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- PDB-3srd: Human M2 pyruvate kinase in complex with fructose 1-6 bisphosphat... -

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Basic information

Entry
Database: PDB / ID: 3srd
TitleHuman M2 pyruvate kinase in complex with fructose 1-6 bisphosphate and Oxalate.
ComponentsPyruvate kinase isozymes M1/M2
KeywordsTRANSFERASE / TIM Barrel / Phosphoryl transfer / PEP Binding / cytosol
Function / homology
Function and homology information


programmed cell death / pyruvate kinase / pyruvate kinase activity / positive regulation of cytoplasmic translation / histone H3T11 kinase activity / canonical glycolysis / Glycolysis / positive regulation of sprouting angiogenesis / potassium ion binding / rough endoplasmic reticulum ...programmed cell death / pyruvate kinase / pyruvate kinase activity / positive regulation of cytoplasmic translation / histone H3T11 kinase activity / canonical glycolysis / Glycolysis / positive regulation of sprouting angiogenesis / potassium ion binding / rough endoplasmic reticulum / glycolytic process / non-specific protein-tyrosine kinase / cilium / cellular response to insulin stimulus / extracellular vesicle / MHC class II protein complex binding / collagen-containing extracellular matrix / protein tyrosine kinase activity / secretory granule lumen / vesicle / ficolin-1-rich granule lumen / transcription coactivator activity / non-specific serine/threonine protein kinase / cadherin binding / phosphorylation / mRNA binding / Neutrophil degranulation / magnesium ion binding / protein homodimerization activity / positive regulation of transcription by RNA polymerase II / mitochondrion / RNA binding / extracellular exosome / extracellular region / ATP binding / nucleus / cytosol / cytoplasm
Similarity search - Function
PK beta-barrel domain-like / M1 Pyruvate Kinase; Domain 3 / Pyruvate kinase, C-terminal domain / Pyruvate Kinase; Chain: A, domain 1 / Pyruvate kinase, active site / Pyruvate kinase active site signature. / Pyruvate kinase / Pyruvate kinase, barrel / Pyruvate kinase, insert domain superfamily / Pyruvate kinase, barrel domain ...PK beta-barrel domain-like / M1 Pyruvate Kinase; Domain 3 / Pyruvate kinase, C-terminal domain / Pyruvate Kinase; Chain: A, domain 1 / Pyruvate kinase, active site / Pyruvate kinase active site signature. / Pyruvate kinase / Pyruvate kinase, barrel / Pyruvate kinase, insert domain superfamily / Pyruvate kinase, barrel domain / Pyruvate kinase, C-terminal / Pyruvate kinase, C-terminal domain superfamily / Pyruvate kinase, alpha/beta domain / Pyruvate kinase-like, insert domain superfamily / Phosphoenolpyruvate-binding domains / Pyruvate kinase-like domain superfamily / Pyruvate/Phosphoenolpyruvate kinase-like domain superfamily / TIM Barrel / Alpha-Beta Barrel / Beta Barrel / 3-Layer(aba) Sandwich / Mainly Beta / Alpha Beta
Similarity search - Domain/homology
1,6-di-O-phosphono-beta-D-fructofuranose / : / OXALATE ION / Pyruvate kinase PKM
Similarity search - Component
Biological speciesHomo sapiens (human)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 2.902 Å
AuthorsMorgan, H.P. / O'Reilly, F. / Palmer, R. / McNae, I.W. / Nowicki, M.W. / Wear, M.A. / Fothergill-Gilmore, L.A. / Walkinshaw, M.D.
CitationJournal: To be Published
Title: Allosetric regulation of M2 pyruvate kinase.
Authors: Morgan, H.P. / O'Reilly, F. / Palmer, R. / McNae, I.W. / Nowicki, M.W. / Wear, M.A. / Fothergill-Gilmore, L.A. / Walkinshaw, M.D.
History
DepositionJul 7, 2011Deposition site: RCSB / Processing site: RCSB
Revision 1.0Aug 8, 2012Provider: repository / Type: Initial release
Revision 1.1Jul 29, 2020Group: Data collection / Database references ...Data collection / Database references / Derived calculations / Structure summary
Category: chem_comp / entity ...chem_comp / entity / pdbx_chem_comp_identifier / pdbx_entity_nonpoly / pdbx_struct_conn_angle / struct_conn / struct_ref_seq_dif / struct_site / struct_site_gen
Item: _chem_comp.mon_nstd_flag / _chem_comp.name ..._chem_comp.mon_nstd_flag / _chem_comp.name / _chem_comp.type / _entity.pdbx_description / _pdbx_entity_nonpoly.name / _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.ptnr2_auth_asym_id / _pdbx_struct_conn_angle.ptnr2_auth_comp_id / _pdbx_struct_conn_angle.ptnr2_auth_seq_id / _pdbx_struct_conn_angle.ptnr2_label_asym_id / _pdbx_struct_conn_angle.ptnr2_label_atom_id / _pdbx_struct_conn_angle.ptnr2_label_comp_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_ref_seq_dif.details
Description: Carbohydrate remediation / Provider: repository / Type: Remediation
Revision 1.2Feb 28, 2024Group: Data collection / Database references / Structure summary
Category: chem_comp / chem_comp_atom ...chem_comp / chem_comp_atom / chem_comp_bond / database_2
Item: _chem_comp.pdbx_synonyms / _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: Pyruvate kinase isozymes M1/M2
B: Pyruvate kinase isozymes M1/M2
C: Pyruvate kinase isozymes M1/M2
D: Pyruvate kinase isozymes M1/M2
hetero molecules


Theoretical massNumber of molelcules
Total (without water)243,14124
Polymers240,7534
Non-polymers2,38820
Water2,270126
1


  • Idetical with deposited unit
  • defined by author&software
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area23670 Å2
ΔGint-159 kcal/mol
Surface area71710 Å2
MethodPISA
Unit cell
Length a, b, c (Å)95.050, 117.370, 110.460
Angle α, β, γ (deg.)90.00, 113.56, 90.00
Int Tables number4
Space group name H-MP1211
Noncrystallographic symmetry (NCS)NCS domain:
IDEns-IDDetails
11A
21B
31C
41D

NCS domain segments:
Dom-IDComponent-IDEns-IDRefine codeAuth asym-IDAuth seq-ID
1111A13 - 531
2111B13 - 531
3111C13 - 531
4111D13 - 531

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Components

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Protein / Sugars , 2 types, 8 molecules ABCD

#1: Protein
Pyruvate kinase isozymes M1/M2 / Cytosolic thyroid hormone-binding protein / CTHBP / Opa-interacting protein 3 / OIP-3 / Pyruvate ...Cytosolic thyroid hormone-binding protein / CTHBP / Opa-interacting protein 3 / OIP-3 / Pyruvate kinase 2/3 / Pyruvate kinase muscle isozyme / Thyroid hormone-binding protein 1 / THBP1 / Tumor M2-PK / p58


Mass: 60188.250 Da / Num. of mol.: 4
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Homo sapiens (human) / Gene: OIP3, PK2, PK3, PKM, PKM2 / Plasmid: pET28a_M2 / Production host: Escherichia coli (E. coli) / Strain (production host): BL21(DE3) / References: UniProt: P14618, pyruvate kinase
#3: Sugar
ChemComp-FBP / 1,6-di-O-phosphono-beta-D-fructofuranose / BETA-FRUCTOSE-1,6-DIPHOSPHATE / FRUCTOSE-1,6-BISPHOSPHATE / 1,6-di-O-phosphono-beta-D-fructose / 1,6-di-O-phosphono-D-fructose / 1,6-di-O-phosphono-fructose / Fructose 1,6-bisphosphate


Type: D-saccharide, beta linking / Mass: 340.116 Da / Num. of mol.: 4
Source method: isolated from a genetically manipulated source
Formula: C6H14O12P2
IdentifierTypeProgram
b-D-Fruf1PO36PO3IUPAC CARBOHYDRATE SYMBOLPDB-CARE 1.0

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Non-polymers , 5 types, 142 molecules

#2: Chemical
ChemComp-OXL / OXALATE ION / Oxalate


Mass: 88.019 Da / Num. of mol.: 4 / Source method: obtained synthetically / Formula: C2O4
#4: Chemical ChemComp-K / POTASSIUM ION


Mass: 39.098 Da / Num. of mol.: 3 / Source method: obtained synthetically / Formula: K
#5: Chemical
ChemComp-MG / MAGNESIUM ION


Mass: 24.305 Da / Num. of mol.: 4 / Source method: obtained synthetically / Formula: Mg
#6: Chemical
ChemComp-GOL / GLYCEROL / GLYCERIN / PROPANE-1,2,3-TRIOL / Glycerol


Mass: 92.094 Da / Num. of mol.: 5 / Source method: obtained synthetically / Formula: C3H8O3
#7: Water ChemComp-HOH / water / Water


Mass: 18.015 Da / Num. of mol.: 126 / 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: 2.35 Å3/Da / Density % sol: 47.57 %
Crystal growTemperature: 290 K / Method: vapor diffusion, hanging drop / pH: 7.2
Details: 10-16% polyethyleneglycol 8000, 20 mM triethanolamine-HCl buffer (pH 7.2), 50 mM MgCl2, 100 mM KCl, 10% glycerol, VAPOR DIFFUSION, HANGING DROP, temperature 290K

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: Diamond / Beamline: I03 / Wavelength: 1 Å
DetectorType: PSI PILATUS 6M / Detector: PIXEL / Date: Feb 15, 2011
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 1 Å / Relative weight: 1
ReflectionResolution: 2.9→58.7 Å / Num. all: 129300 / Num. obs: 45352 / % possible obs: 92 % / Observed criterion σ(F): 2.9 / Observed criterion σ(I): 2.9 / Redundancy: 2.9 % / Rmerge(I) obs: 0.062 / Rsym value: 0.088 / Net I/σ(I): 11.8
Reflection shell

Diffraction-ID: 1

Resolution (Å)Redundancy (%)Rmerge(I) obsMean I/σ(I) obsNum. unique allRsym value% possible all
2.9-2.982.30.54931.831180.54990
12.98-58.722.80.0151.45490.01593

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Processing

Software
NameVersionClassification
DNAdata collection
PHASERphasing
REFMAC5.5.0072refinement
MOSFLMdata reduction
SCALAdata scaling
RefinementMethod to determine structure: MOLECULAR REPLACEMENT / Resolution: 2.902→54.18 Å / Cor.coef. Fo:Fc: 0.903 / Cor.coef. Fo:Fc free: 0.882 / SU B: 55.204 / SU ML: 0.468 / Cross valid method: THROUGHOUT / ESU R Free: 0.503 / Stereochemistry target values: MAXIMUM LIKELIHOOD / Details: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS
RfactorNum. reflection% reflectionSelection details
Rfree0.27899 2283 5 %RANDOM
Rwork0.25298 ---
obs0.25431 43039 100 %-
all-45322 --
Solvent computationIon probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.4 Å / Solvent model: BABINET MODEL WITH MASK
Displacement parametersBiso mean: 78.245 Å2
Baniso -1Baniso -2Baniso -3
1-0.59 Å20 Å20.6 Å2
2---0.05 Å20 Å2
3----0.06 Å2
Refinement stepCycle: LAST / Resolution: 2.902→54.18 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms15772 0 141 126 16039
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.0070.02216173
X-RAY DIFFRACTIONr_bond_other_d
X-RAY DIFFRACTIONr_angle_refined_deg1.0461.9821852
X-RAY DIFFRACTIONr_angle_other_deg
X-RAY DIFFRACTIONr_dihedral_angle_1_deg4.49852054
X-RAY DIFFRACTIONr_dihedral_angle_2_deg40.68524.012668
X-RAY DIFFRACTIONr_dihedral_angle_3_deg16.87152919
X-RAY DIFFRACTIONr_dihedral_angle_4_deg18.32415124
X-RAY DIFFRACTIONr_chiral_restr0.060.22512
X-RAY DIFFRACTIONr_gen_planes_refined0.0040.02111908
X-RAY DIFFRACTIONr_gen_planes_other
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_it0.2761.510238
X-RAY DIFFRACTIONr_mcbond_other
X-RAY DIFFRACTIONr_mcangle_it0.525216522
X-RAY DIFFRACTIONr_scbond_it0.57835935
X-RAY DIFFRACTIONr_scangle_it1.0714.55330
X-RAY DIFFRACTIONr_rigid_bond_restr
X-RAY DIFFRACTIONr_sphericity_free
X-RAY DIFFRACTIONr_sphericity_bonded
Refine LS restraints NCS

Ens-ID: 1 / Number: 3920 / Refine-ID: X-RAY DIFFRACTION

Dom-IDAuth asym-IDTypeRms dev position (Å)Weight position
1ATIGHT POSITIONAL0.030.05
2BTIGHT POSITIONAL0.020.05
3CTIGHT POSITIONAL0.030.05
4DTIGHT POSITIONAL0.040.05
1ATIGHT THERMAL0.030.5
2BTIGHT THERMAL0.030.5
3CTIGHT THERMAL0.030.5
4DTIGHT THERMAL0.030.5
LS refinement shellResolution: 2.902→2.977 Å / Total num. of bins used: 20
RfactorNum. reflection% reflection
Rfree0.414 159 -
Rwork0.376 2949 -
obs--100 %
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)
11.05621.4513-1.27652.898-1.80991.5480.49710.17160.3590.0159-0.08940.7751-0.5027-0.2064-0.40770.70050.1591-0.0270.5010.00560.4547-14.44599.4012-15.0272
21.04490.2202-0.39551.0503-0.09811.4635-0.01660.03380.14190.1340.0158-0.0233-0.1679-0.07140.00080.15260.00830.06850.1336-0.03790.1517-30.3724-7.25439.4502
30.5408-0.2287-0.10450.4684-0.3881.1335-0.0155-0.0677-0.12970.05330.02120.03820.23190.073-0.00570.18930.01640.05080.1127-0.01520.1397-21.3283-21.51867.8595
42.28980.34560.70581.5567-0.26732.1320.04490.1856-0.0234-0.10510.01920.0722-0.0439-0.1153-0.06410.18260.03530.08230.17050.02760.1787-36.7453-0.4318-14.9519
51.03690.9229-2.42852.1617-1.87545.7606-0.03470.51630.23680.20170.25270.97160.7224-1.2974-0.2180.5418-0.11550.04830.6675-0.03220.6215-19.228-14.5665-37.699
60.6267-0.05430.12570.679-0.08690.7205-0.00250.38060.3002-0.2918-0.0001-0.1598-0.25010.0050.00260.40850.08270.10060.40070.07510.3805-14.927614.9333-65.8023
70.9361-0.2403-0.01640.3579-0.21710.51060.05510.0160.34860.0280.00920.0382-0.2922-0.2224-0.06430.30270.08170.05870.27980.03020.289-17.95311.4912-53.9819
81.1082-0.6224-0.95611.85590.0822.12370.11470.2391-0.0005-0.0901-0.0310.0270.0355-0.0914-0.08370.23180.05790.07550.30490.06260.2539-36.05812.7628-38.188
91.05650.18210.21940.03930.03020.11710.03-0.2177-0.8253-0.06660.0939-0.15960.19350.1024-0.12390.760.2110.01330.6975-0.09140.7516-6.1238-18.2214-15.8691
100.8217-0.1603-0.1840.76690.15260.5159-0.041-0.1740.06420.1960.0022-0.0257-0.05050.14340.03880.2999-0.02380.04180.2720.00490.21232.9147.12213.3579
111.41610.014-0.28630.98660.35650.5584-0.0008-0.1452-0.05050.1068-0.0242-0.0304-0.03560.18190.0250.1792-0.00110.04280.1918-0.01630.1585-2.4454-1.62323.3964
121.6620.6653-0.79132.04090.1681.81770.0154-0.0651-0.0173-0.0583-0.0118-0.12940.06910.1139-0.00360.18540.0130.06470.23190.02380.192515.9726-9.1146-13.5936
130.28640.482-0.4280.939-0.74941.88680.096-0.0532-0.00650.066-0.0629-0.2948-0.47360.5871-0.03320.56980.00370.05030.57-0.02020.6801-1.12088.2096-35.1444
141.12880.171-0.01060.70120.08711.5742-0.04580.12460.0895-0.1093-0.00460.17350.0742-0.25020.05040.17510.0250.09320.09580.05080.1511-1.4366-15.7336-68.4766
151.5138-0.1302-0.2290.61070.15041.4480.00250.0233-0.0828-0.04080.00430.06880.1397-0.059-0.00680.1441-0.01310.06310.10780.0140.13171.0161-15.1516-55.8249
161.81440.0190.53811.4279-0.03612.33530.0002-0.17240.0140.15220.02320.027-0.030.0253-0.02340.1456-0.01090.08090.1260.0220.161317.5169-8.8926-37.0702
Refinement TLS group
IDRefine-IDRefine TLS-IDAuth asym-IDAuth seq-ID
1X-RAY DIFFRACTION1A13 - 26
2X-RAY DIFFRACTION2A27 - 134
3X-RAY DIFFRACTION3A135 - 382
4X-RAY DIFFRACTION4A383 - 531
5X-RAY DIFFRACTION5B14 - 25
6X-RAY DIFFRACTION6B26 - 225
7X-RAY DIFFRACTION7B226 - 387
8X-RAY DIFFRACTION8B388 - 531
9X-RAY DIFFRACTION9C13 - 26
10X-RAY DIFFRACTION10C27 - 279
11X-RAY DIFFRACTION11C280 - 381
12X-RAY DIFFRACTION12C382 - 531
13X-RAY DIFFRACTION13D15 - 27
14X-RAY DIFFRACTION14D28 - 219
15X-RAY DIFFRACTION15D220 - 388
16X-RAY DIFFRACTION16D389 - 531

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