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- PDB-5o8q: Crystal structure of R. ruber ADH-A, mutant Y294F, W295A -

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Basic information

Entry
Database: PDB / ID: 5o8q
TitleCrystal structure of R. ruber ADH-A, mutant Y294F, W295A
ComponentsAlcohol dehydrogenase
KeywordsOXIDOREDUCTASE / alcohol dehydrogenase mutant variant / NADH-dependent / Zn2+-dependent / Rossmann fold
Function / homology
Function and homology information


oxidoreductase activity / zinc ion binding
Similarity search - Function
Alcohol dehydrogenase, zinc-type, conserved site / Zinc-containing alcohol dehydrogenases signature. / Quinone Oxidoreductase; Chain A, domain 1 / Medium-chain alcohol dehydrogenases, catalytic domain / Alcohol dehydrogenase-like, C-terminal / Zinc-binding dehydrogenase / Alcohol dehydrogenase, N-terminal / Alcohol dehydrogenase GroES-like domain / Polyketide synthase, enoylreductase domain / Enoylreductase ...Alcohol dehydrogenase, zinc-type, conserved site / Zinc-containing alcohol dehydrogenases signature. / Quinone Oxidoreductase; Chain A, domain 1 / Medium-chain alcohol dehydrogenases, catalytic domain / Alcohol dehydrogenase-like, C-terminal / Zinc-binding dehydrogenase / Alcohol dehydrogenase, N-terminal / Alcohol dehydrogenase GroES-like domain / Polyketide synthase, enoylreductase domain / Enoylreductase / GroES-like superfamily / NAD(P)-binding Rossmann-like Domain / NAD(P)-binding domain superfamily / Alpha-Beta Complex / Rossmann fold / 3-Layer(aba) Sandwich / Alpha Beta
Similarity search - Domain/homology
NICOTINAMIDE-ADENINE-DINUCLEOTIDE / Alcohol dehydrogenase
Similarity search - Component
Biological speciesRhodococcus sp. M8 (bacteria)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 2.22 Å
AuthorsDobritzsch, D. / Maurer, D. / Hamnevik, E. / Enugala, T.R. / Widersten, M.
Funding support Sweden, 1items
OrganizationGrant numberCountry
Swedish Research Council Sweden
CitationJournal: FEBS J. / Year: 2017
Title: Relaxation of nonproductive binding and increased rate of coenzyme release in an alcohol dehydrogenase increases turnover with a nonpreferred alcohol enantiomer.
Authors: Hamnevik, E. / Enugala, T.R. / Maurer, D. / Ntuku, S. / Oliveira, A. / Dobritzsch, D. / Widersten, M.
History
DepositionJun 14, 2017Deposition site: PDBE / Processing site: PDBE
Revision 1.0Oct 11, 2017Provider: repository / Type: Initial release
Revision 1.1Nov 29, 2017Group: Database references / Category: citation
Item: _citation.journal_volume / _citation.page_first ..._citation.journal_volume / _citation.page_first / _citation.page_last / _citation.title
Revision 1.2Jan 17, 2024Group: Data collection / Database references ...Data collection / Database references / Derived calculations / Refinement description
Category: chem_comp_atom / chem_comp_bond ...chem_comp_atom / chem_comp_bond / database_2 / pdbx_initial_refinement_model / 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 / _struct_conn.pdbx_dist_value / _struct_conn.pdbx_ptnr1_label_alt_id / _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_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: Alcohol dehydrogenase
B: Alcohol dehydrogenase
C: Alcohol dehydrogenase
D: Alcohol dehydrogenase
E: Alcohol dehydrogenase
F: Alcohol dehydrogenase
G: Alcohol dehydrogenase
H: Alcohol dehydrogenase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)294,10532
Polymers287,7518
Non-polymers6,35424
Water13,547752
1
A: Alcohol dehydrogenase
D: Alcohol dehydrogenase
E: Alcohol dehydrogenase
G: Alcohol dehydrogenase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)147,05316
Polymers143,8764
Non-polymers3,17712
Water724
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area15350 Å2
ΔGint-263 kcal/mol
Surface area46140 Å2
MethodPISA
2
B: Alcohol dehydrogenase
C: Alcohol dehydrogenase
F: Alcohol dehydrogenase
H: Alcohol dehydrogenase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)147,05316
Polymers143,8764
Non-polymers3,17712
Water724
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area15390 Å2
ΔGint-264 kcal/mol
Surface area46150 Å2
MethodPISA
Unit cell
Length a, b, c (Å)103.689, 109.458, 256.653
Angle α, β, γ (deg.)90.00, 90.00, 90.00
Int Tables number19
Space group name H-MP212121
Noncrystallographic symmetry (NCS)NCS domain:
IDEns-IDDetails
11A
21B
12A
22C
13A
23D
14A
24E
15A
25F
16A
26G
17A
27H
18B
28C
19B
29D
110B
210E
111B
211F
112B
212G
113B
213H
114C
214D
115C
215E
116C
216F
117C
217G
118C
218H
119D
219E
120D
220F
121D
221G
122D
222H
123E
223F
124E
224G
125E
225H
126F
226G
127F
227H
128G
228H

NCS domain segments:

Component-ID: 0 / Beg auth comp-ID: MET / Beg label comp-ID: MET / End auth comp-ID: PRO / End label comp-ID: PRO / Refine code: 0 / Auth seq-ID: 1 - 345 / Label seq-ID: 1 - 345

Dom-IDEns-IDAuth asym-IDLabel asym-ID
11AA
21BB
12AA
22CC
13AA
23DD
14AA
24EE
15AA
25FF
16AA
26GG
17AA
27HH
18BB
28CC
19BB
29DD
110BB
210EE
111BB
211FF
112BB
212GG
113BB
213HH
114CC
214DD
115CC
215EE
116CC
216FF
117CC
217GG
118CC
218HH
119DD
219EE
120DD
220FF
121DD
221GG
122DD
222HH
123EE
223FF
124EE
224GG
125EE
225HH
126FF
226GG
127FF
227HH
128GG
228HH

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

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Components

#1: Protein
Alcohol dehydrogenase /


Mass: 35968.887 Da / Num. of mol.: 8 / Mutation: Y294F, W295A
Source method: isolated from a genetically manipulated source
Details: contains C-terminal His-tag / Source: (gene. exp.) Rhodococcus sp. M8 (bacteria) / Gene: BKE56_025765 / Plasmid: pGT7ADHA-5H / Production host: Escherichia coli BL21 (bacteria) / Variant (production host): AI / References: UniProt: A0A1Q8I6M1
#2: Chemical
ChemComp-ZN / ZINC ION


Mass: 65.409 Da / Num. of mol.: 16 / Source method: obtained synthetically / Formula: Zn
#3: Chemical
ChemComp-NAD / NICOTINAMIDE-ADENINE-DINUCLEOTIDE / Nicotinamide adenine dinucleotide


Mass: 663.425 Da / Num. of mol.: 8 / Source method: obtained synthetically / Formula: C21H27N7O14P2 / Comment: NAD*YM
#4: Water ChemComp-HOH / water / Water


Mass: 18.015 Da / Num. of mol.: 752 / 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: 2.49 Å3/Da / Density % sol: 50.3 %
Crystal growTemperature: 293 K / Method: vapor diffusion, hanging drop / pH: 8
Details: 16% PAA, 20mM MgCl2, 0.1 m Tris pH8, 4mM NAD+, 10mM Acetophenone, 7 mg/ml ADH-A

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: Diamond / Beamline: I03 / Wavelength: 0.9763 Å
DetectorType: DECTRIS PILATUS3 S 6M / Detector: PIXEL / Date: Dec 18, 2016
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.9763 Å / Relative weight: 1
ReflectionResolution: 2.21→29.74 Å / Num. obs: 144348 / % possible obs: 99 % / Redundancy: 7.4 % / Biso Wilson estimate: 25.9 Å2 / CC1/2: 0.995 / Rmerge(I) obs: 0.168 / Rpim(I) all: 0.092 / Net I/σ(I): 7.8
Reflection shellResolution: 2.21→2.25 Å / Redundancy: 5.9 % / Rmerge(I) obs: 0.68 / Mean I/σ(I) obs: 2.4 / Num. unique obs: 5774 / CC1/2: 0.763 / Rpim(I) all: 0.285 / % possible all: 80.9

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Processing

Software
NameVersionClassification
REFMAC5.8.0158refinement
xia2data reduction
Aimlessdata scaling
PHASERphasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: 3jv7
Resolution: 2.22→29.64 Å / Cor.coef. Fo:Fc: 0.949 / Cor.coef. Fo:Fc free: 0.938 / SU B: 11.201 / SU ML: 0.148 / Cross valid method: THROUGHOUT / ESU R: 0.274 / ESU R Free: 0.191 / Details: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS
RfactorNum. reflection% reflectionSelection details
Rfree0.21576 7306 5.1 %RANDOM
Rwork0.19351 ---
obs0.19463 136802 99.26 %-
Solvent computationIon probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.2 Å
Displacement parametersBiso mean: 39.583 Å2
Baniso -1Baniso -2Baniso -3
1-2.13 Å20 Å20 Å2
2--0.39 Å2-0 Å2
3----2.52 Å2
Refinement stepCycle: 1 / Resolution: 2.22→29.64 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms19705 0 368 752 20825
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.010.01920525
X-RAY DIFFRACTIONr_bond_other_d0.0010.0219351
X-RAY DIFFRACTIONr_angle_refined_deg1.4261.9928073
X-RAY DIFFRACTIONr_angle_other_deg0.945344548
X-RAY DIFFRACTIONr_dihedral_angle_1_deg5.65952760
X-RAY DIFFRACTIONr_dihedral_angle_2_deg29.72322.769715
X-RAY DIFFRACTIONr_dihedral_angle_3_deg12.63152858
X-RAY DIFFRACTIONr_dihedral_angle_4_deg18.85715146
X-RAY DIFFRACTIONr_chiral_restr0.0730.23303
X-RAY DIFFRACTIONr_gen_planes_refined0.0050.02123346
X-RAY DIFFRACTIONr_gen_planes_other0.0010.024112
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_it0.641.96211049
X-RAY DIFFRACTIONr_mcbond_other0.641.96211048
X-RAY DIFFRACTIONr_mcangle_it1.132.94113800
X-RAY DIFFRACTIONr_mcangle_other1.132.94113801
X-RAY DIFFRACTIONr_scbond_it0.6822.0999476
X-RAY DIFFRACTIONr_scbond_other0.6822.0999477
X-RAY DIFFRACTIONr_scangle_it
X-RAY DIFFRACTIONr_scangle_other1.1643.10814272
X-RAY DIFFRACTIONr_long_range_B_refined3.41623.72420720
X-RAY DIFFRACTIONr_long_range_B_other3.37323.57420613
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)
11A195880.05
12B195880.05
21A195280.04
22C195280.04
31A195520.05
32D195520.05
41A195220.05
42E195220.05
51A196100.04
52F196100.04
61A195580.05
62G195580.05
71A193620.06
72H193620.06
81B195600.04
82C195600.04
91B196140.04
92D196140.04
101B194700.05
102E194700.05
111B195500.05
112F195500.05
121B195240.05
122G195240.05
131B194680.05
132H194680.05
141C195660.05
142D195660.05
151C196400.04
152E196400.04
161C196200.04
162F196200.04
171C196420.04
172G196420.04
181C194780.06
182H194780.06
191D195440.04
192E195440.04
201D196320.04
202F196320.04
211D196320.04
212G196320.04
221D195240.05
222H195240.05
231E195780.04
232F195780.04
241E196600.04
242G196600.04
251E194120.05
252H194120.05
261F196380.04
262G196380.04
271F194700.05
272H194700.05
281G194440.05
282H194440.05
LS refinement shellResolution: 2.217→2.274 Å / Total num. of bins used: 20
RfactorNum. reflection% reflection
Rfree0.274 536 -
Rwork0.266 9100 -
obs--91.18 %
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)
14.7837-1.40080.50283.6978-0.86053.11770.19780.2954-0.0703-0.2853-0.2388-0.30030.34590.49620.0410.2073-0.0513-0.03070.2053-0.03340.22796.8688182.675887.4373
21.73620.79090.28932.7984-0.70251.2133-0.0292-0.08580.01430.0576-0.0107-0.195-0.03870.19860.03990.0367-0.00720.01270.0679-0.03370.0894-8.6469170.111190.547
31.41630.53610.76463.63180.81281.38540.087-0.1512-0.09610.4289-0.0342-0.24860.09460.2093-0.05280.0759-0.0105-0.00240.10890.01920.0536-6.3887162.941491.8205
44.5439-0.4893-0.10912.4507-0.51622.15650.09170.1178-0.07-0.256-0.1374-0.3369-0.06050.46630.04570.2224-0.1089-0.07240.2403-0.02750.21915.579182.567828.4197
51.11850.50380.29312.4853-0.60241.903-0.0997-0.04690.07320.10120.0237-0.159-0.09880.27850.0760.01880.0023-0.02070.0617-0.01080.0792-10.1064169.672329.0522
60.86660.04580.33892.61670.69312.0134-0.0315-0.1218-0.03170.3205-0.0048-0.13210.09050.28360.03630.04240.012-0.00750.0940.03050.0578-8.0124162.561630.7748
73.4462-0.91681.03522.6282-0.44934.3536-0.0634-0.24130.3543-0.09690.02140.1282-1.3255-0.2080.0420.7136-0.1210.02360.1910.0450.2799-18.3863198.751-2.477
82.010.29340.63971.00460.10272.6326-0.05170.13840.176-0.1863-0.0099-0.0501-0.45940.0160.06150.12040.01410.00540.02360.0250.0694-26.0544179.8962-1.6113
93.11870.4836-1.10321.0898-0.54322.6571-0.00520.38110.1283-0.15680.05090.0726-0.3431-0.2789-0.04570.14710.0481-0.03120.06590.00330.0242-33.4365179.8135-2.972
104.0938-1.9247-0.55744.421.26592.7870.1310.23950.124-0.3693-0.26690.3267-0.2111-0.52040.13590.1217-0.02120.00110.18720.02540.173-56.728151.868296.0546
110.72640.2939-0.2782.46110.37441.1493-0.0133-0.0977-0.05260.0717-0.04290.1973-0.0671-0.22050.05620.01370.0259-0.0040.08340.00850.0896-41.2391164.602493.9007
120.95950.1932-0.5991.8822-0.63852.07880.0484-0.09050.08160.2444-0.04730.1581-0.272-0.1838-0.00120.06720.0351-0.01810.0734-0.04520.0941-43.4441171.84993.9932
133.3467-1.7532-1.01843.39270.75653.58140.0502-0.3332-0.12720.06950.163-0.1440.73170.417-0.21330.31850.0075-0.12010.1184-0.02830.229-32.0276129.284369.9594
141.88460.5498-0.43521.88250.07011.7585-0.00460.0708-0.1554-0.25240.0108-0.01530.21420.1229-0.00620.09780.0234-0.04280.0173-0.00530.0719-24.8351147.886166.4163
153.02821.13271.15852.01160.70542.02060.0360.4131-0.124-0.28620.0979-0.18220.17050.2905-0.13390.14240.04440.02060.0732-0.01690.0254-17.4383147.776465.1107
163.5403-0.8822-0.06483.36710.55523.06930.20640.36280.3004-0.3795-0.14280.2103-0.2585-0.5176-0.06360.1669-0.01550.06060.2240.0680.198-58.4516151.510326.4065
170.83250.5164-0.33152.80440.25051.2655-0.03630.0141-0.0210.1066-0.00360.1751-0.0017-0.23520.03990.01620.01140.00570.06240.01630.0582-42.7177164.247326.7907
181.01060.5062-0.49183.1916-1.11221.80460.0701-0.04180.120.3845-0.03030.2838-0.2212-0.2486-0.03980.06540.02780.01680.0732-0.0360.0533-44.8977171.448826.39
195.0266-2.30461.89194.44340.17613.5555-0.1234-0.53180.7397-0.4110.0672-0.1452-1.1531-0.19010.05630.8402-0.03890.09930.23130.07370.4222-22.1989199.467560.8206
202.73350.71920.8871.54490.69082.8086-0.00950.15640.2786-0.4559-0.079-0.0338-0.62390.04420.08850.27280.05190.0170.03270.0170.0839-28.2602181.199163.3477
212.56070.0574-0.95081.3739-0.02032.88960.06550.33140.0966-0.4183-0.07070.1552-0.512-0.36990.00520.32980.1256-0.0820.0913-0.01090.0567-38.169180.148462.8207
222.4701-1.422-1.35174.4583-0.87923.24890.1736-0.2611-0.4136-0.3074-0.236-0.25191.05130.47960.06240.8403-0.0024-0.07070.3515-0.06840.3776-29.679128.2815.1258
233.07440.6723-1.16551.4181-0.26372.7465-0.11980.12-0.3188-0.1985-0.00650.0150.62240.12650.12630.18510.0518-0.0140.0462-0.0120.0673-22.3202147.00512.634
243.81090.43071.04211.51950.53362.70770.0150.476-0.403-0.19820.1032-0.10330.44230.3402-0.11820.17950.06790.03710.1065-0.02160.077-14.4011146.99132.6937
Refinement TLS group
IDRefine-IDRefine TLS-IDAuth asym-IDAuth seq-ID
1X-RAY DIFFRACTION1A1 - 82
2X-RAY DIFFRACTION2A83 - 231
3X-RAY DIFFRACTION3A232 - 345
4X-RAY DIFFRACTION4B1 - 82
5X-RAY DIFFRACTION5B83 - 231
6X-RAY DIFFRACTION6B232 - 345
7X-RAY DIFFRACTION7C1 - 82
8X-RAY DIFFRACTION8C83 - 231
9X-RAY DIFFRACTION9C232 - 345
10X-RAY DIFFRACTION10D1 - 82
11X-RAY DIFFRACTION11D83 - 231
12X-RAY DIFFRACTION12D232 - 345
13X-RAY DIFFRACTION13E1 - 82
14X-RAY DIFFRACTION14E83 - 231
15X-RAY DIFFRACTION15E232 - 345
16X-RAY DIFFRACTION16F1 - 82
17X-RAY DIFFRACTION17F83 - 231
18X-RAY DIFFRACTION18F232 - 345
19X-RAY DIFFRACTION19G1 - 82
20X-RAY DIFFRACTION20G83 - 221
21X-RAY DIFFRACTION21G222 - 345
22X-RAY DIFFRACTION22H1 - 79
23X-RAY DIFFRACTION23H80 - 231
24X-RAY DIFFRACTION24H232 - 345

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Jul 12, 2017. Major update of PDB

Major update of PDB

  • wwPDB released updated PDB data conforming to the new PDBx/mmCIF dictionary.
  • This is a major update changing the version number from 4 to 5, and with Remediation, in which all the entries are updated.
  • 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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