X-RAY DIFFRACTION35{ N|95 - N|102 }
36X-RAY DIFFRACTION36{ N|103 - N|107 }
37X-RAY DIFFRACTION37{ N|108 - N|113 }
38X-RAY DIFFRACTION38{ A|704 - A|711 }

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