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- PDB-6qqo: Aplysia californica AChBP in complex with 2-Fluoro-(carbamoylpyri... -

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Basic information

Entry
Database: PDB / ID: 6qqo
TitleAplysia californica AChBP in complex with 2-Fluoro-(carbamoylpyridinyl)deschloroepibatidine analogue (3)
ComponentsSoluble acetylcholine receptor
KeywordsSIGNALING PROTEIN / AChBP / Acetylcholine
Function / homology
Function and homology information


extracellular ligand-gated monoatomic ion channel activity / transmembrane signaling receptor activity / membrane / identical protein binding / metal ion binding
Similarity search - Function
Acetylcholine Binding Protein; Chain: A, / Neurotransmitter-gated ion-channel ligand-binding domain / Neurotransmitter-gated ion-channel / Neurotransmitter-gated ion-channel ligand-binding domain / Neurotransmitter-gated ion-channel ligand-binding domain superfamily / Neurotransmitter-gated ion-channel ligand binding domain / Distorted Sandwich / Mainly Beta
Similarity search - Domain/homology
Chem-JC8 / PHOSPHATE ION / Soluble acetylcholine receptor
Similarity search - Component
Biological speciesAplysia californica (California sea hare)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 2.5 Å
AuthorsBueno, R.V. / Davis, S. / Dawson, A. / Hunter, W.N.
CitationJournal: Acta Crystallogr.,Sect.D / Year: 2022
Title: Interactions between 2'-fluoro-(carbamoylpyridinyl)deschloroepibatidine analogues and acetylcholine-binding protein inform on potent antagonist activity against nicotinic receptors
Authors: Bueno, R.V. / Davis, S. / Dawson, A. / Ondachi, P.W. / Carroll, F.I. / Hunter, W.N.
History
DepositionFeb 18, 2019Deposition site: PDBE / Processing site: PDBE
Revision 1.0Mar 18, 2020Provider: repository / Type: Initial release
Revision 1.1Jul 29, 2020Group: Data collection / Derived calculations / Structure summary
Category: chem_comp / entity ...chem_comp / entity / pdbx_chem_comp_identifier / pdbx_entity_nonpoly / struct_conn / struct_site / struct_site_gen
Item: _chem_comp.name / _entity.pdbx_description ..._chem_comp.name / _entity.pdbx_description / _pdbx_entity_nonpoly.name / _struct_conn.pdbx_role
Description: Carbohydrate remediation / Provider: repository / Type: Remediation
Revision 1.2Mar 2, 2022Group: Database references / Structure summary
Category: chem_comp / citation ...chem_comp / citation / citation_author / database_2
Item: _chem_comp.pdbx_synonyms / _citation.country ..._chem_comp.pdbx_synonyms / _citation.country / _citation.journal_abbrev / _citation.journal_id_ASTM / _citation.journal_id_CSD / _citation.journal_id_ISSN / _citation.journal_volume / _citation.pdbx_database_id_DOI / _citation.title / _citation.year / _database_2.pdbx_DOI / _database_2.pdbx_database_accession
Revision 1.3Jan 24, 2024Group: Data collection / Refinement description
Category: chem_comp_atom / chem_comp_bond ...chem_comp_atom / chem_comp_bond / pdbx_initial_refinement_model / struct_ncs_dom_lim
Item: _struct_ncs_dom_lim.beg_auth_comp_id / _struct_ncs_dom_lim.beg_label_asym_id ..._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: Soluble acetylcholine receptor
B: Soluble acetylcholine receptor
C: Soluble acetylcholine receptor
D: Soluble acetylcholine receptor
E: Soluble acetylcholine receptor
F: Soluble acetylcholine receptor
G: Soluble acetylcholine receptor
H: Soluble acetylcholine receptor
I: Soluble acetylcholine receptor
J: Soluble acetylcholine receptor
hetero molecules


Theoretical massNumber of molelcules
Total (without water)292,63487
Polymers283,26810
Non-polymers9,36777
Water11,872659
1
A: Soluble acetylcholine receptor
B: Soluble acetylcholine receptor
C: Soluble acetylcholine receptor
D: Soluble acetylcholine receptor
E: Soluble acetylcholine receptor
hetero molecules


Theoretical massNumber of molelcules
Total (without water)146,45646
Polymers141,6345
Non-polymers4,82241
Water905
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area23180 Å2
ΔGint-21 kcal/mol
Surface area40630 Å2
MethodPISA
2
F: Soluble acetylcholine receptor
G: Soluble acetylcholine receptor
H: Soluble acetylcholine receptor
I: Soluble acetylcholine receptor
J: Soluble acetylcholine receptor
hetero molecules


Theoretical massNumber of molelcules
Total (without water)146,17841
Polymers141,6345
Non-polymers4,54536
Water905
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area22210 Å2
ΔGint-54 kcal/mol
Surface area40680 Å2
MethodPISA
Unit cell
Length a, b, c (Å)209.826, 134.060, 131.719
Angle α, β, γ (deg.)90.00, 102.56, 90.00
Int Tables number5
Space group name H-MC121
Noncrystallographic symmetry (NCS)NCS domain:
IDEns-IDDetails
11A
21B
12A
22C
13A
23D
14A
24E
15A
25F
16A
26G
17A
27H
18A
28I
19A
29J
110B
210C
111B
211D
112B
212E
113B
213F
114B
214G
115B
215H
116B
216I
117B
217J
118C
218D
119C
219E
120C
220F
121C
221G
122C
222H
123C
223I
124C
224J
125D
225E
126D
226F
127D
227G
128D
228H
129D
229I
130D
230J
131E
231F
132E
232G
133E
233H
134E
234I
135E
235J
136F
236G
137F
237H
138F
238I
139F
239J
140G
240H
141G
241I
142G
242J
143H
243I
144H
244J
145I
245J

NCS domain segments:

Component-ID: 0 / Beg auth comp-ID: GLN / Beg label comp-ID: GLN / Refine code: 0

Dom-IDEns-IDEnd auth comp-IDEnd label comp-IDAuth asym-IDLabel asym-IDAuth seq-IDLabel seq-ID
11ARGARGAA20 - 22420 - 224
21ARGARGBB20 - 22420 - 224
12ARGARGAA20 - 22420 - 224
22ARGARGCC20 - 22420 - 224
13GLUGLUAA20 - 22320 - 223
23GLUGLUDD20 - 22320 - 223
14GLUGLUAA20 - 22320 - 223
24GLUGLUEE20 - 22320 - 223
15ARGARGAA20 - 22420 - 224
25ARGARGFF20 - 22420 - 224
16GLUGLUAA20 - 22320 - 223
26GLUGLUGG20 - 22320 - 223
17ARGARGAA20 - 22420 - 224
27ARGARGHH20 - 22420 - 224
18ARGARGAA20 - 22420 - 224
28ARGARGII20 - 22420 - 224
19ARGARGAA20 - 22420 - 224
29ARGARGJJ20 - 22420 - 224
110ARGARGBB20 - 22420 - 224
210ARGARGCC20 - 22420 - 224
111GLUGLUBB20 - 22320 - 223
211GLUGLUDD20 - 22320 - 223
112GLUGLUBB20 - 22320 - 223
212GLUGLUEE20 - 22320 - 223
113ARGARGBB20 - 22420 - 224
213ARGARGFF20 - 22420 - 224
114GLUGLUBB20 - 22320 - 223
214GLUGLUGG20 - 22320 - 223
115ARGARGBB20 - 22420 - 224
215ARGARGHH20 - 22420 - 224
116ARGARGBB20 - 22420 - 224
216ARGARGII20 - 22420 - 224
117ARGARGBB20 - 22420 - 224
217ARGARGJJ20 - 22420 - 224
118GLUGLUCC20 - 22320 - 223
218GLUGLUDD20 - 22320 - 223
119GLUGLUCC20 - 22320 - 223
219GLUGLUEE20 - 22320 - 223
120ARGARGCC20 - 22420 - 224
220ARGARGFF20 - 22420 - 224
121GLUGLUCC20 - 22320 - 223
221GLUGLUGG20 - 22320 - 223
122ARGARGCC20 - 22420 - 224
222ARGARGHH20 - 22420 - 224
123ARGARGCC20 - 22420 - 224
223ARGARGII20 - 22420 - 224
124ARGARGCC20 - 22420 - 224
224ARGARGJJ20 - 22420 - 224
125ARGARGDD20 - 22520 - 225
225ARGARGEE20 - 22520 - 225
126GLUGLUDD20 - 22320 - 223
226GLUGLUFF20 - 22320 - 223
127ARGARGDD20 - 22520 - 225
227ARGARGGG20 - 22520 - 225
128GLUGLUDD20 - 22320 - 223
228GLUGLUHH20 - 22320 - 223
129GLUGLUDD20 - 22320 - 223
229GLUGLUII20 - 22320 - 223
130GLUGLUDD20 - 22320 - 223
230GLUGLUJJ20 - 22320 - 223
131GLUGLUEE20 - 22320 - 223
231GLUGLUFF20 - 22320 - 223
132ARGARGEE20 - 22520 - 225
232ARGARGGG20 - 22520 - 225
133GLUGLUEE20 - 22320 - 223
233GLUGLUHH20 - 22320 - 223
134GLUGLUEE20 - 22320 - 223
234GLUGLUII20 - 22320 - 223
135GLUGLUEE20 - 22320 - 223
235GLUGLUJJ20 - 22320 - 223
136GLUGLUFF20 - 22320 - 223
236GLUGLUGG20 - 22320 - 223
137ARGARGFF20 - 22420 - 224
237ARGARGHH20 - 22420 - 224
138ARGARGFF20 - 22420 - 224
238ARGARGII20 - 22420 - 224
139ARGARGFF20 - 22420 - 224
239ARGARGJJ20 - 22420 - 224
140GLUGLUGG20 - 22320 - 223
240GLUGLUHH20 - 22320 - 223
141GLUGLUGG20 - 22320 - 223
241GLUGLUII20 - 22320 - 223
142GLUGLUGG20 - 22320 - 223
242GLUGLUJJ20 - 22320 - 223
143ARGARGHH20 - 22420 - 224
243ARGARGII20 - 22420 - 224
144ARGARGHH20 - 22420 - 224
244ARGARGJJ20 - 22420 - 224
145ARGARGII20 - 22420 - 224
245ARGARGJJ20 - 22420 - 224

NCS ensembles :
ID
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45

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Components

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Protein / Sugars , 2 types, 20 molecules ABCDEFGHIJ

#1: Protein
Soluble acetylcholine receptor


Mass: 28326.750 Da / Num. of mol.: 10
Source method: isolated from a genetically manipulated source
Details: The sequence conflicts are because we have used the AChBP gene sequence provided in the Aplysia californica genome (http://genome-euro.ucsc.edu/cgi-bin/hgGateway?hgsid=589127391_ ...Details: The sequence conflicts are because we have used the AChBP gene sequence provided in the Aplysia californica genome (http://genome-euro.ucsc.edu/cgi-bin/hgGateway?hgsid=589127391_t0Vv5jEv8KyWGt7MjPEaZKNxTL3u&redirect=manual&source=genome.cse.ucsc.edu), which differs from the UniProt Q8WSF8 Sequence by identifying both ALA 60 and and ALA 155 as VAL.
Source: (gene. exp.) Aplysia californica (California sea hare)
Cell line (production host): Sf9 / Production host: Spodoptera frugiperda (fall armyworm) / References: UniProt: Q8WSF8
#5: Sugar
ChemComp-NAG / 2-acetamido-2-deoxy-beta-D-glucopyranose / N-acetyl-beta-D-glucosamine / 2-acetamido-2-deoxy-beta-D-glucose / 2-acetamido-2-deoxy-D-glucose / 2-acetamido-2-deoxy-glucose / N-ACETYL-D-GLUCOSAMINE / N-Acetylglucosamine


Type: D-saccharide, beta linking / Mass: 221.208 Da / Num. of mol.: 10
Source method: isolated from a genetically manipulated source
Formula: C8H15NO6
IdentifierTypeProgram
DGlcpNAcbCONDENSED IUPAC CARBOHYDRATE SYMBOLGMML 1.0
N-acetyl-b-D-glucopyranosamineCOMMON NAMEGMML 1.0
b-D-GlcpNAcIUPAC CARBOHYDRATE SYMBOLPDB-CARE 1.0
GlcNAcSNFG CARBOHYDRATE SYMBOLGMML 1.0

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Non-polymers , 4 types, 726 molecules

#2: Chemical
ChemComp-JC8 / 6-[5-[(1~{R},2~{R},4~{S})-7-azabicyclo[2.2.1]heptan-2-yl]-2-fluoranyl-pyridin-3-yl]pyridine-3-carboxamide


Mass: 312.341 Da / Num. of mol.: 10 / Source method: obtained synthetically / Formula: C17H17FN4O / Feature type: SUBJECT OF INVESTIGATION
#3: Chemical
ChemComp-PO4 / PHOSPHATE ION / Phosphate


Mass: 94.971 Da / Num. of mol.: 15 / Source method: obtained synthetically / Formula: PO4
#4: Chemical...
ChemComp-EDO / 1,2-ETHANEDIOL / ETHYLENE GLYCOL / Ethylene glycol


Mass: 62.068 Da / Num. of mol.: 42 / Source method: obtained synthetically / Formula: C2H6O2
#6: Water ChemComp-HOH / water / Water


Mass: 18.015 Da / Num. of mol.: 659 / 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: 3.49 Å3/Da / Density % sol: 64.79 % / Description: Block
Crystal growTemperature: 293 K / Method: vapor diffusion, hanging drop / pH: 4.2
Details: Reservoir: 0.2 M NaCl 0.1 M Phosphate/citrate pH 4.2 10% PEG 8000 Buffer: 50 mM Tris 250 mM NaCl Protein concentration 4 mg/ml Microseeded

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Data collection

DiffractionMean temperature: 100 K / Serial crystal experiment: N
Diffraction sourceSource: SYNCHROTRON / Site: Diamond / Beamline: I24 / Wavelength: 0.96858 Å
DetectorType: DECTRIS PILATUS 6M-F / Detector: PIXEL / Date: Jul 3, 2017
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.96858 Å / Relative weight: 1
ReflectionResolution: 2.5→46.4 Å / Num. obs: 109552 / % possible obs: 89.7 % / Redundancy: 2.3 % / Biso Wilson estimate: 13.6 Å2 / CC1/2: 0.93 / Rmerge(I) obs: 0.118 / Net I/σ(I): 6.9
Reflection shellResolution: 2.5→2.54 Å / Redundancy: 2.3 % / Rmerge(I) obs: 0.559 / Mean I/σ(I) obs: 2.6 / Num. unique obs: 12673 / CC1/2: 0.46 / % possible all: 91.8

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Processing

Software
NameVersionClassification
REFMAC5.8.0189refinement
XDSdata reduction
Aimlessdata scaling
PHASERphasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: 5o87
Resolution: 2.5→46.4 Å / Cor.coef. Fo:Fc: 0.92 / Cor.coef. Fo:Fc free: 0.902 / SU B: 7.279 / SU ML: 0.155 / Cross valid method: THROUGHOUT / ESU R: 0.372 / ESU R Free: 0.231 / Details: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS
RfactorNum. reflection% reflectionSelection details
Rfree0.21161 5421 4.9 %RANDOM
Rwork0.1928 ---
obs0.19372 104105 89.09 %-
Solvent computationIon probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.2 Å
Displacement parametersBiso mean: 20.005 Å2
Baniso -1Baniso -2Baniso -3
1--2.7 Å2-0 Å2-0.44 Å2
2--1.62 Å2-0 Å2
3---1.16 Å2
Refinement stepCycle: LAST / Resolution: 2.5→46.4 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms16381 0 613 659 17653
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.0110.01917477
X-RAY DIFFRACTIONr_bond_other_d0.0020.0215419
X-RAY DIFFRACTIONr_angle_refined_deg1.5611.96523838
X-RAY DIFFRACTIONr_angle_other_deg0.933.00135898
X-RAY DIFFRACTIONr_dihedral_angle_1_deg7.9615.0172069
X-RAY DIFFRACTIONr_dihedral_angle_2_deg31.51324.481790
X-RAY DIFFRACTIONr_dihedral_angle_3_deg12.681152731
X-RAY DIFFRACTIONr_dihedral_angle_4_deg18.0871593
X-RAY DIFFRACTIONr_chiral_restr0.0830.22654
X-RAY DIFFRACTIONr_gen_planes_refined0.0060.02119819
X-RAY DIFFRACTIONr_gen_planes_other0.0010.023498
X-RAY DIFFRACTIONr_nbd_refined
X-RAY DIFFRACTIONr_nbd_other
X-RAY DIFFRACTIONr_nbtor_refined
X-RAY DIFFRACTIONr_nbtor_other
X-RAY DIFFRACTIONr_xyhbond_nbd_refined
X-RAY DIFFRACTIONr_xyhbond_nbd_other
X-RAY DIFFRACTIONr_metal_ion_refined
X-RAY DIFFRACTIONr_metal_ion_other
X-RAY DIFFRACTIONr_symmetry_vdw_refined
X-RAY DIFFRACTIONr_symmetry_vdw_other
X-RAY DIFFRACTIONr_symmetry_hbond_refined
X-RAY DIFFRACTIONr_symmetry_hbond_other
X-RAY DIFFRACTIONr_symmetry_metal_ion_refined
X-RAY DIFFRACTIONr_symmetry_metal_ion_other
X-RAY DIFFRACTIONr_mcbond_it1.6031.888278
X-RAY DIFFRACTIONr_mcbond_other1.6031.888275
X-RAY DIFFRACTIONr_mcangle_it2.8262.8110335
X-RAY DIFFRACTIONr_mcangle_other2.8262.8110335
X-RAY DIFFRACTIONr_scbond_it2.2782.2439198
X-RAY DIFFRACTIONr_scbond_other2.2782.2449199
X-RAY DIFFRACTIONr_scangle_it
X-RAY DIFFRACTIONr_scangle_other3.8313.26213502
X-RAY DIFFRACTIONr_long_range_B_refined6.39221.58617917
X-RAY DIFFRACTIONr_long_range_B_other6.37221.58417865
X-RAY DIFFRACTIONr_rigid_bond_restr
X-RAY DIFFRACTIONr_sphericity_free
X-RAY DIFFRACTIONr_sphericity_bonded
Refine LS restraints NCS

Refine-ID: X-RAY DIFFRACTION / Type: interatomic distance / Weight position: 0.05

Ens-IDDom-IDAuth asym-IDNumberRms dev position (Å)
11A129260.03
12B129260.03
21A129500.03
22C129500.03
31A127420.03
32D127420.03
41A127540.04
42E127540.04
51A129300.03
52F129300.03
61A128200.02
62G128200.02
71A128960.04
72H128960.04
81A129040.04
82I129040.04
91A128840.03
92J128840.03
101B129580.02
102C129580.02
111B127100.03
112D127100.03
121B127580.03
122E127580.03
131B129340.03
132F129340.03
141B127980.02
142G127980.02
151B128540.04
152H128540.04
161B129340.03
162I129340.03
171B129000.03
172J129000.03
181C127520.03
182D127520.03
191C127660.04
192E127660.04
201C129640.02
202F129640.02
211C128400.02
212G128400.02
221C128800.04
222H128800.04
231C129380.03
232I129380.03
241C129320.03
242J129320.03
251D128160.05
252E128160.05
261D127280.04
262F127280.04
271D128720.04
272G128720.04
281D126960.04
282H126960.04
291D127440.03
292I127440.03
301D127460.03
302J127460.03
311E127420.04
312F127420.04
321E128740.05
322G128740.05
331E126960.04
332H126960.04
341E127240.05
342I127240.05
351E127480.04
352J127480.04
361F128200.03
362G128200.03
371F128940.04
372H128940.04
381F129140.03
382I129140.03
391F129240.03
392J129240.03
401G127900.04
402H127900.04
411G128020.04
412I128020.04
421G128080.03
422J128080.03
431H129360.04
432I129360.04
441H129200.04
442J129200.04
451I128920.02
452J128920.02
LS refinement shellResolution: 2.5→2.565 Å / Total num. of bins used: 20
RfactorNum. reflection% reflection
Rfree0.286 416 -
Rwork0.274 7813 -
obs--91.2 %

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