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- PDB-3da7: A conformationally strained, circular permutant of barnase -

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ID or keywords:

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Basic information

Entry
Database: PDB / ID: 3da7
TitleA conformationally strained, circular permutant of barnase
Components
  • Barnase circular permutant
  • Barstar
KeywordsPROTEIN BINDING / CIRCULAR PERMUTANT / PROTEIN-PROTEIN COMPLEX / ENDONUCLEASE / Cytoplasm
Function / homology
Function and homology information


Hydrolases; Acting on ester bonds; Endoribonucleases producing 3'-phosphomonoesters / RNA endonuclease activity / RNA binding / extracellular region / cytoplasm
Similarity search - Function
Severin - #20 / Barstar-like / Barstar (barnase inhibitor) / Barstar (barnase inhibitor) / Barstar-like superfamily / Barnase; Chain D / Barnase / Guanine-specific ribonuclease N1/T1/U2 / Severin / ribonuclease ...Severin - #20 / Barstar-like / Barstar (barnase inhibitor) / Barstar (barnase inhibitor) / Barstar-like superfamily / Barnase; Chain D / Barnase / Guanine-specific ribonuclease N1/T1/U2 / Severin / ribonuclease / Ribonuclease/ribotoxin / 2-Layer Sandwich / 3-Layer(aba) Sandwich / Alpha Beta
Similarity search - Domain/homology
Ribonuclease / Barstar
Similarity search - Component
Biological speciesBacillus amyloliquefaciens (bacteria)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / molecular replacement / Resolution: 2.25 Å
AuthorsMitrousis, G. / Butler, J. / Loh, S.N. / Cingolani, G.
CitationJournal: Biochemistry / Year: 2009
Title: Structural and thermodynamic analysis of a conformationally strained circular permutant of barnase.
Authors: Butler, J.S. / Mitrea, D.M. / Mitrousis, G. / Cingolani, G. / Loh, S.N.
History
DepositionMay 28, 2008Deposition site: RCSB / Processing site: RCSB
Revision 1.0Apr 14, 2009Provider: repository / Type: Initial release
Revision 1.1Jul 13, 2011Group: Version format compliance
Revision 1.2Oct 25, 2017Group: Refinement description / Category: software
Revision 1.3Aug 30, 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: Barnase circular permutant
B: Barnase circular permutant
C: Barstar
D: Barstar
E: Barnase circular permutant
F: Barstar
G: Barnase circular permutant
H: Barstar


Theoretical massNumber of molelcules
Total (without water)91,4188
Polymers91,4188
Non-polymers00
Water6,702372
1
A: Barnase circular permutant
D: Barstar


Theoretical massNumber of molelcules
Total (without water)22,8552
Polymers22,8552
Non-polymers00
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
2
B: Barnase circular permutant
C: Barstar


Theoretical massNumber of molelcules
Total (without water)22,8552
Polymers22,8552
Non-polymers00
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
3
E: Barnase circular permutant
F: Barstar


Theoretical massNumber of molelcules
Total (without water)22,8552
Polymers22,8552
Non-polymers00
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
4
G: Barnase circular permutant
H: Barstar


Theoretical massNumber of molelcules
Total (without water)22,8552
Polymers22,8552
Non-polymers00
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Unit cell
Length a, b, c (Å)38.879, 80.237, 81.922
Angle α, β, γ (deg.)88.100, 76.980, 79.470
Int Tables number1
Space group name H-MP1

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Components

#1: Protein
Barnase circular permutant


Mass: 12501.869 Da / Num. of mol.: 4
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Bacillus amyloliquefaciens (bacteria) / Production host: Escherichia coli (E. coli) / References: UniProt: P00648*PLUS
#2: Protein
Barstar / Ribonuclease inhibitor


Mass: 10352.739 Da / Num. of mol.: 4
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Bacillus amyloliquefaciens (bacteria) / Production host: Escherichia coli (E. coli) / References: UniProt: P11540
#3: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 372 / 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: 2.68 Å3/Da / Density % sol: 54.06 %
Crystal growTemperature: 277 K / Method: vapor diffusion, hanging drop / pH: 8.5
Details: 1M Tris, 14% peg 8k, 150mM (NH4)2SO4, pH 8.5, VAPOR DIFFUSION, HANGING DROP, temperature 277K

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: CHESS / Beamline: A1 / Wavelength: 0.972 Å
DetectorType: ADSC QUANTUM 210 / Detector: CCD / Date: May 10, 2006
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.972 Å / Relative weight: 1
ReflectionResolution: 2.15→30 Å / Num. obs: 50783 / % possible obs: 96 % / Redundancy: 3 % / Rmerge(I) obs: 0.109 / Χ2: 6.279 / Net I/σ(I): 17
Reflection shell
Resolution (Å)Redundancy (%)Rmerge(I) obsNum. unique allΧ2% possible all
2.15-2.232.20.39250575.94495.2
2.23-2.322.60.41850328.26195
2.32-2.4230.42151009.23696.5
2.42-2.553.40.38651268.45297.4
2.55-2.713.50.29651507.10297.4
2.71-2.923.50.19451635.27797.6
2.92-3.213.40.11951714.59197.5
3.21-3.673.10.07451214.60697.2
3.67-4.632.90.05549654.71294.1
4.63-302.90.0548984.73992.5

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Phasing

PhasingMethod: molecular replacement

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Processing

Software
NameVersionClassificationNB
DENZOdata reduction
SCALEPACKdata scaling
MOLREPphasing
REFMACrefinement
PDB_EXTRACT3.005data extraction
ADSCQuantumdata collection
HKL-2000data reduction
HKL-2000data scaling
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: pdb entry 1brs
Resolution: 2.25→19.87 Å / Cor.coef. Fo:Fc: 0.935 / Cor.coef. Fo:Fc free: 0.899 / SU B: 12.107 / SU ML: 0.159 / Cross valid method: THROUGHOUT / σ(F): 0 / ESU R: 0.323 / ESU R Free: 0.234 / Stereochemistry target values: Engh & Huber
RfactorNum. reflection% reflectionSelection details
Rfree0.25 2198 5.1 %RANDOM
Rwork0.205 ---
obs0.207 42697 94.93 %-
Solvent computationIon probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.2 Å / Solvent model: MASK
Displacement parametersBiso mean: 37.599 Å2
Baniso -1Baniso -2Baniso -3
1-0.03 Å20.01 Å2-0.02 Å2
2--0.15 Å20.02 Å2
3----0.18 Å2
Refinement stepCycle: LAST / Resolution: 2.25→19.87 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms6173 0 0 372 6545
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.010.0226303
X-RAY DIFFRACTIONr_angle_refined_deg1.0641.9478534
X-RAY DIFFRACTIONr_dihedral_angle_1_deg15.1425763
X-RAY DIFFRACTIONr_dihedral_angle_2_deg36.67324.433300
X-RAY DIFFRACTIONr_dihedral_angle_3_deg15.474151081
X-RAY DIFFRACTIONr_dihedral_angle_4_deg19.091535
X-RAY DIFFRACTIONr_chiral_restr0.0820.2926
X-RAY DIFFRACTIONr_gen_planes_refined0.0040.024774
X-RAY DIFFRACTIONr_nbd_refined0.250.23132
X-RAY DIFFRACTIONr_nbtor_refined0.3250.24383
X-RAY DIFFRACTIONr_xyhbond_nbd_refined0.1890.2471
X-RAY DIFFRACTIONr_symmetry_vdw_refined0.3280.280
X-RAY DIFFRACTIONr_symmetry_hbond_refined0.2440.224
X-RAY DIFFRACTIONr_mcbond_it2.0931.53825
X-RAY DIFFRACTIONr_mcangle_it2.49126119
X-RAY DIFFRACTIONr_scbond_it3.16232478
X-RAY DIFFRACTIONr_scangle_it4.8224.52415
LS refinement shellResolution: 2.25→2.308 Å / Total num. of bins used: 20
RfactorNum. reflection% reflection
Rfree0.29 146 -
Rwork0.238 2960 -
all-3106 -
obs--93.61 %
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)
12.4018-0.63431.31850.7578-0.41353.66860.12380.0164-0.01180.0014-0.18890.01850.19670.07340.06510.03230.00610.02070.02570.04690.054-0.8612-6.01287.79
212.7414-7.7819-17.06479.86470.852740.77020.5080.03490.3942-1.25080.1963-0.9185-1.0365-1.215-0.70430.2876-0.1049-0.0857-0.05460.02340.109-3.0677-20.388715.0934
33.7976-0.0683-0.93672.2892-2.6956.43690.023-0.2083-0.1045-0.0706-0.1387-0.11240.28830.34320.11580.03310.1350.03350.08020.08130.0517.6107-12.221210.8987
44.4326-7.6412-6.772729.074914.072617.34260.57180.40340.9143-1.7268-1.0324-2.2824-0.69220.270.46050.01840.2770.24750.15650.19470.075314.4043-5.1021-4.9004
51.7347-0.6647-0.13392.3066-0.41152.4550.2387-0.00370.1378-0.3253-0.2625-0.31560.13560.31980.02380.10410.08190.02650.07340.05960.00944.5267-3.56270.8602
62.19951.0795-1.90021.6495-1.17425.25420.0576-0.00450.06450.05070.03880.0972-0.3348-0.0266-0.09640.0410.00640.022-0.018-0.00470.0607-18.982139.5801-46.8457
76.51283.7767-2.5722.259-0.89296.2088-0.07220.45270.10810.03490.08780.0469-0.7592-0.1312-0.01560.16970.0107-0.0095-0.04780.0260.0428-16.088247.4942-53.1831
81.79361.7489-0.03974.5385-3.63986.74250.1937-0.1697-0.15680.2-0.416-0.1931-0.39140.35620.22230.0838-0.091-0.01370.03880.04160.0393-7.650644.0588-47.8911
94.28715.01742.494510.37891.15927.12250.8396-0.3864-0.23751.4301-0.945-0.6698-0.63160.49910.10540.3514-0.3703-0.15890.17330.1093-0.0257-7.802741.8082-32.6114
107.80890.80292.85762.0439-1.97987.9462-0.0030.57640.155-0.01550.0912-0.0390.556-0.3092-0.08820.0607-0.06170.02240.065-0.00080.0343-20.984329.9443-49.1938
116.12660.68470.47732.2654-0.48062.4963-0.0412-0.23140.1907-0.00590.1320.0052-0.2071-0.4923-0.0907-0.00670.00460.03130.0316-0.01890.1058-30.079423.6838-27.8838
121.87021.7178-2.62161.7949-1.35838.75060.0477-0.02910.05450.1832-0.03840.1391-0.3475-0.246-0.0093-0.01380.05550.01280.05690.02190.0975-26.333633.1913-31.2148
131.4483-1.34880.20672.7436-2.38643.32660.02560.00840.0460.1187-0.00010.0098-0.32260.1651-0.02550.0336-0.03460.0210.0179-0.0030.0751-17.22730.3471-30.9947
1414.49283.25598.17334.52611.04618.76790.00150.0131-0.4373-0.06370.12930.1875-0.0443-0.1389-0.1307-0.03290.00910.0322-0.0111-0.00880.0984-29.587915.2465-30.9448
153.9644-0.8991-0.82630.72380.18323.03720.04480.11320.1392-0.1836-0.1061-0.09240.02330.00220.06140.0128-0.00630.01410.00790.00110.0807-17.728320.5334-33.4127
1623.8969-7.245-12.36723.16742.53217.927-0.17181.20030.43250.10870.2614-0.23030.1014-0.7447-0.0896-0.0209-0.0027-0.02140.0404-0.03620.0804-8.183813.4257-14.7175
172.2242-0.58510.01921.6794-0.34131.0440.15060.0255-0.0270.0249-0.06260.07120.099-0.1486-0.0880.0454-0.0127-0.00450.04060.02370.0542-7.23524.3629-5.7035
186.3972-2.835-0.74313.7418-0.63280.45870.07490.10090.117-0.0826-0.00060.18940.1146-0.0245-0.07440.00160.0063-0.02160.0216-0.02490.0635-4.772414.1215-9.7786
1914.92570.81041.43220.1534-0.35711.86550.2736-0.46090.2140.2528-0.17490.0201-0.0571-0.0642-0.09870.08150.01040.01670.01860.01960.0399-1.254914.2223-1.4587
206.0845-3.7711.60664.9455-2.51023.8913-0.01730.27450.06340.3796-0.2726-0.08440.0040.24910.28990.01040.0199-0.02280.0382-0.02230.05654.778214.2708-9.2431
211.88380.60371.53711.27920.30684.39620.07710.0082-0.01070.1047-0.0566-0.01230.2695-0.1035-0.02050.0729-0.0430.0174-0.0109-0.02570.0467-3.097566.5985-44.2546
223.29331.7691.54063.14120.93514.3748-0.06460.23190.0914-0.18930.09230.01510.27880.1593-0.02760.0797-0.0191-0.0251-0.0372-0.01160.0277-1.331962.7763-49.9455
231.81180.0115-0.68033.86869.626136.02360.1223-0.3842-0.18750.2217-0.55510.51891.2407-1.87520.43280.0343-0.18120.03540.1893-0.00050.1183-13.632663.8374-41.3416
241.91950.7236-1.82627.13921.944212.57460.2631-0.2773-0.01791.0675-0.31490.59221.0889-0.90630.05180.2152-0.31570.11040.2222-0.0104-0.0031-11.25662.7778-28.1065
250.41930.09741.53333.42091.04285.7459-0.2132-0.01740.4089-0.26230.01960.1875-0.16620.08270.19360.0033-0.0434-0.00620.01360.01170.0819-2.305475.2503-46.0296
261.0110.6620.06812.83460.96031.95590.0568-0.039-0.04170.11050.0245-0.13830.26820.1147-0.08120.01190.005-0.01560.0176-0.00850.07165.313778.6823-29.6498
2713.23488.63312.67438.15064.43743.4190.0154-0.5879-0.25210.5951-0.2825-0.35790.52730.17650.26710.0098-0.02280.04040.02260.0060.0887-3.874282.5466-23.3608
282.4263-0.8248-0.08034.96573.20255.34080.22990.16580.1516-0.25050.0482-0.2296-0.31310.0891-0.2781-0.0206-0.0101-0.0170.0397-0.00450.11649.20392.2641-32.5617
2917.16430.7749-2.09792.3289-0.57160.35560.22460.3215-0.0388-0.1669-0.29160.1421-0.34-0.1110.06690.0380.00260.01040.0376-0.07040.0454-3.440485.6442-35.5921
301.39091.06922.09396.9537-0.493.87250.0154-0.04050.05120.1965-0.10810.0723-0.0703-0.16840.0927-0.06630.00560.01970.0533-0.04450.1027-3.814489.0346-26.5189
3110.3274-0.30298.9848.30689.983620.46930.2793-0.8498-0.48910.8305-0.21590.49551.8399-1.8272-0.06340.0923-0.12150.02790.18040.00440.1405-3.432237.897116.1474
320.63280.15210.27870.8145-1.41434.3828-0.01760.01240.09050.1426-0.0494-0.0757-0.6376-0.15030.0670.10010.0263-0.0022-0.0049-0.030.0753.460648.86435.871
3321.8158-0.0193-1.846312.8701-1.37548.8208-0.5604-1.2220.87311.14650.5923-0.5189-2.2622-1.784-0.03190.30980.3457-0.08320.2299-0.12250.0446-5.977155.288513.1532
342.86393.31032.92658.67310.812824.2819-0.38690.19760.0731-0.00910.00280.6441-0.6318-0.81590.3841-0.00890.17120.00520.0952-0.02050.0755-8.904549.3807-1.4815
352.0961-0.67440.59871.2002-0.57423.92240.1136-0.0457-0.0223-0.1609-0.04980.2146-0.1574-0.3301-0.06390.04030.0671-0.02020.0158-0.04760.0478-2.124545.5579-1.9753
3613.0964-12.57396.546412.1163-6.59015.3798-0.13690.1655-0.4963-0.25510.38620.1854-0.36880.2398-0.2493-0.01370.01990.02880.00580.03740.13910.578227.7418-15.5209
371.6975-2.1720.83373.65130.42222.95110.15270.0830.1965-0.2786-0.0625-0.3134-0.14230.3929-0.0902-0.0014-0.02430.04320.0666-0.00420.09913.908536.246-8.4156
381.4593-0.1533-0.38242.15770.43510.69430.06420.00510.0756-0.0691-0.07930.1399-0.1140.03050.01520.02360.02910.01680.03450.00960.05462.86135.3646-8.3206
3912.3515-2.18727.68690.5179-1.42259.52050.15630.2043-0.33670.10170.18560.01440.51590.3633-0.342-0.030.02940.0280.0525-0.01170.111514.302520.5064-7.0771
403.6578-0.91770.3971.95210.42113.4941-0.0588-0.12440.16370.0491-0.02270.06140.1017-0.05050.08150.02220.01170.0128-0.0116-0.02520.07262.507527.1881-5.3529
Refinement TLS group
IDRefine-IDRefine TLS-IDAuth asym-IDLabel asym-IDAuth seq-IDLabel seq-ID
1X-RAY DIFFRACTION1AA4 - 453 - 44
2X-RAY DIFFRACTION2AA46 - 5145 - 50
3X-RAY DIFFRACTION3AA52 - 7351 - 72
4X-RAY DIFFRACTION4AA74 - 8073 - 79
5X-RAY DIFFRACTION5AA81 - 11280 - 111
6X-RAY DIFFRACTION6BB3 - 422 - 41
7X-RAY DIFFRACTION7BB43 - 5942 - 58
8X-RAY DIFFRACTION8BB60 - 7459 - 73
9X-RAY DIFFRACTION9BB75 - 10074 - 99
10X-RAY DIFFRACTION10BB101 - 111100 - 110
11X-RAY DIFFRACTION11CC1 - 201 - 20
12X-RAY DIFFRACTION12CC21 - 3521 - 35
13X-RAY DIFFRACTION13CC36 - 5236 - 52
14X-RAY DIFFRACTION14CC53 - 6553 - 65
15X-RAY DIFFRACTION15CC66 - 9066 - 90
16X-RAY DIFFRACTION16DD2 - 102 - 10
17X-RAY DIFFRACTION17DD11 - 4411 - 44
18X-RAY DIFFRACTION18DD45 - 6445 - 64
19X-RAY DIFFRACTION19DD65 - 7965 - 79
20X-RAY DIFFRACTION20DD80 - 9080 - 90
21X-RAY DIFFRACTION21EE3 - 372 - 36
22X-RAY DIFFRACTION22EE38 - 6237 - 61
23X-RAY DIFFRACTION23EE63 - 7762 - 76
24X-RAY DIFFRACTION24EE78 - 9677 - 95
25X-RAY DIFFRACTION25EE97 - 11296 - 111
26X-RAY DIFFRACTION26FF1 - 461 - 46
27X-RAY DIFFRACTION27FF47 - 5347 - 53
28X-RAY DIFFRACTION28FF54 - 6754 - 67
29X-RAY DIFFRACTION29FF68 - 8268 - 82
30X-RAY DIFFRACTION30FF83 - 9083 - 90
31X-RAY DIFFRACTION31GG2 - 61 - 5
32X-RAY DIFFRACTION32GG7 - 466 - 45
33X-RAY DIFFRACTION33GG56 - 6555 - 64
34X-RAY DIFFRACTION34GG66 - 7865 - 77
35X-RAY DIFFRACTION35GG79 - 11278 - 111
36X-RAY DIFFRACTION36HH3 - 103 - 10
37X-RAY DIFFRACTION37HH11 - 3411 - 34
38X-RAY DIFFRACTION38HH35 - 5535 - 55
39X-RAY DIFFRACTION39HH56 - 6456 - 64
40X-RAY DIFFRACTION40HH - P65 - 9165 - 1

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  • In this update, many items about electron microscopy experimental information are reorganized (e.g. em_software).
  • Now, EM Navigator and Yorodumi are based on the updated data.

External links:wwPDB Remediation / Enriched Model Files Conforming to OneDep Data Standards Now Available in the PDB FTP Archive

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Yorodumi

Thousand views of thousand structures

  • Yorodumi is a browser for structure data from EMDB, PDB, SASBDB, etc.
  • This page is also the successor to EM Navigator detail page, and also detail information page/front-end page for Omokage search.
  • The word "yorodu" (or yorozu) is an old Japanese word meaning "ten thousand". "mi" (miru) is to see.

Related info.:EMDB / PDB / SASBDB / Comparison of 3 databanks / Yorodumi Search / Aug 31, 2016. New EM Navigator & Yorodumi / Yorodumi Papers / Jmol/JSmol / Function and homology information / Changes in new EM Navigator and Yorodumi

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