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- PDB-7nit: X-ray structure of a multidomain BbgIII from Bifidobacterium bifidum -

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Open data


ID or keywords:

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Basic information

Entry
Database: PDB / ID: 7nit
TitleX-ray structure of a multidomain BbgIII from Bifidobacterium bifidum
ComponentsBeta-galactosidase
KeywordsHYDROLASE / Bifidobacterium bifidum / lactose hydrolysis / galactooligosaccharides / transgalactosylation
Function / homology
Function and homology information


hydrolase activity, hydrolyzing O-glycosyl compounds / membrane => GO:0016020 / carbohydrate metabolic process
Similarity search - Function
Bacterial Ig-like domain (group 3) / Ig-like domain, bacterial type / Bacterial Ig-like domain / Bacterial Ig-like domain (group 4) / Glycoside hydrolase family 2, domain 5 / Glycoside hydrolase family 2 C-terminal domain 5 / Domain of unknown function DUF4982 / Domain of unknown function (DUF4982) / Invasin/intimin cell-adhesion fragments / Glycoside hydrolase, family 2 ...Bacterial Ig-like domain (group 3) / Ig-like domain, bacterial type / Bacterial Ig-like domain / Bacterial Ig-like domain (group 4) / Glycoside hydrolase family 2, domain 5 / Glycoside hydrolase family 2 C-terminal domain 5 / Domain of unknown function DUF4982 / Domain of unknown function (DUF4982) / Invasin/intimin cell-adhesion fragments / Glycoside hydrolase, family 2 / Glycoside hydrolase family 2, catalytic domain / Glycosyl hydrolases family 2, TIM barrel domain / Glycoside hydrolase, family 2, immunoglobulin-like beta-sandwich / Glycosyl hydrolases family 2, sugar binding domain / Glycosyl hydrolases family 2 / Glycosyl hydrolases family 2, sugar binding domain / Beta-Galactosidase/glucuronidase domain superfamily / Coagulation factors 5/8 type C domain (FA58C) profile. / F5/8 type C domain / Coagulation factor 5/8 C-terminal domain / Galactose-binding-like domain superfamily / Glycoside hydrolase superfamily / Immunoglobulin-like fold
Similarity search - Domain/homology
DUF4982 domain-containing protein
Similarity search - Component
Biological speciesBifidobacterium bifidum (bacteria)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 2.89 Å
AuthorsMoroz, O.V. / Blagova, E. / Lebedev, A.A. / Sanchez Rodriguez, F. / Rigden, D.J. / Tams, J.W. / Wilting, R. / Vester, J.K. / Longhin, E. / Krogh, K.B.R. ...Moroz, O.V. / Blagova, E. / Lebedev, A.A. / Sanchez Rodriguez, F. / Rigden, D.J. / Tams, J.W. / Wilting, R. / Vester, J.K. / Longhin, E. / Krogh, K.B.R. / Pache, R.A. / Davies, G.J. / Wilson, K.S.
CitationJournal: Acta Crystallogr D Struct Biol / Year: 2021
Title: Multitasking in the gut: the X-ray structure of the multidomain BbgIII from Bifidobacterium bifidum offers possible explanations for its alternative functions.
Authors: Moroz, O.V. / Blagova, E. / Lebedev, A.A. / Sanchez Rodriguez, F. / Rigden, D.J. / Tams, J.W. / Wilting, R. / Vester, J.K. / Longhin, E. / Hansen, G.H. / Krogh, K.B.R.M. / Pache, R.A. / ...Authors: Moroz, O.V. / Blagova, E. / Lebedev, A.A. / Sanchez Rodriguez, F. / Rigden, D.J. / Tams, J.W. / Wilting, R. / Vester, J.K. / Longhin, E. / Hansen, G.H. / Krogh, K.B.R.M. / Pache, R.A. / Davies, G.J. / Wilson, K.S.
History
DepositionFeb 14, 2021Deposition site: PDBE / Processing site: PDBE
Revision 1.0Dec 22, 2021Provider: repository / Type: Initial release
Revision 1.1Jan 31, 2024Group: Data collection / Derived calculations / Refinement description
Category: atom_type / chem_comp_atom ...atom_type / chem_comp_atom / chem_comp_bond / pdbx_initial_refinement_model / struct_ncs_dom_lim
Item: _atom_type.pdbx_N_electrons / _atom_type.pdbx_scat_Z ..._atom_type.pdbx_N_electrons / _atom_type.pdbx_scat_Z / _struct_ncs_dom_lim.beg_auth_comp_id / _struct_ncs_dom_lim.beg_label_asym_id / _struct_ncs_dom_lim.beg_label_comp_id / _struct_ncs_dom_lim.beg_label_seq_id / _struct_ncs_dom_lim.end_auth_comp_id / _struct_ncs_dom_lim.end_label_asym_id / _struct_ncs_dom_lim.end_label_comp_id / _struct_ncs_dom_lim.end_label_seq_id

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: Beta-galactosidase
B: Beta-galactosidase
C: Beta-galactosidase
D: Beta-galactosidase
E: Beta-galactosidase
F: Beta-galactosidase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)840,07624
Polymers838,7156
Non-polymers1,36118
Water1086
1
A: Beta-galactosidase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)140,1065
Polymers139,7861
Non-polymers3204
Water181
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
MethodPISA
2
B: Beta-galactosidase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)139,9183
Polymers139,7861
Non-polymers1322
Water181
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
MethodPISA
3
C: Beta-galactosidase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)140,0144
Polymers139,7861
Non-polymers2283
Water181
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
MethodPISA
4
D: Beta-galactosidase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)140,1065
Polymers139,7861
Non-polymers3204
Water181
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
MethodPISA
5
E: Beta-galactosidase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)139,9183
Polymers139,7861
Non-polymers1322
Water181
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
MethodPISA
6
F: Beta-galactosidase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)140,0144
Polymers139,7861
Non-polymers2283
Water181
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
MethodPISA
Unit cell
Length a, b, c (Å)116.947, 130.042, 200.580
Angle α, β, γ (deg.)86.991, 84.830, 83.788
Int Tables number1
Space group name H-MP1
Noncrystallographic symmetry (NCS)NCS domain:
IDEns-IDDetails
11A
21B
32A
42C
53A
63D
74A
84E
95A
105F
116B
126C
137B
147D
158B
168E
179B
189F
1910C
2010D
2111C
2211E
2312C
2412F
2513D
2613E
2714D
2814F
2915E
3015F

NCS domain segments:
Dom-IDComponent-IDEns-IDBeg auth comp-IDBeg label comp-IDEnd auth comp-IDEnd label comp-IDAuth asym-IDLabel asym-IDAuth seq-IDLabel seq-ID
111SERSERTHRTHRAA15 - 128515 - 1285
221SERSERTHRTHRBB15 - 128515 - 1285
332THRTHRTHRTHRAA11 - 128511 - 1285
442THRTHRTHRTHRCC11 - 128511 - 1285
553THRTHRPHEPHEAA11 - 128611 - 1286
663THRTHRPHEPHEDD11 - 128611 - 1286
774SERSERTHRTHRAA15 - 128515 - 1285
884SERSERTHRTHREE15 - 128515 - 1285
995THRTHRTHRTHRAA11 - 128511 - 1285
10105THRTHRTHRTHRFF11 - 128511 - 1285
11116SERSERTHRTHRBB15 - 128515 - 1285
12126SERSERTHRTHRCC15 - 128515 - 1285
13137SERSERPHEPHEBB15 - 128615 - 1286
14147SERSERPHEPHEDD15 - 128615 - 1286
15158SERSERPHEPHEBB15 - 128615 - 1286
16168SERSERPHEPHEEE15 - 128615 - 1286
17179SERSERTHRTHRBB15 - 128515 - 1285
18189SERSERTHRTHRFF15 - 128515 - 1285
191910SERSERPHEPHECC7 - 12867 - 1286
202010SERSERPHEPHEDD7 - 12867 - 1286
212111SERSERTHRTHRCC15 - 128515 - 1285
222211SERSERTHRTHREE15 - 128515 - 1285
232312SERSERTHRTHRCC7 - 12857 - 1285
242412SERSERTHRTHRFF7 - 12857 - 1285
252513SERSERPHEPHEDD15 - 128615 - 1286
262613SERSERPHEPHEEE15 - 128615 - 1286
272714ARGARGPHEPHEDD6 - 12866 - 1286
282814ARGARGPHEPHEFF6 - 12866 - 1286
292915SERSERTHRTHREE15 - 128515 - 1285
303015SERSERTHRTHRFF15 - 128515 - 1285

NCS ensembles :
IDDetails
1Local NCS retraints between domains: 1 2
2Local NCS retraints between domains: 3 4
3Local NCS retraints between domains: 5 6
4Local NCS retraints between domains: 7 8
5Local NCS retraints between domains: 9 10
6Local NCS retraints between domains: 11 12
7Local NCS retraints between domains: 13 14
8Local NCS retraints between domains: 15 16
9Local NCS retraints between domains: 17 18
10Local NCS retraints between domains: 19 20
11Local NCS retraints between domains: 21 22
12Local NCS retraints between domains: 23 24
13Local NCS retraints between domains: 25 26
14Local NCS retraints between domains: 27 28
15Local NCS retraints between domains: 29 30

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Components

#1: Protein
Beta-galactosidase / / DUF4982 domain-containing protein


Mass: 139785.828 Da / Num. of mol.: 6
Source method: isolated from a genetically manipulated source
Details: 33-1336 construct from sequence corresponding to Uniprot entry A0A415C3Q2, starts after 32 amino-acid signal peptide, sequence conflict Glu/Asp (1165 in this entry, 1197 in Uniprot sequence) ...Details: 33-1336 construct from sequence corresponding to Uniprot entry A0A415C3Q2, starts after 32 amino-acid signal peptide, sequence conflict Glu/Asp (1165 in this entry, 1197 in Uniprot sequence) is due to a random mutation that occurred during cloning
Source: (gene. exp.) Bifidobacterium bifidum (bacteria) / Gene: DW137_08020 / Production host: Bacillus licheniformis (bacteria) / References: UniProt: A0A415C3Q2, beta-galactosidase
#2: Chemical
ChemComp-SO4 / SULFATE ION / Sulfate


Mass: 96.063 Da / Num. of mol.: 4 / Source method: obtained synthetically / Formula: SO4
#3: Chemical
ChemComp-GOL / GLYCEROL / GLYCERIN / PROPANE-1,2,3-TRIOL / Glycerol


Mass: 92.094 Da / Num. of mol.: 8 / Source method: obtained synthetically / Formula: C3H8O3
#4: Chemical
ChemComp-CA / CALCIUM ION


Mass: 40.078 Da / Num. of mol.: 6 / Source method: obtained synthetically / Formula: Ca
#5: Water ChemComp-HOH / water / Water


Mass: 18.015 Da / Num. of mol.: 6 / Source method: isolated from a natural source / Formula: H2O
Compound detailsThe electron density for subunits E and F is poor and only interpretable at the level of secondary ...The electron density for subunits E and F is poor and only interpretable at the level of secondary structure elements, but not at the level of individual residues.
Has ligand of interestN

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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: 2.4 Å3/Da / Density % sol: 65.81 %
Crystal growTemperature: 293 K / Method: vapor diffusion, sitting drop / pH: 4.6 / Details: 1.6 M ammonium sulfate, sodium acetate pH 4.6

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Data collection

DiffractionMean temperature: 100 K / Serial crystal experiment: N
Diffraction sourceSource: SYNCHROTRON / Site: Diamond / Beamline: I04 / Wavelength: 0.9795 Å
DetectorType: DECTRIS EIGER2 XE 16M / Detector: PIXEL / Date: Jun 30, 2019
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.9795 Å / Relative weight: 1
ReflectionResolution: 2.89→199.58 Å / Num. obs: 252478 / % possible obs: 96.2 % / Redundancy: 3.6 % / Biso Wilson estimate: 84.7 Å2 / CC1/2: 0.997 / Rmerge(I) obs: 0.087 / Rpim(I) all: 0.087 / Rrim(I) all: 0.123 / Net I/σ(I): 4.9
Reflection shell

Diffraction-ID: 1

Resolution (Å)Redundancy (%)Rmerge(I) obsMean I/σ(I) obsNum. unique obsCC1/2Rpim(I) allRrim(I) allΧ2% possible all
15.82-199.583.50.02115.751080.9990.0210.030.2290.6
2.89-2.943.71.5140.5128110.3551.5142.1410.4798.4

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Processing

Software
NameVersionClassification
autoPROCdata scaling
Aimlessdata reduction
MOLREPphasing
REFMAC5.8.0267refinement
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: 5dmy, 1gof, 2w1s, 2wdc, 3f2z and 4lpl
Resolution: 2.89→199.578 Å / Cor.coef. Fo:Fc: 0.936 / Cor.coef. Fo:Fc free: 0.928 / WRfactor Rfree: 0.25 / WRfactor Rwork: 0.221 / SU B: 23.17 / SU ML: 0.386 / Average fsc free: 0.7789 / Average fsc work: 0.79 / Cross valid method: FREE R-VALUE / ESU R: 1.191 / ESU R Free: 0.355 / Details: Hydrogens have not been used
RfactorNum. reflection% reflection
Rfree0.247 12540 4.976 %
Rwork0.2141 239474 -
all0.216 --
obs-252014 96.234 %
Solvent computationIon probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.2 Å / Solvent model: MASK BULK SOLVENT
Displacement parametersBiso mean: 113.059 Å2
Baniso -1Baniso -2Baniso -3
1--4.589 Å2-1.162 Å21.388 Å2
2--1.185 Å2-5.199 Å2
3---2.947 Å2
Refinement stepCycle: LAST / Resolution: 2.89→199.578 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms55787 0 74 6 55867
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.0060.01257061
X-RAY DIFFRACTIONr_angle_refined_deg1.2591.63777802
X-RAY DIFFRACTIONr_dihedral_angle_1_deg6.91257432
X-RAY DIFFRACTIONr_dihedral_angle_2_deg33.64123.7582650
X-RAY DIFFRACTIONr_dihedral_angle_3_deg16.354158539
X-RAY DIFFRACTIONr_dihedral_angle_4_deg18.94115222
X-RAY DIFFRACTIONr_chiral_restr0.0860.27886
X-RAY DIFFRACTIONr_gen_planes_refined0.0060.0243727
X-RAY DIFFRACTIONr_nbd_refined0.2130.223657
X-RAY DIFFRACTIONr_nbtor_refined0.3020.238665
X-RAY DIFFRACTIONr_xyhbond_nbd_refined0.1220.21756
X-RAY DIFFRACTIONr_symmetry_nbd_refined0.2740.274
X-RAY DIFFRACTIONr_symmetry_xyhbond_nbd_refined0.0990.22
X-RAY DIFFRACTIONr_mcbond_it11.41211.44129803
X-RAY DIFFRACTIONr_mcangle_it16.80817.14537194
X-RAY DIFFRACTIONr_scbond_it12.60311.22827258
X-RAY DIFFRACTIONr_scangle_it17.85416.81240608
X-RAY DIFFRACTIONr_lrange_it23.574213.972242531
X-RAY DIFFRACTIONr_ncsr_local_group_10.0630.0539328
X-RAY DIFFRACTIONr_ncsr_local_group_20.0490.0540161
X-RAY DIFFRACTIONr_ncsr_local_group_30.0490.0540243
X-RAY DIFFRACTIONr_ncsr_local_group_40.0660.0539177
X-RAY DIFFRACTIONr_ncsr_local_group_50.0460.0540114
X-RAY DIFFRACTIONr_ncsr_local_group_60.0620.0539309
X-RAY DIFFRACTIONr_ncsr_local_group_70.0620.0539374
X-RAY DIFFRACTIONr_ncsr_local_group_80.0540.0539490
X-RAY DIFFRACTIONr_ncsr_local_group_90.0620.0539283
X-RAY DIFFRACTIONr_ncsr_local_group_100.0540.0540206
X-RAY DIFFRACTIONr_ncsr_local_group_110.0690.0539050
X-RAY DIFFRACTIONr_ncsr_local_group_120.0560.0539997
X-RAY DIFFRACTIONr_ncsr_local_group_130.0670.0539109
X-RAY DIFFRACTIONr_ncsr_local_group_140.0520.0540047
X-RAY DIFFRACTIONr_ncsr_local_group_150.060.0539232
Refine LS restraints NCS
Ens-IDDom-IDAuth asym-IDRefine-IDTypeRms dev position (Å)Weight position
11AX-RAY DIFFRACTIONLocal ncs0.06350.05013
12BX-RAY DIFFRACTIONLocal ncs0.06350.05013
23AX-RAY DIFFRACTIONLocal ncs0.048790.05013
24CX-RAY DIFFRACTIONLocal ncs0.048790.05013
35AX-RAY DIFFRACTIONLocal ncs0.049050.05013
36DX-RAY DIFFRACTIONLocal ncs0.049050.05013
47AX-RAY DIFFRACTIONLocal ncs0.066460.05013
48EX-RAY DIFFRACTIONLocal ncs0.066460.05013
59AX-RAY DIFFRACTIONLocal ncs0.046130.05013
510FX-RAY DIFFRACTIONLocal ncs0.046130.05013
611BX-RAY DIFFRACTIONLocal ncs0.061870.05013
612CX-RAY DIFFRACTIONLocal ncs0.061870.05013
713BX-RAY DIFFRACTIONLocal ncs0.062060.05013
714DX-RAY DIFFRACTIONLocal ncs0.062060.05013
815BX-RAY DIFFRACTIONLocal ncs0.054310.05013
816EX-RAY DIFFRACTIONLocal ncs0.054310.05013
917BX-RAY DIFFRACTIONLocal ncs0.062230.05013
918FX-RAY DIFFRACTIONLocal ncs0.062230.05013
1019CX-RAY DIFFRACTIONLocal ncs0.053770.05013
1020DX-RAY DIFFRACTIONLocal ncs0.053770.05013
1121CX-RAY DIFFRACTIONLocal ncs0.069020.05013
1122EX-RAY DIFFRACTIONLocal ncs0.069020.05013
1223CX-RAY DIFFRACTIONLocal ncs0.056080.05013
1224FX-RAY DIFFRACTIONLocal ncs0.056080.05013
1325DX-RAY DIFFRACTIONLocal ncs0.066740.05013
1326EX-RAY DIFFRACTIONLocal ncs0.066740.05013
1427DX-RAY DIFFRACTIONLocal ncs0.052380.05013
1428FX-RAY DIFFRACTIONLocal ncs0.052380.05013
1529EX-RAY DIFFRACTIONLocal ncs0.060430.05013
1530FX-RAY DIFFRACTIONLocal ncs0.060430.05013
LS refinement shell

Refine-ID: X-RAY DIFFRACTION / Total num. of bins used: 20

Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkRfactor allNum. reflection allFsc freeFsc work% reflection obs (%)WRfactor Rwork
2.89-2.9650.3959330.388180850.389193300.4860.49698.38590.391
2.965-3.0460.3949150.384176410.385188510.40.41198.43510.386
3.046-3.1350.368980.347172300.348184060.4820.48398.48960.348
3.135-3.2310.3418670.313167240.314178450.6570.67798.57660.309
3.231-3.3370.3038590.28161950.281173170.7550.76898.48130.272
3.337-3.4540.38490.265155870.267166790.7980.81698.54310.257
3.454-3.5840.298350.249151290.251161630.8420.86298.76880.241
3.584-3.7310.294320.24688270.248154570.8410.86359.90170.24
3.731-3.8970.247210.207139940.209149150.9090.92298.65910.201
3.897-4.0870.2287000.189133130.191142240.9230.93498.51660.184
4.087-4.3080.2076630.17127140.172135400.9390.94998.79620.168
4.308-4.5690.1986340.157120070.159127840.9480.95798.88140.157
4.569-4.8840.1845660.15113250.151120350.9570.96298.80350.153
4.884-5.2750.2135450.167104820.17111880.9410.95398.5610.173
5.275-5.7780.235300.18496230.186102470.9340.94599.08270.191
5.778-6.4590.2534270.19688310.19993630.9230.94198.87860.207
6.459-7.4570.2423940.20676400.20881860.9280.93898.14320.223
7.457-9.130.2053870.18664620.18769610.9470.9598.3910.208
9.13-12.8990.2172530.19149670.19253580.9430.95197.42440.22
12.899-199.5780.2981240.31626090.31529300.8810.8793.27650.548

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