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- PDB-2j3z: Crystal structure of the enzymatic component C2-I of the C2-toxin... -

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Basic information

Entry
Database: PDB / ID: 2j3z
TitleCrystal structure of the enzymatic component C2-I of the C2-toxin from Clostridium botulinum at pH 6.1
ComponentsC2 TOXIN COMPONENT I
KeywordsTOXIN / ADP-RIBOSYLTRANSFERASE / CLOSTRIDIUM BOTULINUM
Function / homology
Function and homology information


extracellular region
Similarity search - Function
Binary exotoxin A, clostridial type / Toxin ADP-ribosyltransferase; Chain A, domain 1 / Toxin ADP-ribosyltransferase; Chain A, domain 1 / ADP ribosyltransferase / ADP-ribosyltransferase exoenzyme / Toxin-related mono-ADP-ribosyltransferase (TR mART) core domain profile. / Alpha-Beta Complex / Alpha Beta
Similarity search - Domain/homology
: / C2 toxin (Component I)
Similarity search - Component
Biological speciesCLOSTRIDIUM BOTULINUM (bacteria)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 2.3 Å
AuthorsSchleberger, C. / Hochmann, H. / Barth, H. / Aktories, K. / Schulz, G.E.
CitationJournal: J.Mol.Biol. / Year: 2006
Title: Structure and Action of the Binary C2 Toxin from Clostridium Botulinum.
Authors: Schleberger, C. / Hochmann, H. / Barth, H. / Aktories, K. / Schulz, G.E.
History
DepositionAug 23, 2006Deposition site: PDBE / Processing site: PDBE
Revision 1.0Oct 11, 2006Provider: repository / Type: Initial release
Revision 1.1May 8, 2011Group: Version format compliance
Revision 1.2Jul 13, 2011Group: Version format compliance
Revision 1.3May 8, 2019Group: Data collection / Experimental preparation / Other
Category: database_PDB_rev / database_PDB_rev_record ...database_PDB_rev / database_PDB_rev_record / exptl_crystal_grow / pdbx_database_proc / pdbx_database_status
Item: _exptl_crystal_grow.method / _pdbx_database_status.recvd_author_approval
Revision 1.4Jul 24, 2019Group: Data collection / Category: diffrn_source / Item: _diffrn_source.pdbx_synchrotron_site
Revision 1.5Dec 13, 2023Group: Data collection / Database references ...Data collection / Database references / Derived calculations / Other / Refinement description
Category: chem_comp_atom / chem_comp_bond ...chem_comp_atom / chem_comp_bond / database_2 / pdbx_database_status / pdbx_initial_refinement_model / pdbx_struct_conn_angle / struct_conn / struct_ncs_dom_lim
Item: _database_2.pdbx_DOI / _database_2.pdbx_database_accession ..._database_2.pdbx_DOI / _database_2.pdbx_database_accession / _pdbx_database_status.status_code_sf / _pdbx_struct_conn_angle.ptnr1_auth_asym_id / _pdbx_struct_conn_angle.ptnr1_auth_comp_id / _pdbx_struct_conn_angle.ptnr1_auth_seq_id / _pdbx_struct_conn_angle.ptnr1_label_asym_id / _pdbx_struct_conn_angle.ptnr1_label_atom_id / _pdbx_struct_conn_angle.ptnr1_label_comp_id / _pdbx_struct_conn_angle.ptnr1_label_seq_id / _pdbx_struct_conn_angle.ptnr3_auth_asym_id / _pdbx_struct_conn_angle.ptnr3_auth_comp_id / _pdbx_struct_conn_angle.ptnr3_auth_seq_id / _pdbx_struct_conn_angle.ptnr3_label_asym_id / _pdbx_struct_conn_angle.ptnr3_label_atom_id / _pdbx_struct_conn_angle.ptnr3_label_comp_id / _pdbx_struct_conn_angle.ptnr3_label_seq_id / _pdbx_struct_conn_angle.value / _struct_conn.pdbx_dist_value / _struct_conn.ptnr1_auth_asym_id / _struct_conn.ptnr1_auth_comp_id / _struct_conn.ptnr1_auth_seq_id / _struct_conn.ptnr1_label_asym_id / _struct_conn.ptnr1_label_atom_id / _struct_conn.ptnr1_label_comp_id / _struct_conn.ptnr1_label_seq_id / _struct_conn.ptnr2_auth_asym_id / _struct_conn.ptnr2_auth_comp_id / _struct_conn.ptnr2_auth_seq_id / _struct_conn.ptnr2_label_asym_id / _struct_conn.ptnr2_label_atom_id / _struct_conn.ptnr2_label_comp_id / _struct_conn.ptnr2_label_seq_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
Remark 650 HELIX DETERMINATION METHOD: AUTHOR PROVIDED.
Remark 700 SHEET DETERMINATION METHOD: AUTHOR PROVIDED.

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: C2 TOXIN COMPONENT I
B: C2 TOXIN COMPONENT I
C: C2 TOXIN COMPONENT I
D: C2 TOXIN COMPONENT I
E: C2 TOXIN COMPONENT I
F: C2 TOXIN COMPONENT I
hetero molecules


Theoretical massNumber of molelcules
Total (without water)301,71065
Polymers296,3506
Non-polymers5,36059
Water10,665592
1
A: C2 TOXIN COMPONENT I
hetero molecules


Theoretical massNumber of molelcules
Total (without water)50,29911
Polymers49,3921
Non-polymers90810
Water181
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
MethodPQS
2
B: C2 TOXIN COMPONENT I
hetero molecules


Theoretical massNumber of molelcules
Total (without water)50,46213
Polymers49,3921
Non-polymers1,07112
Water181
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
MethodPQS
3
C: C2 TOXIN COMPONENT I
hetero molecules


Theoretical massNumber of molelcules
Total (without water)50,30711
Polymers49,3921
Non-polymers91610
Water181
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
MethodPQS
4
D: C2 TOXIN COMPONENT I
hetero molecules


Theoretical massNumber of molelcules
Total (without water)50,21110
Polymers49,3921
Non-polymers8199
Water181
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
MethodPQS
5
E: C2 TOXIN COMPONENT I
hetero molecules


Theoretical massNumber of molelcules
Total (without water)50,21510
Polymers49,3921
Non-polymers8239
Water181
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
MethodPQS
6
F: C2 TOXIN COMPONENT I
hetero molecules


Theoretical massNumber of molelcules
Total (without water)50,21510
Polymers49,3921
Non-polymers8239
Water181
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
MethodPQS
Unit cell
Length a, b, c (Å)93.421, 272.898, 246.290
Angle α, β, γ (deg.)90.00, 90.00, 90.00
Int Tables number20
Space group name H-MC2221
Noncrystallographic symmetry (NCS)NCS domain:
IDEns-IDDetails
11A
21B
31C
12A
22B
32C
13D
23E
33F
14D
24E
34F

NCS domain segments:

Component-ID: 1 / Refine code: 4

Dom-IDEns-IDBeg auth comp-IDBeg label comp-IDEnd auth comp-IDEnd label comp-IDAuth asym-IDLabel asym-IDAuth seq-IDLabel seq-ID
11PHEPHEPHEPHEAA10 - 22110 - 221
21PHEPHEPHEPHEBB10 - 22110 - 221
31PHEPHEPHEPHECC10 - 22110 - 221
12TYRTYRVALVALAA222 - 420222 - 420
22TYRTYRVALVALBB222 - 420222 - 420
32TYRTYRVALVALCC222 - 420222 - 420
13PHEPHEPHEPHEDD10 - 22110 - 221
23PHEPHEPHEPHEEE10 - 22110 - 221
33PHEPHEPHEPHEFF10 - 22110 - 221
14TYRTYRVALVALDD222 - 420222 - 420
24TYRTYRVALVALEE222 - 420222 - 420
34TYRTYRVALVALFF222 - 420222 - 420

NCS ensembles :
ID
1
2
3
4

NCS oper:
IDCodeMatrixVector
1given(0.999969, 0.007882, 0.000791), (0.003409, -0.518299, 0.855193), (0.00715, -0.855163, -0.518309)-1.0429, 112.2725, 185.6765
2given(0.999966, -0.002033, -0.007963), (-0.007922, -0.495986, -0.868294), (-0.002184, 0.868328, -0.495986)0.6703, 215.8952, -1.8538
3given(0.999958, -0.002249, 0.008904), (-0.008827, -0.502976, 0.864255), (0.002535, -0.864298, -0.502975)-0.4305, 110.0379, 186.6458
4given(0.999365, -0.033918, -0.01088), (-0.025332, -0.462005, -0.886516), (0.025042, 0.886229, -0.462571)3.8825, 215.4, -6.8478

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Components

#1: Protein
C2 TOXIN COMPONENT I / ADP-RIBOSYLTRANSFERASE C2-I


Mass: 49391.684 Da / Num. of mol.: 6 / Mutation: YES
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) CLOSTRIDIUM BOTULINUM (bacteria) / Plasmid: PET-3B / Production host: ESCHERICHIA COLI (E. coli) / Strain (production host): BL21 / References: UniProt: O69275
#2: Chemical
ChemComp-CO / COBALT (II) ION


Mass: 58.933 Da / Num. of mol.: 7 / Source method: obtained synthetically / Formula: Co
#3: Chemical
ChemComp-GOL / GLYCEROL / GLYCERIN / PROPANE-1,2,3-TRIOL / Glycerol


Mass: 92.094 Da / Num. of mol.: 12 / Source method: obtained synthetically / Formula: C3H8O3
#4: Chemical...
ChemComp-SO4 / SULFATE ION / Sulfate


Mass: 96.063 Da / Num. of mol.: 40 / Source method: obtained synthetically / Formula: SO4
#5: Water ChemComp-HOH / water / Water


Mass: 18.015 Da / Num. of mol.: 592 / Source method: isolated from a natural source / Formula: H2O
Compound detailsENGINEERED RESIDUE IN CHAIN A, HIS 77 TO ARG ENGINEERED RESIDUE IN CHAIN B, HIS 77 TO ARG ...ENGINEERED RESIDUE IN CHAIN A, HIS 77 TO ARG ENGINEERED RESIDUE IN CHAIN B, HIS 77 TO ARG ENGINEERED RESIDUE IN CHAIN C, HIS 77 TO ARG ENGINEERED RESIDUE IN CHAIN D, HIS 77 TO ARG ENGINEERED RESIDUE IN CHAIN E, HIS 77 TO ARG ENGINEERED RESIDUE IN CHAIN F, HIS 77 TO ARG

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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: 2.66 Å3/Da / Density % sol: 53.8 %
Crystal growMethod: vapor diffusion, hanging drop / pH: 6.1
Details: PROTEIN: 14 MG/ML IN H2O RESERVOIR: 0.1 M MES PH 6.1, 2.1 M (NH4)2SO4, 0.03 M COCL2 HANGING DROP 1:1

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: EMBL/DESY, HAMBURG / Beamline: X11 / Wavelength: 0.813
DetectorType: MARRESEARCH / Detector: CCD
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.813 Å / Relative weight: 1
ReflectionResolution: 2.26→91 Å / Num. obs: 136331 / % possible obs: 92.8 % / Observed criterion σ(I): -3 / Redundancy: 4.3 % / Rmerge(I) obs: 0.04 / Net I/σ(I): 23.7
Reflection shellResolution: 2.26→2.36 Å / Rmerge(I) obs: 0.13 / Mean I/σ(I) obs: 8.8 / % possible all: 77.7

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Processing

Software
NameVersionClassification
REFMAC5.2.0005refinement
XDSdata reduction
XSCALEdata scaling
PHASERphasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: PDB ENTRY 2J3V
Resolution: 2.3→40 Å / Cor.coef. Fo:Fc: 0.939 / Cor.coef. Fo:Fc free: 0.9 / SU B: 16.126 / SU ML: 0.194 / Cross valid method: THROUGHOUT / ESU R: 0.325 / ESU R Free: 0.243 / Stereochemistry target values: MAXIMUM LIKELIHOOD / Details: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS.
RfactorNum. reflection% reflectionSelection details
Rfree0.258 2650 2 %RANDOM
Rwork0.2 ---
obs0.201 129810 100 %-
Solvent computationIon probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.2 Å / Solvent model: BABINET MODEL WITH MASK
Displacement parametersBiso mean: 38.4 Å2
Baniso -1Baniso -2Baniso -3
1-3.73 Å20 Å20 Å2
2---0.51 Å20 Å2
3----3.22 Å2
Refinement stepCycle: LAST / Resolution: 2.3→40 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms20631 0 279 592 21502
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.0150.02221214
X-RAY DIFFRACTIONr_bond_other_d
X-RAY DIFFRACTIONr_angle_refined_deg1.6031.97728623
X-RAY DIFFRACTIONr_angle_other_deg
X-RAY DIFFRACTIONr_dihedral_angle_1_deg6.34352547
X-RAY DIFFRACTIONr_dihedral_angle_2_deg37.81825.3671036
X-RAY DIFFRACTIONr_dihedral_angle_3_deg17.475153968
X-RAY DIFFRACTIONr_dihedral_angle_4_deg20.42915108
X-RAY DIFFRACTIONr_chiral_restr0.1020.23179
X-RAY DIFFRACTIONr_gen_planes_refined0.0040.0215682
X-RAY DIFFRACTIONr_gen_planes_other
X-RAY DIFFRACTIONr_nbd_refined0.2060.29630
X-RAY DIFFRACTIONr_nbd_other
X-RAY DIFFRACTIONr_nbtor_refined0.3060.214431
X-RAY DIFFRACTIONr_nbtor_other
X-RAY DIFFRACTIONr_xyhbond_nbd_refined0.1490.21142
X-RAY DIFFRACTIONr_xyhbond_nbd_other
X-RAY DIFFRACTIONr_metal_ion_refined
X-RAY DIFFRACTIONr_metal_ion_other
X-RAY DIFFRACTIONr_symmetry_vdw_refined0.2580.2125
X-RAY DIFFRACTIONr_symmetry_vdw_other
X-RAY DIFFRACTIONr_symmetry_hbond_refined0.2080.220
X-RAY DIFFRACTIONr_symmetry_hbond_other
X-RAY DIFFRACTIONr_symmetry_metal_ion_refined
X-RAY DIFFRACTIONr_symmetry_metal_ion_other
X-RAY DIFFRACTIONr_mcbond_it0.5421.513170
X-RAY DIFFRACTIONr_mcbond_other
X-RAY DIFFRACTIONr_mcangle_it0.899220696
X-RAY DIFFRACTIONr_mcangle_other
X-RAY DIFFRACTIONr_scbond_it1.41239110
X-RAY DIFFRACTIONr_scbond_other
X-RAY DIFFRACTIONr_scangle_it2.1944.57927
X-RAY DIFFRACTIONr_scangle_other
X-RAY DIFFRACTIONr_long_range_B_refined
X-RAY DIFFRACTIONr_long_range_B_other
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

Ens-IDDom-IDAuth asym-IDNumberTypeRms dev position (Å)Weight position
11A1727medium positional0.380.5
12B1727medium positional0.330.5
13C1727medium positional0.370.5
21A1600medium positional0.310.5
22B1600medium positional0.360.5
23C1600medium positional0.320.5
31D1727medium positional0.350.5
32E1727medium positional0.350.5
33F1727medium positional0.430.5
41D1600medium positional0.310.5
42E1600medium positional0.330.5
43F1600medium positional0.340.5
11A1727medium thermal0.552
12B1727medium thermal0.582
13C1727medium thermal0.552
21A1600medium thermal0.472
22B1600medium thermal0.462
23C1600medium thermal0.452
31D1727medium thermal0.492
32E1727medium thermal0.492
33F1727medium thermal0.492
41D1600medium thermal0.492
42E1600medium thermal0.522
43F1600medium thermal0.52
LS refinement shellResolution: 2.3→2.36 Å / Total num. of bins used: 20 /
RfactorNum. reflection
Rfree0.344 197
Rwork0.24 9652
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)
11.28280.3253-0.13811.45730.38021.23550.0051-0.0977-0.18970.07470.0274-0.13160.26850.1305-0.0325-0.22090.0247-0.0339-0.12760.0188-0.0758-17.486-21.87-46.384
21.5223-0.7605-0.23643.27240.10521.5583-0.0052-0.11640.15160.0199-0.04050.0398-0.1618-0.01090.0457-0.2669-0.0292-0.0037-0.1319-0.0499-0.1007-26.476.572-40.38
31.5601-0.1467-0.19721.0996-0.09752.7033-0.0333-0.03080.16980.11940.057-0.0089-0.20520.0246-0.0237-0.2731-0.0308-0.0051-0.10510.007-0.0891-17.24466.477-26.426
41.38660.64150.98322.22080.7883.5323-0.03060.1167-0.0079-0.19410.05750.03990.0549-0.1156-0.027-0.31140.00460.013-0.05180.0338-0.1091-26.46756.836-53.843
51.0314-0.17610.2961.8444-0.17811.28380.02860.1444-0.0042-0.247-0.0609-0.1394-0.02550.10650.0323-0.28940.01020.0373-0.1236-0.0022-0.0911-17.56839.561-112.558
61.42680.7549-0.95032.0118-0.92742.8996-0.0142-0.089-0.12660.0324-0.0121-0.04630.0840.05770.0263-0.26320.0196-0.021-0.17230.0229-0.0525-26.14920.187-90.879
71.05280.68360.28032.148-0.52742.695-0.06340.00910.053-0.01980.17540.1855-0.4847-0.6189-0.112-0.24130.11380.01550.00980.0232-0.1212-33.76673.463-101.97
81.619-0.54760.72220.9775-0.74812.8303-0.0227-0.1401-0.0160.18840.02320.0854-0.2195-0.1939-0.0005-0.2189-0.00840.0114-0.1048-0.0047-0.1228-22.34860.168-77.513
91.0471-0.3617-0.38052.24470.13261.9635-0.00720.069-0.1671-0.27340.04630.22980.40620.0706-0.0391-0.10160.0084-0.095-0.1388-0.0178-0.0532-33.433-29.916-80.948
101.78221.17650.12972.3483-0.05430.6143-0.03540.12310.1105-0.15480.03540.07830.03870.0171-0.0001-0.20390.0296-0.0074-0.1150.0288-0.0822-22.136-2.11-81.811
111.1735-0.46410.29681.67270.79133.602-0.19060.02670.00420.3093-0.13020.21030.3063-0.21870.3208-0.2029-0.04630.0796-0.1349-0.0406-0.071-33.1540.857-2.092
122.1405-1.0258-1.68012.40852.31342.7656-0.1590.1048-0.160.3906-0.01780.23710.5088-0.13670.1768-0.1329-0.03970.0184-0.1445-0.0076-0.0825-21.94925.599-25.488
Refinement TLS group
IDRefine-IDRefine TLS-IDAuth asym-IDAuth seq-ID
1X-RAY DIFFRACTION1A1 - 221
2X-RAY DIFFRACTION2A222 - 432
3X-RAY DIFFRACTION3B1 - 221
4X-RAY DIFFRACTION4B222 - 432
5X-RAY DIFFRACTION5C1 - 221
6X-RAY DIFFRACTION6C222 - 432
7X-RAY DIFFRACTION7D1 - 221
8X-RAY DIFFRACTION8D222 - 432
9X-RAY DIFFRACTION9E1 - 221
10X-RAY DIFFRACTION10E222 - 432
11X-RAY DIFFRACTION11F1 - 221
12X-RAY DIFFRACTION12F222 - 432

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