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- PDB-8ifj: Crystal structure of pyrrolysyl-tRNA synthetase from methanogenic... -

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Basic information

Entry
Database: PDB / ID: 8ifj
TitleCrystal structure of pyrrolysyl-tRNA synthetase from methanogenic archaeon ISO4-G1
ComponentsPyrrolysyl-tRNA synthetase PylS
KeywordsTRANSLATION / non-canonical amino acids / genetic code expansion / tRNA
Function / homology
Function and homology information


tRNA aminoacylation / aminoacyl-tRNA ligase activity / tRNA binding / translation / ATP binding
Similarity search - Function
Pyrrolysyl-tRNA ligase, C-terminal / Phenylalanyl-tRNA synthetase / tRNA synthetases class II core domain (F) / Aminoacyl-tRNA synthetase, class II / Aminoacyl-transfer RNA synthetases class-II family profile.
Similarity search - Domain/homology
Pyrrolysyl-tRNA synthetase PylS
Similarity search - Component
Biological speciesmethanogenic archaeon mixed culture ISO4-G1 (environmental samples)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 2.78 Å
AuthorsYanagisawa, T. / Tanabe, H. / Yokoyama, S.
Funding support Japan, 4items
OrganizationGrant numberCountry
Japan Agency for Medical Research and Development (AMED)JP17am0101081 Japan
Japan Agency for Medical Research and Development (AMED)JP19gm0010001 Japan
Japan Society for the Promotion of Science (JSPS)16K05859 Japan
Japan Society for the Promotion of Science (JSPS)24550203 Japan
CitationJournal: Int J Mol Sci / Year: 2023
Title: Crystal Structure of Pyrrolysyl-tRNA Synthetase from a Methanogenic Archaeon ISO4-G1 and Its Structure-Based Engineering for Highly-Productive Cell-Free Genetic Code Expansion with Non-Canonical Amino Acids.
Authors: Yanagisawa, T. / Seki, E. / Tanabe, H. / Fujii, Y. / Sakamoto, K. / Yokoyama, S.
History
DepositionFeb 18, 2023Deposition site: PDBJ / Processing site: PDBJ
Revision 1.0Mar 15, 2023Provider: repository / Type: Initial release
Revision 1.1Apr 26, 2023Group: 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 1.2May 29, 2024Group: Data collection / Category: chem_comp_atom / chem_comp_bond

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: Pyrrolysyl-tRNA synthetase PylS
B: Pyrrolysyl-tRNA synthetase PylS
C: Pyrrolysyl-tRNA synthetase PylS
D: Pyrrolysyl-tRNA synthetase PylS
E: Pyrrolysyl-tRNA synthetase PylS
F: Pyrrolysyl-tRNA synthetase PylS
G: Pyrrolysyl-tRNA synthetase PylS
H: Pyrrolysyl-tRNA synthetase PylS
I: Pyrrolysyl-tRNA synthetase PylS
J: Pyrrolysyl-tRNA synthetase PylS


Theoretical massNumber of molelcules
Total (without water)312,56510
Polymers312,56510
Non-polymers00
Water88349
1
A: Pyrrolysyl-tRNA synthetase PylS
B: Pyrrolysyl-tRNA synthetase PylS


Theoretical massNumber of molelcules
Total (without water)62,5132
Polymers62,5132
Non-polymers00
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area4210 Å2
ΔGint-25 kcal/mol
Surface area25290 Å2
MethodPISA
2
C: Pyrrolysyl-tRNA synthetase PylS
G: Pyrrolysyl-tRNA synthetase PylS


Theoretical massNumber of molelcules
Total (without water)62,5132
Polymers62,5132
Non-polymers00
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area4240 Å2
ΔGint-22 kcal/mol
Surface area25150 Å2
MethodPISA
3
D: Pyrrolysyl-tRNA synthetase PylS
F: Pyrrolysyl-tRNA synthetase PylS


Theoretical massNumber of molelcules
Total (without water)62,5132
Polymers62,5132
Non-polymers00
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area4210 Å2
ΔGint-26 kcal/mol
Surface area25750 Å2
MethodPISA
4
E: Pyrrolysyl-tRNA synthetase PylS
H: Pyrrolysyl-tRNA synthetase PylS


Theoretical massNumber of molelcules
Total (without water)62,5132
Polymers62,5132
Non-polymers00
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area4280 Å2
ΔGint-23 kcal/mol
Surface area25410 Å2
MethodPISA
5
I: Pyrrolysyl-tRNA synthetase PylS
J: Pyrrolysyl-tRNA synthetase PylS


Theoretical massNumber of molelcules
Total (without water)62,5132
Polymers62,5132
Non-polymers00
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area4320 Å2
ΔGint-22 kcal/mol
Surface area25330 Å2
MethodPISA
Unit cell
Length a, b, c (Å)98.510, 102.680, 349.860
Angle α, β, γ (deg.)90.000, 90.000, 90.000
Int Tables number19
Space group name H-MP212121

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Components

#1: Protein
Pyrrolysyl-tRNA synthetase PylS


Mass: 31256.469 Da / Num. of mol.: 10
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) methanogenic archaeon mixed culture ISO4-G1 (environmental samples)
Gene: AUP07_0651 / Production host: Escherichia coli BL21(DE3) (bacteria) / References: UniProt: A0A126QV54
#2: Water ChemComp-HOH / water / Water


Mass: 18.015 Da / Num. of mol.: 49 / 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: 2.83 Å3/Da / Density % sol: 56.54 %
Crystal growTemperature: 293 K / Method: vapor diffusion / pH: 7.2 / Details: HEPES, PEG 3350, KCl

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Data collection

DiffractionMean temperature: 100 K / Serial crystal experiment: N
Diffraction sourceSource: SYNCHROTRON / Site: SPring-8 / Beamline: BL32XU / Wavelength: 1 Å
DetectorType: DECTRIS EIGER X 9M / Detector: PIXEL / Date: Jun 28, 2019
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 1 Å / Relative weight: 1
ReflectionResolution: 2.78→49.867 Å / Num. obs: 172270 / % possible obs: 99.9 % / Redundancy: 5.988 % / Biso Wilson estimate: 78.526 Å2 / CC1/2: 0.998 / Rmerge(I) obs: 0.145 / Rrim(I) all: 0.159 / Χ2: 1.125 / Net I/σ(I): 9.77 / Num. measured all: 1031551 / Scaling rejects: 7
Reflection shell

Diffraction-ID: 1

Resolution (Å)Redundancy (%)Rmerge(I) obsMean I/σ(I) obsNum. measured obsNum. possibleNum. unique obsCC1/2Rrim(I) all% possible all
2.78-2.856.0661.8661.017738912770127570.6922.04299.9
2.85-2.936.0941.5151.277559712425124050.7681.65799.8
2.93-3.025.9941.1211.747231712076120640.8631.22999.9
3.02-3.115.9140.9252.126952211769117550.8951.01599.9
3.11-3.215.7860.6842.886598811413114050.9350.75299.9
3.21-3.325.5790.5123.796121110982109710.9540.56499.9
3.32-3.455.1030.4174.615415010629106110.950.46499.8
3.45-3.595.2190.3036.235305210197101660.9820.33799.7
3.59-3.755.7730.2598.1756821985198420.9860.28599.9
3.75-3.935.4850.20510.1751397938093700.9920.22699.9
3.93-4.145.4960.15212.7448950890789060.9950.167100
4.14-4.46.3660.13815.9453510840984050.9970.15100
4.4-4.76.4740.12118.751226791379130.9970.131100
4.7-5.086.4310.121.0847612740474040.9970.109100
5.08-5.566.6460.11219.6344912676067580.9970.122100
5.56-6.226.6930.10621.4941077613761370.9970.115100
6.22-7.186.9210.10223.5337465541554130.9970.11100
7.18-8.797.1480.08527.8332660456945690.9970.092100
8.79-12.436.8810.05532.5124214351935190.9980.06100
12.43-49.8676.5690.05232.5712481193519000.9980.05698.2

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Processing

Software
NameVersionClassification
XSCALEdata scaling
PHENIX1.16_3549refinement
PDB_EXTRACT3.27data extraction
XDSdata reduction
MOLREPphasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT / Resolution: 2.78→49.867 Å / SU ML: 0.48 / Cross valid method: THROUGHOUT / σ(F): 1.34 / Phase error: 34.72 / Stereochemistry target values: ML
RfactorNum. reflection% reflection
Rfree0.2962 3798 2.21 %
Rwork0.2326 168163 -
obs0.234 171961 99.72 %
Solvent computationShrinkage radii: 0.9 Å / VDW probe radii: 1.11 Å / Solvent model: FLAT BULK SOLVENT MODEL
Displacement parametersBiso max: 223.18 Å2 / Biso mean: 100.0042 Å2 / Biso min: 27.71 Å2
Refinement stepCycle: final / Resolution: 2.78→49.867 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms21566 0 0 49 21615
Biso mean---57.18 -
Num. residues----2712
LS refinement shell

Refine-ID: X-RAY DIFFRACTION / Rfactor Rfree error: 0

Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection Rwork% reflection obs (%)
2.78-2.81520.41031320.4056224100
2.8152-2.85220.4421330.39866247100
2.8522-2.89130.52841580.39036183100
2.8913-2.93260.33351480.37636214100
2.9326-2.97630.4011240.3424623399
2.9763-3.02280.35651470.31456229100
3.0228-3.07240.31781410.31556213100
3.0724-3.12540.36761400.30446199100
3.1254-3.18220.39061180.30486242100
3.1822-3.24340.351520.30766237100
3.2434-3.30960.42841460.30346258100
3.3096-3.38150.35281530.31056162100
3.3815-3.46020.29231260.28516232100
3.4602-3.54670.36331360.26876243100
3.5467-3.64250.36591550.25576249100
3.6425-3.74970.3011310.24686219100
3.7497-3.87070.26951360.23866185100
3.8707-4.0090.25961470.22396232100
4.009-4.16940.28211400.21736257100
4.1694-4.3590.26521310.20816261100
4.359-4.58870.24721410.19356219100
4.5887-4.8760.21721390.18116286100
4.876-5.25210.24251480.17856210100
5.2521-5.77990.27211460.21526234100
5.7799-6.61460.29711380.21326261100
6.6146-8.32740.28241500.21596237100
8.3274-49.8670.28091420.1764619799
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)
10.88010.4851-1.78971.1623-2.28538.4127-0.31270.3712-0.0192-0.35930.19540.15521.0903-0.93040.12810.5136-0.0577-0.02360.75570.09080.624438.29877.7712-60.7771
28.2725-0.4207-5.14362.6439-0.96846.8967-0.1969-0.2074-0.57190.189-0.06-0.05410.85770.16360.21710.4422-0.0063-0.13640.30480.00750.49546.4049-2.1635-29.5995
32.32561.7475-0.80954.3311-3.49217.8712-0.06560.1433-0.2144-0.50750.49210.31951.5673-1.2976-0.42840.6447-0.1456-0.08610.63720.06220.549434.1217-5.1617-37.4941
41.37880.36121.15330.4496-0.5816.25540.08480.40250.66110.7487-0.3893-0.7132-1.409-0.20480.15021.57820.0352-0.25980.69390.0030.911549.678217.1163-0.0891
51.0775-0.04140.4892.0016-1.45346.5657-0.10460.10260.12250.39620.1224-0.1487-0.6047-0.27950.01990.36020.0332-0.0520.2750.00530.520246.820915.9521-30.3237
62.1715-0.2718-1.68543.7236-1.97477.071-0.1024-0.25210.31190.25740.1156-0.2628-0.55470.11540.16760.43380.00350.03980.337-0.060.624249.721519.1232-33.8843
72.1441-0.1601-0.40294.4674-1.30016.9585-0.0559-0.38611.04990.60360.18410.0007-1.9951-0.0203-0.15331.12120.104-0.02530.6221-0.04450.8744.118933.4474-33.9968
85.2688-0.3384-4.01134.3685-1.51363.74210.0409-1.35390.65390.56520.394-0.1934-1.92931.6595-0.07960.9737-0.1487-0.03330.6748-0.00270.830152.672732.6661-37.3138
90.9051-0.0207-0.15874.8093-2.07067.0469-0.01630.06530.42310.67710.2628-0.0624-0.8878-0.2225-0.09690.40190.06540.01320.40050.02730.577145.213419.5319-27.0492
102.06330.3291-0.25361.6145-0.71744.85250.37280.1211-0.11840.0229-0.2569-0.12240.32470.7728-0.09080.61520.0249-0.03540.69530.00750.55322.996953.7286-26.4983
113.04082.1441-1.02994.6763-2.40256.4750.36920.30480.28930.2603-0.3361-0.4425-0.34161.203-0.05520.5947-0.06350.030.82790.03530.58548.100362.4519-31.7939
122.9137-1.0144-1.27873.26460.82578.96050.00310.2377-0.6082-0.65030.34450.01541.213-0.0174-0.09581.5440.3573-0.27541.0092-0.06890.845834.241349.5372-36.4355
131.944-0.8949-0.26861.461-1.25736.31140.09020.129-0.2804-0.27040.02570.20010.7697-0.1013-0.15280.88290.0086-0.13710.27670.0790.770135.977652.1513-9.3058
141.2755-0.1198-0.50061.4141-0.98145.3667-0.0275-0.6955-0.53220.28910.05550.54651.0443-0.05990.22970.8830.0235-0.12180.47580.15880.962737.958645.11019.7434
154.24442.3306-0.36725.9671-1.92124.6524-0.0521.3508-1.0703-0.26780.33010.00221.316-1.00350.06281.2781-0.2259-0.28770.5061-0.04450.948432.287540.9165-1.504
162.54930.2512-1.67313.52850.44711.81090.36340.1866-0.7975-1.70190.03730.08942.10740.57170.03372.0825-0.2752-0.2311.1765-0.17631.381229.670931.5493-6.2965
170.83391.46530.64062.60110.54387.24680.7482-0.173-1.71160.33680.7605-0.16182.7120.89910.51252.28170.24-0.14340.73220.20891.192940.469232.468111.2484
182.39841.2522-1.29733.269-1.21333.53340.08760.7491-0.6792-0.89570.55990.30931.7715-0.2548-0.23661.4092-0.0102-0.44750.55250.07850.923732.072450.9874-13.1264
192.3654-0.1006-0.15131.1301-0.33244.74370.07540.2712-0.13410.3120.0971-0.05110.29310.9245-0.20150.60210.0629-0.06710.90860.01550.66412.56093.0848-54.8892
203.42351.932-4.27367.5772-3.54026.9265-0.19790.36510.2183-0.12530.3153-0.54640.60031.32380.14930.42930.0241-0.05761.5439-0.07310.64987.21634.5191-74.9786
213.34060.3031-0.85331.22350.30293.1418-0.0790.50170.3430.10380.2894-0.3468-0.14161.8103-0.20060.5199-0.0755-0.02251.5522-0.00170.6110.391212.5926-66.7589
221.4837-0.05780.02422.0465-1.4947.0299-0.014-0.20050.04890.58250.03270.059-0.55880.2535-0.05780.9433-0.00250.02160.33610.0680.726440.97662.730613.6939
230.8295-0.92140.36484.7499-3.08454.3947-0.0199-0.0460.20920.95020.11240.1213-1.19010.0106-0.05021.20360.09170.04030.55860.06730.658535.650373.03971.0026
245.30560.4731-2.94259.94026.13916.4152-0.47770.1439-0.71330.42460.27520.04510.6917-1.05920.21251.1801-0.0482-0.21121.58140.07830.7713-8.785143.0621-70.2225
252.96320.174-0.64971.06660.00333.95920.4010.6731-0.3968-0.1715-0.05420.08020.8531-0.2128-0.22560.791-0.0245-0.17710.5853-0.07730.5614-8.19845.2814-39.7575
268.16830.0744-1.83483.2834-2.98023.04040.55320.2217-0.6306-0.8591-0.35010.24070.79480.0399-0.1530.892-0.063-0.31670.58610.01110.6979-10.106941.2356-36.2304
275.10211.34660.09161.7004-1.0964.4270.26910.6862-0.5498-0.21050.18220.26161.4261-1.1067-0.36761.0959-0.2747-0.20750.89420.03370.7333-19.76942.6677-38.337
283.91521.46284.14431.54911.84154.55770.17290.8362-0.42410.2636-0.00350.61120.5591-0.0505-0.16121.3439-0.381-0.01372.06840.08770.8202-10.4977-4.8654-102.9183
292.66170.3845-0.46312.1378-0.19676.97390.03990.6504-0.15250.11320.00760.07150.9445-0.09730.0010.4012-0.0817-0.07950.93270.06350.5374-10.0503-1.5831-72.8722
305.1298-1.3433-2.52723.92831.1534.75240.12920.3122-0.3663-0.4467-0.22760.5303-0.07060.15870.11550.576-0.0814-0.14581.09260.19660.697-12.8993-4.7894-69.6122
315.14720.2352-0.9692.23371.04356.5047-0.06170.7869-0.33090.0113-0.16970.48680.4814-0.88590.05580.5244-0.1284-0.05831.08330.11710.6662-21.9903-0.7091-71.764
322.8473-0.4938-0.44081.1232-0.06796.1995-0.0505-0.5674-0.1740.27690.12650.33130.1907-0.3976-0.09680.3951-0.05020.14250.75690.03290.68941.166766.8495-81.3356
333.7179-0.8525-0.54721.612-0.44985.6231-0.2905-0.5721-0.12750.52510.32670.7914-0.1997-1.6710.08690.60320.08610.21971.29990.07640.800432.95672.3477-73.2115
347.9524-2.10576.4476.3348-3.62045.8324-0.43620.7021-1.78170.20021.65160.1857-2.2066-1.3879-0.93421.56150.34790.22622.2357-0.02941.018552.905361.33-37.7175
352.63790.62850.182.2001-0.01364.7347-0.0715-0.8663-0.09310.59510.24660.02570.35620.4793-0.02750.56050.14160.03241.0631-0.0450.514954.480764.5605-66.2621
367.94053.8256-3.71737.4144-3.73933.0245-0.0293-0.9034-0.53581.18140.2772-0.0825-0.41390.48550.08930.77560.0322-0.00871.2531-0.08780.615258.027461.9357-69.1934
375.1167-0.92951.42152.43780.02152.73680.0176-0.5210.39270.32130.0685-0.4440.04821.8287-0.17330.74850.0293-0.00391.895-0.20360.725971.678167.7072-71.7189
385.0446-0.33382.09041.5455-0.30732.75170.0721-2.53550.17170.54340.33890.0582-0.11281.2056-0.24170.88720.23750.06921.8684-0.16030.665154.912467.7264-56.9913
Refinement TLS group
IDRefine-IDRefine TLS-IDSelection detailsAuth asym-IDAuth seq-ID
1X-RAY DIFFRACTION1chain 'A' and (resid 2 through 75 )A2 - 75
2X-RAY DIFFRACTION2chain 'A' and (resid 76 through 159 )A76 - 159
3X-RAY DIFFRACTION3chain 'A' and (resid 160 through 273 )A160 - 273
4X-RAY DIFFRACTION4chain 'B' and (resid 2 through 28 )B2 - 28
5X-RAY DIFFRACTION5chain 'B' and (resid 29 through 134 )B29 - 134
6X-RAY DIFFRACTION6chain 'B' and (resid 135 through 159 )B135 - 159
7X-RAY DIFFRACTION7chain 'B' and (resid 160 through 214 )B160 - 214
8X-RAY DIFFRACTION8chain 'B' and (resid 215 through 228 )B215 - 228
9X-RAY DIFFRACTION9chain 'B' and (resid 229 through 273 )B229 - 273
10X-RAY DIFFRACTION10chain 'C' and (resid 2 through 148 )C2 - 148
11X-RAY DIFFRACTION11chain 'C' and (resid 149 through 273 )C149 - 273
12X-RAY DIFFRACTION12chain 'D' and (resid 1 through 28 )D1 - 28
13X-RAY DIFFRACTION13chain 'D' and (resid 29 through 113 )D29 - 113
14X-RAY DIFFRACTION14chain 'D' and (resid 114 through 148 )D114 - 148
15X-RAY DIFFRACTION15chain 'D' and (resid 149 through 189 )D149 - 189
16X-RAY DIFFRACTION16chain 'D' and (resid 190 through 219 )D190 - 219
17X-RAY DIFFRACTION17chain 'D' and (resid 220 through 236 )D220 - 236
18X-RAY DIFFRACTION18chain 'D' and (resid 237 through 273 )D237 - 273
19X-RAY DIFFRACTION19chain 'E' and (resid 1 through 120 )E1 - 120
20X-RAY DIFFRACTION20chain 'E' and (resid 121 through 159 )E121 - 159
21X-RAY DIFFRACTION21chain 'E' and (resid 160 through 273 )E160 - 273
22X-RAY DIFFRACTION22chain 'F' and (resid 1 through 120 )F1 - 120
23X-RAY DIFFRACTION23chain 'F' and (resid 121 through 273 )F121 - 273
24X-RAY DIFFRACTION24chain 'G' and (resid 2 through 28 )G2 - 28
25X-RAY DIFFRACTION25chain 'G' and (resid 29 through 134 )G29 - 134
26X-RAY DIFFRACTION26chain 'G' and (resid 135 through 159 )G135 - 159
27X-RAY DIFFRACTION27chain 'G' and (resid 160 through 273 )G160 - 273
28X-RAY DIFFRACTION28chain 'H' and (resid 3 through 30 )H3 - 30
29X-RAY DIFFRACTION29chain 'H' and (resid 31 through 134 )H31 - 134
30X-RAY DIFFRACTION30chain 'H' and (resid 135 through 159 )H135 - 159
31X-RAY DIFFRACTION31chain 'H' and (resid 160 through 273 )H160 - 273
32X-RAY DIFFRACTION32chain 'I' and (resid 1 through 134 )I1 - 134
33X-RAY DIFFRACTION33chain 'I' and (resid 135 through 273 )I135 - 273
34X-RAY DIFFRACTION34chain 'J' and (resid 5 through 30 )J5 - 30
35X-RAY DIFFRACTION35chain 'J' and (resid 31 through 134 )J31 - 134
36X-RAY DIFFRACTION36chain 'J' and (resid 135 through 159 )J135 - 159
37X-RAY DIFFRACTION37chain 'J' and (resid 160 through 236 )J160 - 236
38X-RAY DIFFRACTION38chain 'J' and (resid 237 through 273 )J237 - 273

+
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