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- PDB-8w0o: GDH-105 crystal structure -

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Basic information

Entry
Database: PDB / ID: 8w0o
TitleGDH-105 crystal structure
ComponentsDehydrogenase
KeywordsOXIDOREDUCTASE / protein engineering / glucose dehydrogenase / scalable manufacturing
Function / homologyglucose 1-dehydrogenase [NAD(P)+] activity / 3-oxoacyl-[acyl-carrier-protein] reductase (NADPH) activity / Short-chain dehydrogenase/reductase, conserved site / Short-chain dehydrogenases/reductases family signature. / Enoyl-(Acyl carrier protein) reductase / Short-chain dehydrogenase/reductase SDR / NAD(P)-binding domain superfamily / NICOTINAMIDE-ADENINE-DINUCLEOTIDE / Dehydrogenase
Function and homology information
Biological speciesBacillus subtilis (bacteria)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 1.66 Å
AuthorsPeat, T.S. / Newman, J.
Funding support1items
OrganizationGrant numberCountry
Not funded
CitationJournal: Acs Catalysis / Year: 2024
Title: Enhancing the Imine Reductase Activity of a Promiscuous Glucose Dehydrogenase for Scalable Manufacturing of a Chiral Neprilysin Inhibitor Precursor
Authors: Yi, X. / Kleinbeck, F. / Ching, C. / Boghospor, L. / Gomes, S. / Alvizo, O. / Allmendinger, T. / Fell, J. / Subramanian, N. / Li, M. / Garcia, R. / Riggins, J. / Entwistle, D. / Richter, Y. ...Authors: Yi, X. / Kleinbeck, F. / Ching, C. / Boghospor, L. / Gomes, S. / Alvizo, O. / Allmendinger, T. / Fell, J. / Subramanian, N. / Li, M. / Garcia, R. / Riggins, J. / Entwistle, D. / Richter, Y. / Gschwend, D. / Lauener, L. / Peat, T.S. / Lebhar, H. / Schlama, T. / Ruch, T.
History
DepositionFeb 13, 2024Deposition site: RCSB / Processing site: RCSB
Revision 1.0Jul 24, 2024Provider: repository / Type: Initial release

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: Dehydrogenase
B: Dehydrogenase
C: Dehydrogenase
D: Dehydrogenase
E: Dehydrogenase
H: Dehydrogenase
F: Dehydrogenase
G: Dehydrogenase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)230,45920
Polymers225,0108
Non-polymers5,44912
Water15,115839
1
A: Dehydrogenase
E: Dehydrogenase
H: Dehydrogenase
G: Dehydrogenase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)115,23010
Polymers112,5054
Non-polymers2,7256
Water724
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area19980 Å2
ΔGint-165 kcal/mol
Surface area33220 Å2
MethodPISA
2
B: Dehydrogenase
C: Dehydrogenase
D: Dehydrogenase
F: Dehydrogenase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)115,23010
Polymers112,5054
Non-polymers2,7256
Water724
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area20070 Å2
ΔGint-163 kcal/mol
Surface area33240 Å2
MethodPISA
Unit cell
Length a, b, c (Å)63.336, 120.428, 125.775
Angle α, β, γ (deg.)90.000, 101.891, 90.000
Int Tables number4
Space group name H-MP1211
Noncrystallographic symmetry (NCS)NCS domain:
IDEns-IDDetails
11A
21A
32A
42A
53A
63A
74A
84A
95A
105A
116A
126A
137A
147A
158A
168A
179A
189A
1910A
2010A
2111A
2211A
2312A
2412A
2513A
2613A
2714A
2814A
2915A
3015A
3116A
3216A
3317A
3417A
3518A
3618A
3719A
3819A
3920A
4020A
4121A
4221A
4322A
4422A
4523A
4623A
4724A
4824A
4925A
5025A
5126A
5226A
5327A
5427A
5528A
5628A

NCS domain segments:

Beg auth comp-ID: MET / Beg label comp-ID: MET / End auth comp-ID: GLY / End label comp-ID: GLY / Auth asym-ID: A / Label asym-ID: A / Auth seq-ID: 1 - 259 / Label seq-ID: 1 - 259

Dom-IDComponent-IDEns-ID
111
211
322
422
533
633
744
844
955
1055
1166
1266
1377
1477
1588
1688
1799
1899
191010
201010
211111
221111
231212
241212
251313
261313
271414
281414
291515
301515
311616
321616
331717
341717
351818
361818
371919
381919
392020
402020
412121
422121
432222
442222
452323
462323
472424
482424
492525
502525
512626
522626
532727
542727
552828
562828

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
16Local NCS retraints between domains: 31 32
17Local NCS retraints between domains: 33 34
18Local NCS retraints between domains: 35 36
19Local NCS retraints between domains: 37 38
20Local NCS retraints between domains: 39 40
21Local NCS retraints between domains: 41 42
22Local NCS retraints between domains: 43 44
23Local NCS retraints between domains: 45 46
24Local NCS retraints between domains: 47 48
25Local NCS retraints between domains: 49 50
26Local NCS retraints between domains: 51 52
27Local NCS retraints between domains: 53 54
28Local NCS retraints between domains: 55 56

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Components

#1: Protein
Dehydrogenase


Mass: 28126.260 Da / Num. of mol.: 8
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Bacillus subtilis (bacteria)
Gene: gdh, B4417_2813, CFD21_16405, NRS6116_02705, NRS6202_01215
Production host: Escherichia coli (E. coli)
References: UniProt: M9TFE3, glucose 1-dehydrogenase [NAD(P)+]
#2: Chemical
ChemComp-NAD / NICOTINAMIDE-ADENINE-DINUCLEOTIDE


Mass: 663.425 Da / Num. of mol.: 8 / Source method: obtained synthetically / Formula: C21H27N7O14P2 / Comment: NAD*YM
#3: Chemical
ChemComp-CL / CHLORIDE ION


Mass: 35.453 Da / Num. of mol.: 4 / Source method: obtained synthetically / Formula: Cl
#4: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 839 / Source method: isolated from a natural source / Formula: H2O
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: 2.09 Å3/Da / Density % sol: 41.04 %
Crystal growTemperature: 295 K / Method: vapor diffusion, sitting drop / pH: 6
Details: Crystals were grown in 30% PEG4000 with 200 mM malate imidazole buffer at pH 6.0. Sitting drops were 200 nL of reservoir with 200 nL of protein at about 10 mg/mL at room temperature.

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Data collection

DiffractionMean temperature: 100 K / Serial crystal experiment: N
Diffraction sourceSource: SYNCHROTRON / Site: Australian Synchrotron / Beamline: MX2 / Wavelength: 0.9537 Å
DetectorType: DECTRIS EIGER X 16M / Detector: PIXEL / Date: Mar 18, 2023
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.9537 Å / Relative weight: 1
ReflectionResolution: 1.66→49 Å / Num. obs: 212485 / % possible obs: 98 % / Redundancy: 6.9 % / CC1/2: 0.997 / Rmerge(I) obs: 0.141 / Rpim(I) all: 0.058 / Net I/σ(I): 9.3
Reflection shellResolution: 1.66→1.69 Å / Redundancy: 6.8 % / Rmerge(I) obs: 1.323 / Mean I/σ(I) obs: 1.4 / Num. unique obs: 9477 / CC1/2: 0.609 / Rpim(I) all: 0.541 / % possible all: 88.9

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Processing

Software
NameVersionClassification
REFMAC5.8.0405refinement
XDSdata reduction
Aimlessdata scaling
PHASERphasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT / Resolution: 1.66→49 Å / Cor.coef. Fo:Fc: 0.966 / Cor.coef. Fo:Fc free: 0.956 / SU B: 2.425 / SU ML: 0.077 / Cross valid method: FREE R-VALUE / ESU R: 0.106 / ESU R Free: 0.098
Details: Hydrogens have been added in their riding positions
RfactorNum. reflection% reflection
Rfree0.1965 10634 5.006 %
Rwork0.1731 201808 -
all0.174 --
obs-212442 97.883 %
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: 22.442 Å2
Baniso -1Baniso -2Baniso -3
1--0.843 Å20 Å21.475 Å2
2---0.868 Å2-0 Å2
3---1.001 Å2
Refinement stepCycle: LAST / Resolution: 1.66→49 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms15672 0 356 839 16867
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.0080.01216727
X-RAY DIFFRACTIONr_bond_other_d0.0010.01616069
X-RAY DIFFRACTIONr_angle_refined_deg1.3611.64222778
X-RAY DIFFRACTIONr_angle_other_deg0.461.56737237
X-RAY DIFFRACTIONr_dihedral_angle_1_deg5.99852146
X-RAY DIFFRACTIONr_dihedral_angle_2_deg5.495532
X-RAY DIFFRACTIONr_dihedral_angle_other_2_deg0.02358
X-RAY DIFFRACTIONr_dihedral_angle_3_deg14.002102878
X-RAY DIFFRACTIONr_dihedral_angle_6_deg16.26810635
X-RAY DIFFRACTIONr_chiral_restr0.0720.22574
X-RAY DIFFRACTIONr_gen_planes_refined0.0070.0219061
X-RAY DIFFRACTIONr_gen_planes_other0.0010.023367
X-RAY DIFFRACTIONr_nbd_refined0.2240.23405
X-RAY DIFFRACTIONr_symmetry_nbd_other0.190.214911
X-RAY DIFFRACTIONr_nbtor_refined0.1790.28511
X-RAY DIFFRACTIONr_symmetry_nbtor_other0.0750.28449
X-RAY DIFFRACTIONr_xyhbond_nbd_refined0.1250.2766
X-RAY DIFFRACTIONr_symmetry_xyhbond_nbd_other0.0190.21
X-RAY DIFFRACTIONr_symmetry_nbd_refined0.2910.249
X-RAY DIFFRACTIONr_nbd_other0.3090.2162
X-RAY DIFFRACTIONr_symmetry_xyhbond_nbd_refined0.3110.221
X-RAY DIFFRACTIONr_mcbond_it2.0482.1568485
X-RAY DIFFRACTIONr_mcbond_other2.0482.1558484
X-RAY DIFFRACTIONr_mcangle_it2.9083.86710664
X-RAY DIFFRACTIONr_mcangle_other2.9093.86810665
X-RAY DIFFRACTIONr_scbond_it3.0132.4558242
X-RAY DIFFRACTIONr_scbond_other3.0132.4558243
X-RAY DIFFRACTIONr_scangle_it4.584.35512114
X-RAY DIFFRACTIONr_scangle_other4.584.35512115
X-RAY DIFFRACTIONr_lrange_it5.77927.15973767
X-RAY DIFFRACTIONr_lrange_other5.77327.04373280
X-RAY DIFFRACTIONr_ncsr_local_group_10.0430.058552
X-RAY DIFFRACTIONr_ncsr_local_group_20.0450.058552
X-RAY DIFFRACTIONr_ncsr_local_group_30.0580.058487
X-RAY DIFFRACTIONr_ncsr_local_group_40.050.058551
X-RAY DIFFRACTIONr_ncsr_local_group_50.0490.058577
X-RAY DIFFRACTIONr_ncsr_local_group_60.0550.058556
X-RAY DIFFRACTIONr_ncsr_local_group_70.0640.058496
X-RAY DIFFRACTIONr_ncsr_local_group_80.050.058634
X-RAY DIFFRACTIONr_ncsr_local_group_90.0570.058568
X-RAY DIFFRACTIONr_ncsr_local_group_100.0510.058640
X-RAY DIFFRACTIONr_ncsr_local_group_110.0530.058639
X-RAY DIFFRACTIONr_ncsr_local_group_120.0580.058615
X-RAY DIFFRACTIONr_ncsr_local_group_130.0660.058531
X-RAY DIFFRACTIONr_ncsr_local_group_140.0470.058573
X-RAY DIFFRACTIONr_ncsr_local_group_150.0450.058627
X-RAY DIFFRACTIONr_ncsr_local_group_160.0470.058624
X-RAY DIFFRACTIONr_ncsr_local_group_170.0460.058635
X-RAY DIFFRACTIONr_ncsr_local_group_180.0660.058535
X-RAY DIFFRACTIONr_ncsr_local_group_190.0430.058611
X-RAY DIFFRACTIONr_ncsr_local_group_200.060.058568
X-RAY DIFFRACTIONr_ncsr_local_group_210.0570.058582
X-RAY DIFFRACTIONr_ncsr_local_group_220.0630.058560
X-RAY DIFFRACTIONr_ncsr_local_group_230.0510.058681
X-RAY DIFFRACTIONr_ncsr_local_group_240.0540.058684
X-RAY DIFFRACTIONr_ncsr_local_group_250.0630.058610
X-RAY DIFFRACTIONr_ncsr_local_group_260.0580.058488
X-RAY DIFFRACTIONr_ncsr_local_group_270.0720.058408
X-RAY DIFFRACTIONr_ncsr_local_group_280.0710.058503
Refine LS restraints NCS
Ens-IDDom-IDAuth asym-IDRefine-IDTypeRms dev position (Å)Weight position
11AX-RAY DIFFRACTIONLocal ncs0.043220.0501
12AX-RAY DIFFRACTIONLocal ncs0.043220.0501
23AX-RAY DIFFRACTIONLocal ncs0.044730.0501
24AX-RAY DIFFRACTIONLocal ncs0.044730.0501
35AX-RAY DIFFRACTIONLocal ncs0.058480.0501
36AX-RAY DIFFRACTIONLocal ncs0.058480.0501
47AX-RAY DIFFRACTIONLocal ncs0.049940.0501
48AX-RAY DIFFRACTIONLocal ncs0.049940.0501
59AX-RAY DIFFRACTIONLocal ncs0.048990.0501
510AX-RAY DIFFRACTIONLocal ncs0.048990.0501
611AX-RAY DIFFRACTIONLocal ncs0.055080.0501
612AX-RAY DIFFRACTIONLocal ncs0.055080.0501
713AX-RAY DIFFRACTIONLocal ncs0.064030.0501
714AX-RAY DIFFRACTIONLocal ncs0.064030.0501
815AX-RAY DIFFRACTIONLocal ncs0.050380.0501
816AX-RAY DIFFRACTIONLocal ncs0.050380.0501
917AX-RAY DIFFRACTIONLocal ncs0.056720.0501
918AX-RAY DIFFRACTIONLocal ncs0.056720.0501
1019AX-RAY DIFFRACTIONLocal ncs0.051370.0501
1020AX-RAY DIFFRACTIONLocal ncs0.051370.0501
1121AX-RAY DIFFRACTIONLocal ncs0.053270.0501
1122AX-RAY DIFFRACTIONLocal ncs0.053270.0501
1223AX-RAY DIFFRACTIONLocal ncs0.058190.0501
1224AX-RAY DIFFRACTIONLocal ncs0.058190.0501
1325AX-RAY DIFFRACTIONLocal ncs0.066050.0501
1326AX-RAY DIFFRACTIONLocal ncs0.066050.0501
1427AX-RAY DIFFRACTIONLocal ncs0.047470.0501
1428AX-RAY DIFFRACTIONLocal ncs0.047470.0501
1529AX-RAY DIFFRACTIONLocal ncs0.044630.0501
1530AX-RAY DIFFRACTIONLocal ncs0.044630.0501
1631AX-RAY DIFFRACTIONLocal ncs0.047220.0501
1632AX-RAY DIFFRACTIONLocal ncs0.047220.0501
1733AX-RAY DIFFRACTIONLocal ncs0.045760.0501
1734AX-RAY DIFFRACTIONLocal ncs0.045760.0501
1835AX-RAY DIFFRACTIONLocal ncs0.065640.0501
1836AX-RAY DIFFRACTIONLocal ncs0.065640.0501
1937AX-RAY DIFFRACTIONLocal ncs0.043380.0501
1938AX-RAY DIFFRACTIONLocal ncs0.043380.0501
2039AX-RAY DIFFRACTIONLocal ncs0.059680.0501
2040AX-RAY DIFFRACTIONLocal ncs0.059680.0501
2141AX-RAY DIFFRACTIONLocal ncs0.056580.0501
2142AX-RAY DIFFRACTIONLocal ncs0.056580.0501
2243AX-RAY DIFFRACTIONLocal ncs0.062640.0501
2244AX-RAY DIFFRACTIONLocal ncs0.062640.0501
2345AX-RAY DIFFRACTIONLocal ncs0.050750.0501
2346AX-RAY DIFFRACTIONLocal ncs0.050750.0501
2447AX-RAY DIFFRACTIONLocal ncs0.05440.0501
2448AX-RAY DIFFRACTIONLocal ncs0.05440.0501
2549AX-RAY DIFFRACTIONLocal ncs0.06260.0501
2550AX-RAY DIFFRACTIONLocal ncs0.06260.0501
2651AX-RAY DIFFRACTIONLocal ncs0.057610.0501
2652AX-RAY DIFFRACTIONLocal ncs0.057610.0501
2753AX-RAY DIFFRACTIONLocal ncs0.072010.0501
2754AX-RAY DIFFRACTIONLocal ncs0.072010.0501
2855AX-RAY DIFFRACTIONLocal ncs0.07130.0501
2856AX-RAY DIFFRACTIONLocal ncs0.07130.0501
LS refinement shell
Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkRefine-ID% reflection obs (%)
1.66-1.7030.2986990.27713910X-RAY DIFFRACTION91.4549
1.703-1.750.2837010.26314484X-RAY DIFFRACTION97.321
1.75-1.8010.2716980.24714066X-RAY DIFFRACTION97.4843
1.801-1.8560.2417120.22213688X-RAY DIFFRACTION97.6934
1.856-1.9170.2486870.21513303X-RAY DIFFRACTION97.798
1.917-1.9840.247230.19912770X-RAY DIFFRACTION97.8534
1.984-2.0590.2166680.19412406X-RAY DIFFRACTION98.0869
2.059-2.1430.2246240.18211962X-RAY DIFFRACTION98.1135
2.143-2.2380.1946120.17311525X-RAY DIFFRACTION98.5226
2.238-2.3470.1966240.16711030X-RAY DIFFRACTION98.5873
2.347-2.4730.1795810.1610447X-RAY DIFFRACTION98.8704
2.473-2.6230.195250.1629990X-RAY DIFFRACTION98.9275
2.623-2.8030.25070.1639369X-RAY DIFFRACTION99.1069
2.803-3.0270.1974970.1648714X-RAY DIFFRACTION99.31
3.027-3.3150.1864530.1718081X-RAY DIFFRACTION99.4175
3.315-3.7040.1823170.1587388X-RAY DIFFRACTION99.4322
3.704-4.2730.1543150.1416483X-RAY DIFFRACTION99.4878
4.273-5.2240.1452830.1255515X-RAY DIFFRACTION99.7076
5.224-7.3460.172480.1464262X-RAY DIFFRACTION99.7126
7.346-490.1411600.1482416X-RAY DIFFRACTION99.5748

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