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- PDB-5f73: Crystal structure of Mutant S12T of Adenosine/Methylthioadenosine... -

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Basic information

Entry
Database: PDB / ID: 5f73
TitleCrystal structure of Mutant S12T of Adenosine/Methylthioadenosine Phosphorylase in APO form
ComponentsMethylthioadenosine phosphorylase
KeywordsTRANSFERASE
Function / homology
Function and homology information


S-methyl-5'-thioadenosine phosphorylase / S-methyl-5-thioadenosine phosphorylase activity / L-methionine salvage from methylthioadenosine / purine ribonucleoside salvage / nucleus / cytosol
Similarity search - Function
Methylthioadenosine phosphorylase (MTAP) / Purine phosphorylase, family 2, conserved site / Purine and other phosphorylases family 2 signature. / Nucleoside phosphorylase domain / Nucleoside phosphorylase domain / Phosphorylase superfamily / Nucleoside phosphorylase superfamily / Rossmann fold / 3-Layer(aba) Sandwich / Alpha Beta
Similarity search - Domain/homology
S-methyl-5'-thioadenosine phosphorylase
Similarity search - Component
Biological speciesSchistosoma mansoni (invertebrata)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / molecular replacement / Resolution: 2.06 Å
AuthorsTorini, J.R.S. / Brandao-Neto, J. / DeMarco, R. / Pereira, H.M.
Funding support Brazil, 2items
OrganizationGrant numberCountry
Sao Paulo Research Foundation (FAPESP)2012/14223-9 Brazil
Brazilian National Council for Scientific and Technological Development (CNPq)474402/2013-4 Brazil
CitationJournal: PLoS Negl Trop Dis / Year: 2016
Title: Crystal Structure of Schistosoma mansoni Adenosine Phosphorylase/5'-Methylthioadenosine Phosphorylase and Its Importance on Adenosine Salvage Pathway.
Authors: Torini, J.R. / Brandao-Neto, J. / DeMarco, R. / Pereira, H.D.
History
DepositionDec 7, 2015Deposition site: RCSB / Processing site: RCSB
Revision 1.0Dec 14, 2016Provider: repository / Type: Initial release
Revision 1.1Dec 21, 2016Group: Database references
Revision 1.2Sep 27, 2017Group: Author supporting evidence / Data collection / Category: diffrn_source / pdbx_audit_support
Item: _diffrn_source.pdbx_synchrotron_site / _pdbx_audit_support.funding_organization
Revision 1.3Nov 1, 2017Group: Author supporting evidence / Category: pdbx_struct_assembly_auth_evidence
Revision 1.4Apr 17, 2019Group: Author supporting evidence / Data collection / Category: pdbx_audit_support / Item: _pdbx_audit_support.funding_organization
Revision 1.5Jan 1, 2020Group: Author supporting evidence / Category: pdbx_audit_support / Item: _pdbx_audit_support.funding_organization
Revision 1.6Sep 27, 2023Group: Data collection / Database references / Refinement description
Category: chem_comp_atom / chem_comp_bond ...chem_comp_atom / chem_comp_bond / database_2 / pdbx_initial_refinement_model
Item: _database_2.pdbx_DOI / _database_2.pdbx_database_accession

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: Methylthioadenosine phosphorylase
B: Methylthioadenosine phosphorylase
C: Methylthioadenosine phosphorylase
D: Methylthioadenosine phosphorylase
E: Methylthioadenosine phosphorylase
F: Methylthioadenosine phosphorylase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)212,21712
Polymers211,6416
Non-polymers5766
Water18,2311012
1
A: Methylthioadenosine phosphorylase
B: Methylthioadenosine phosphorylase
C: Methylthioadenosine phosphorylase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)106,1096
Polymers105,8213
Non-polymers2883
Water543
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area7280 Å2
ΔGint-100 kcal/mol
Surface area33220 Å2
MethodPISA
2
D: Methylthioadenosine phosphorylase
E: Methylthioadenosine phosphorylase
F: Methylthioadenosine phosphorylase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)106,1096
Polymers105,8213
Non-polymers2883
Water543
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area7460 Å2
ΔGint-96 kcal/mol
Surface area33070 Å2
MethodPISA
Unit cell
Length a, b, c (Å)81.440, 82.160, 150.910
Angle α, β, γ (deg.)90.000, 101.500, 90.000
Int Tables number4
Space group name H-MP1211
DetailsTrimer confirmed by gel filtration

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Components

#1: Protein
Methylthioadenosine phosphorylase


Mass: 35273.508 Da / Num. of mol.: 6 / Fragment: Enzyme / Mutation: S12T
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Schistosoma mansoni (invertebrata) / Plasmid: pET28a / Production host: Escherichia coli (E. coli) / Strain (production host): BL21(DE3)
References: UniProt: I0B503, S-methyl-5'-thioadenosine phosphorylase
#2: Chemical
ChemComp-SO4 / SULFATE ION


Mass: 96.063 Da / Num. of mol.: 6 / Source method: obtained synthetically / Formula: SO4
#3: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 1012 / 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: 2.34 Å3/Da / Density % sol: 47.4 %
Crystal growTemperature: 291 K / Method: vapor diffusion, hanging drop / pH: 6.5
Details: 100mM Bris-tris or Mes, 14-18% PEG3350, pH 6.5, VAPOR DIFFUSION, HANGING DROP, temperature 291K
PH range: 6.3-6.6

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: Diamond / Beamline: I04-1 / Wavelength: 0.92 Å
DetectorType: DECTRIS PILATUS 2M / Detector: PIXEL / Date: Nov 5, 2013
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.92 Å / Relative weight: 1
ReflectionResolution: 2.06→39.58 Å / Num. obs: 119129 / % possible obs: 98.8 % / Redundancy: 4.2 % / Biso Wilson estimate: 25.92 Å2 / Rmerge(I) obs: 0.094 / Net I/σ(I): 11.2
Reflection shell
Resolution (Å)Redundancy (%)Rmerge(I) obsDiffraction-IDRejects% possible all
2.06-2.113.80.5951094.3
9.21-39.584.30.031098.3

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Phasing

PhasingMethod: molecular replacement

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Processing

Software
NameVersionClassification
xia2data scaling
PHASERphasing
PHENIXrefinement
PDB_EXTRACT3.15data extraction
xia2data reduction
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: 4L5Y
Resolution: 2.06→39.58 Å / SU ML: 0.23 / Cross valid method: FREE R-VALUE / σ(F): 1.34 / Phase error: 23.4 / Stereochemistry target values: ML
RfactorNum. reflection% reflection
Rfree0.2162 6071 5.03 %
Rwork0.1753 114716 -
obs0.1773 120787 98.59 %
Solvent computationShrinkage radii: 0.9 Å / VDW probe radii: 1.11 Å / Solvent model: FLAT BULK SOLVENT MODEL
Displacement parametersBiso max: 131.59 Å2 / Biso mean: 34.3295 Å2 / Biso min: 14.82 Å2
Refinement stepCycle: final / Resolution: 2.06→39.58 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms12757 0 30 1012 13799
Biso mean--25.13 38.81 -
Num. residues----1692
Refine LS restraints
Refine-IDTypeDev idealNumber
X-RAY DIFFRACTIONf_bond_d0.00713042
X-RAY DIFFRACTIONf_angle_d0.79617726
X-RAY DIFFRACTIONf_chiral_restr0.0532074
X-RAY DIFFRACTIONf_plane_restr0.0052274
X-RAY DIFFRACTIONf_dihedral_angle_d11.8697803
LS refinement shell

Refine-ID: X-RAY DIFFRACTION / Total num. of bins used: 30

Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkNum. reflection all% reflection obs (%)
2.06-2.07330.34572100.26753688389896
2.0733-2.09770.26861700.25153643381393
2.0977-2.12330.2771870.22593566375394
2.1233-2.15020.26282290.21873825405499
2.1502-2.17850.2491830.209638634046100
2.1785-2.20830.27881790.200738914070100
2.2083-2.23990.2371760.200638444020100
2.2399-2.27330.261930.20943852404599
2.2733-2.30880.24092050.191938254030100
2.3088-2.34670.23942000.188238804080100
2.3467-2.38720.23152230.186738244047100
2.3872-2.43060.24062150.187438324047100
2.4306-2.47730.24142010.187838784079100
2.4773-2.52790.22752180.181338534071100
2.5279-2.58290.22672260.174238014027100
2.5829-2.64290.20272140.17083852406699
2.6429-2.7090.24642160.16933801401799
2.709-2.78230.19691980.17273830402899
2.7823-2.86420.22882020.17273840404299
2.8642-2.95660.23621800.18983829400998
2.9566-3.06230.24492110.1953643385494
3.0623-3.18490.23942150.19293827404299
3.1849-3.32990.22942010.182539044105100
3.3299-3.50540.20752210.175938454066100
3.5054-3.7250.20292040.167139064110100
3.725-4.01270.17351910.15183854404599
4.0127-4.41640.1821770.13583889406699
4.4164-5.05540.15952040.13233881408599
5.0554-6.36890.20412120.16763781399397
6.3689-79.86560.1892100.17013969417999
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)
11.4981-0.4847-0.40140.9264-0.4690.56990.2065-0.52870.02040.6419-0.0820.63980.3576-0.546-0.0850.3898-0.09680.17560.45940.02980.558-97.9589-23.667974.4565
20.85770.1815-0.02231.6425-0.56341.476-0.0381-0.014-0.09270.1431-0.04090.30630.3061-0.27340.07570.274-0.04260.10020.25460.03740.4002-91.497-26.993268.9855
31.8614-0.5732-0.91232.3380.5593.10350.02820.1504-0.2191-0.0831-0.09190.07670.1473-0.13460.07660.1739-0.00080.00480.20790.01630.2504-84.9185-18.984557.0894
41.80550.86840.463.66230.85282.667-0.1737-0.0970.20370.25750.09560.7274-0.3654-0.49670.06680.23850.11950.02050.36750.04980.3745-101.0957-5.495559.9139
51.16930.65210.25960.63590.36561.29230.05040.18040.03360.00790.06270.0335-0.2127-0.1019-0.09930.24650.04880.05910.18010.03850.2456-83.708-9.63161.1565
60.9091-0.271-0.43492.80822.75435.15040.1815-0.1653-0.22910.04850.6413-1.0663-0.48780.6749-0.78250.35660.00240.05340.3171-0.04420.5323-83.9381-6.580570.9867
72.6608-0.2454-0.5391.7592-0.21372.5303-0.0056-0.0885-0.12310.15230.0530.43020.0766-0.4223-0.05050.22550.00290.04360.28920.060.3871-96.8831-17.796763.4723
80.5608-1.5008-0.26844.30641.53493.213-0.31610.3853-0.37760.01880.02330.35870.4782-0.16040.34210.2585-0.04090.01880.3172-0.03270.2755-66.8256-37.561630.3483
90.72580.59251.61323.71023.1086.6823-0.26870.0593-0.47410.211-0.04-0.22180.86050.12840.25680.4392-0.01930.09670.2284-0.00470.4896-63.7326-43.703143.0254
101.4632-0.2996-0.88841.03860.35091.8973-0.05630.204-0.27510.065-0.06130.09650.23-0.13570.11830.27630.00680.02980.2117-0.05420.2626-65.9226-35.553337.7644
111.5581-0.21090.12170.92070.55411.8140.0670.22730.1166-0.142-0.10830.056-0.2589-0.17250.03880.31170.02840.03040.2261-0.00680.2342-66.728-19.15735.7331
121.7868-0.6513-0.59440.83520.89130.95980.09370.07320.0209-0.0153-0.08970.0812-0.0578-0.1794-0.01770.28470.02980.0420.2359-0.00040.2279-73.9844-18.435844.8397
130.21210.2597-0.30581.07370.01350.6546-0.1481-0.3130.11870.19210.07090.482-0.2259-0.45670.12180.33210.11080.04190.4729-0.05780.3775-77.7114-23.424836.0663
147.55830.6124-1.30151.41010.26122.3668-0.0220.7202-0.1956-0.2371-0.20750.3218-0.0064-0.70490.24480.31980.0273-0.03380.4491-0.14780.3365-76.6001-28.660125.6799
150.841-0.01380.41641.60331.56352.54270.06050.1854-0.0686-0.15420.1367-0.1979-0.09041.3447-0.18670.24850.02280.04750.3486-0.01520.2742-47.7233-25.782538.3184
162.16460.29122.71961.465-1.47796.04130.26060.0762-0.18380.0065-0.1914-0.52920.61930.8029-0.06870.2994-0.00410.00450.23870.03660.3076-44.1715-14.660972.9451
170.93580.1675-0.11421.943-0.41910.64570.0236-0.076800.084-0.0189-0.0044-0.06480.0495-0.00540.2464-0.00560.02590.1935-0.00160.1985-55.8493-12.35569.5336
180.4682-0.22-0.36930.29550.56671.21570.04130.0470.1069-0.1068-0.03410.0455-0.1667-0.0119-0.02790.27030.00150.0150.16750.01750.2251-59.4331-7.444457.7432
194.9380.5669-3.63262.3729-1.0726.0599-0.02440.8498-0.0257-0.3410.1152-0.18130.2546-0.5753-0.11920.291-0.0460.04960.3010.00780.2416-51.0209-8.4857.7481
202.1726-0.467-1.73062.48211.62799.23780.0650.07250.0935-0.13140.062-0.50180.03540.4197-0.14440.2693-0.05010.02090.20920.02190.3076-41.6082-6.123163.5024
217.01332.93691.72612.81860.79421.81220.2752-0.7851-0.14220.7832-0.22560.14880.1139-0.2608-0.01830.61470.04170.16140.30050.00530.2664-65.2997-8.24983.9425
222.0858-0.7094-0.20392.44010.38431.4497-0.01490.1062-0.2535-0.06-0.0641-0.18590.19810.19910.06930.220.02830.03820.2298-0.00510.2892-85.911112.03254.5285
231.82990.66470.03911.9732-0.81722.67960.071-0.23570.07120.1795-0.14960.1158-0.13240.13380.08680.1671-0.01710.01910.2156-0.02970.2301-92.702425.689714.1317
242.2320.6321-1.95943.8336-1.15011.8076-0.1823-0.0647-0.5603-0.0795-0.122-0.19750.31560.05850.30450.2598-0.0018-0.00560.28770.04320.3067-91.692610.702122.5852
251.6187-0.3109-0.13781.2153-0.15991.05120.0157-0.1190.14580.0897-0.0098-0.1963-0.17460.2252-0.01030.2044-0.0413-0.01710.2389-0.02220.2079-87.251930.45313.4965
260.39890.0825-0.46970.4688-0.00750.60360.32960.2873-0.0309-0.28110.23090.6983-0.2345-0.454-0.13980.2427-0.0183-0.04560.32380.07860.3963-93.029132.53271.8702
272.6251.0931-0.7831.08320.25792.1672-0.036-0.0101-0.29170.0877-0.1451-0.4712-0.11920.5530.13070.1670.0484-0.01890.244-0.02930.2928-79.289719.052412.1702
280.08510.5337-0.07833.5799-1.60942.4642-0.1354-0.6597-0.6883-0.0525-0.2048-0.33450.15480.38920.2940.27680.00020.04190.36650.16210.3485-110.68182.639144.8468
290.99140.1755-0.26431.24670.08151.5989-0.0996-0.1575-0.30270.0389-0.02430.07230.22370.02910.09880.3155-0.03350.05520.24020.07860.3069-111.88442.912136.2149
303.51020.44490.53361.5160.11521.40580.1215-0.33310.05740.2188-0.1129-0.0783-0.42710.10060.0090.4059-0.05330.05040.30040.0330.2349-102.363824.263338.9619
311.92130.3717-0.0181.3532-0.60022.54740.0901-0.36430.04180.2714-0.15630.0692-0.19520.1330.05840.3227-0.02340.08230.25220.040.2805-111.689320.21239.515
321.23940.5887-0.68940.6913-0.27561.28860.0469-0.09150.02650.2454-0.00540.1606-0.04640.0639-0.01810.2379-0.01570.02520.19740.02650.1793-102.707521.35229.9001
330.2246-0.4740.34791.3054-0.3820.7708-0.23750.37130.0850.09890.0076-0.3363-0.15960.53330.24340.3788-0.09030.03250.58490.09370.3411-98.714616.412138.4389
346.2426-0.4064-1.92991.01260.42891.73440.1589-0.9456-0.23660.4868-0.3418-0.211-0.12240.74420.16340.3676-0.0515-0.02870.58870.230.3604-101.326111.092949.1917
350.86510.2409-0.70791.56-1.31861.447-0.0175-0.18820.0080.02440.05890.3530.0726-1.2115-0.07530.3271-0.01230.11440.4428-0.0020.4911-129.257215.387836.8018
360.9424-1.1762-0.87092.00851.19821.7653-0.11340.4502-0.2704-0.2169-0.28460.42270.8994-0.7540.10730.3650.0023-0.05520.3892-0.10980.3093-129.548123.23410.7277
371.7653-0.3206-0.58731.60720.60310.8360.09610.1936-0.1296-0.1014-0.13140.139-0.0575-0.18770.01870.19530.0258-0.04480.2028-0.02280.1795-121.037128.99186.2592
380.809-0.2092-0.07320.5827-0.02471.04570.0277-0.05280.01590.0822-0.05010.0786-0.1056-0.1280.01450.21560.00830.01020.1709-0.00340.2194-118.811433.062617.2507
392.5804-0.9592-0.41482.10130.34771.34480.1530.37240.1495-0.1826-0.12530.1296-0.1326-0.1948-0.02230.26160.0172-0.06060.1734-0.04240.2013-124.234533.52432.5273
Refinement TLS group
IDRefine-IDRefine TLS-IDSelection detailsAuth asym-IDAuth seq-ID
1X-RAY DIFFRACTION1chain 'A' and (resid 4 through 26 )A0
2X-RAY DIFFRACTION2chain 'A' and (resid 27 through 90 )A0
3X-RAY DIFFRACTION3chain 'A' and (resid 91 through 139 )A0
4X-RAY DIFFRACTION4chain 'A' and (resid 140 through 164 )A0
5X-RAY DIFFRACTION5chain 'A' and (resid 165 through 220 )A0
6X-RAY DIFFRACTION6chain 'A' and (resid 221 through 238 )A0
7X-RAY DIFFRACTION7chain 'A' and (resid 239 through 291 )A0
8X-RAY DIFFRACTION8chain 'B' and (resid 2 through 23 )B0
9X-RAY DIFFRACTION9chain 'B' and (resid 24 through 38 )B0
10X-RAY DIFFRACTION10chain 'B' and (resid 39 through 90 )B0
11X-RAY DIFFRACTION11chain 'B' and (resid 91 through 182 )B0
12X-RAY DIFFRACTION12chain 'B' and (resid 183 through 217 )B0
13X-RAY DIFFRACTION13chain 'B' and (resid 218 through 241 )B0
14X-RAY DIFFRACTION14chain 'B' and (resid 242 through 269 )B0
15X-RAY DIFFRACTION15chain 'B' and (resid 270 through 291 )B0
16X-RAY DIFFRACTION16chain 'C' and (resid 3 through 23 )C0
17X-RAY DIFFRACTION17chain 'C' and (resid 24 through 164 )C0
18X-RAY DIFFRACTION18chain 'C' and (resid 165 through 220 )C0
19X-RAY DIFFRACTION19chain 'C' and (resid 221 through 242 )C0
20X-RAY DIFFRACTION20chain 'C' and (resid 243 through 269 )C0
21X-RAY DIFFRACTION21chain 'C' and (resid 270 through 291 )C0
22X-RAY DIFFRACTION22chain 'D' and (resid 3 through 89 )D0
23X-RAY DIFFRACTION23chain 'D' and (resid 90 through 120 )D0
24X-RAY DIFFRACTION24chain 'D' and (resid 121 through 139 )D0
25X-RAY DIFFRACTION25chain 'D' and (resid 140 through 220 )D0
26X-RAY DIFFRACTION26chain 'D' and (resid 221 through 242 )D0
27X-RAY DIFFRACTION27chain 'D' and (resid 243 through 291 )D0
28X-RAY DIFFRACTION28chain 'E' and (resid 2 through 23 )E0
29X-RAY DIFFRACTION29chain 'E' and (resid 24 through 90 )E0
30X-RAY DIFFRACTION30chain 'E' and (resid 91 through 110 )E0
31X-RAY DIFFRACTION31chain 'E' and (resid 111 through 182 )E0
32X-RAY DIFFRACTION32chain 'E' and (resid 183 through 217 )E0
33X-RAY DIFFRACTION33chain 'E' and (resid 218 through 241 )E0
34X-RAY DIFFRACTION34chain 'E' and (resid 242 through 270 )E0
35X-RAY DIFFRACTION35chain 'E' and (resid 271 through 291 )E0
36X-RAY DIFFRACTION36chain 'F' and (resid 3 through 29 )F0
37X-RAY DIFFRACTION37chain 'F' and (resid 30 through 164 )F0
38X-RAY DIFFRACTION38chain 'F' and (resid 165 through 242 )F0
39X-RAY DIFFRACTION39chain 'F' and (resid 243 through 292 )F0

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