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- PDB-4do7: Crystal structure of an amidohydrolase (cog3618) from burkholderi... -

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Basic information

Entry
Database: PDB / ID: 4do7
TitleCrystal structure of an amidohydrolase (cog3618) from burkholderia multivorans (target efi-500235) with bound zn, space group c2
ComponentsAmidohydrolase 2
KeywordsHYDROLASE / Amidohydrolase / enzyme function initiative / EFI / Structural Genomics / TIM-barrel fold / putative lactonase
Function / homology
Function and homology information


L-fucono-1,5-lactonase / fucose catabolic process / hydrolase activity / metal ion binding
Similarity search - Function
Amidohydrolase / Amidohydrolase-related / Metal-dependent hydrolases / Metal-dependent hydrolase / TIM Barrel / Alpha-Beta Barrel / Alpha Beta
Similarity search - Domain/homology
L-fucono-1,5-lactonase / Putative amidohydrolase
Similarity search - Component
Biological speciesBurkholderia multivorans (bacteria)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 1.7 Å
AuthorsVetting, M.W. / Toro, R. / Bhosle, R. / Wasserman, S.R. / Morisco, L.L. / Sojitra, S. / Seidel, R.D. / Hillerich, B. / Washington, E. / Scott Glenn, A. ...Vetting, M.W. / Toro, R. / Bhosle, R. / Wasserman, S.R. / Morisco, L.L. / Sojitra, S. / Seidel, R.D. / Hillerich, B. / Washington, E. / Scott Glenn, A. / Chowdhury, S. / Evans, B. / Hammonds, J. / Al Obaidi, N.F. / Zencheck, W.D. / Imker, H.J. / Gerlt, J.A. / Raushel, F.M. / Almo, S.C. / Enzyme Function Initiative (EFI)
CitationJournal: to be published
Title: Crystal structure of an amidohydrolase (cog3618) from burkholderia multivorans (target efi-500235) with bound zn, space group c2
Authors: Vetting, M.W. / Toro, R. / Bhosle, R. / Wasserman, S.R. / Morisco, L.L. / Sojitra, S. / Seidel, R.D. / Hillerich, B. / Washington, E. / Scott Glenn, A. / Chowdhury, S. / Evans, B. / ...Authors: Vetting, M.W. / Toro, R. / Bhosle, R. / Wasserman, S.R. / Morisco, L.L. / Sojitra, S. / Seidel, R.D. / Hillerich, B. / Washington, E. / Scott Glenn, A. / Chowdhury, S. / Evans, B. / Hammonds, J. / Al Obaidi, N.F. / Zencheck, W.D. / Imker, H.J. / Gerlt, J.A. / Raushel, F.M. / Almo, S.C. / Enzyme Function Initiative (EFI)
History
DepositionFeb 9, 2012Deposition site: RCSB / Processing site: RCSB
Revision 1.0Feb 29, 2012Provider: repository / Type: Initial release
Revision 1.1Jan 31, 2018Group: Database references / Category: citation_author / Item: _citation_author.name
Revision 1.2Sep 13, 2023Group: 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 / pdbx_struct_conn_angle / struct_conn / struct_ref_seq_dif / struct_site
Item: _database_2.pdbx_DOI / _database_2.pdbx_database_accession ..._database_2.pdbx_DOI / _database_2.pdbx_database_accession / _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_seq_id / _pdbx_struct_conn_angle.ptnr2_label_asym_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 / _struct_site.pdbx_auth_asym_id / _struct_site.pdbx_auth_comp_id / _struct_site.pdbx_auth_seq_id

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: Amidohydrolase 2
B: Amidohydrolase 2
hetero molecules


Theoretical massNumber of molelcules
Total (without water)68,1485
Polymers67,9212
Non-polymers2273
Water9,152508
1
A: Amidohydrolase 2
hetero molecules


Theoretical massNumber of molelcules
Total (without water)34,0262
Polymers33,9601
Non-polymers651
Water181
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
2
B: Amidohydrolase 2
hetero molecules


Theoretical massNumber of molelcules
Total (without water)34,1223
Polymers33,9601
Non-polymers1612
Water181
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Unit cell
Length a, b, c (Å)103.332, 76.728, 82.862
Angle α, β, γ (deg.)90.000, 117.670, 90.000
Int Tables number5
Space group name H-MC121
Components on special symmetry positions
IDModelComponents
11B-771-

HOH

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Components

#1: Protein Amidohydrolase 2 / Putative amidohydrolase


Mass: 33960.367 Da / Num. of mol.: 2
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Burkholderia multivorans (bacteria) / Strain: ATCC 17616 / Gene: Bmul_3602, BMULJ_04915 / Plasmid: pET / Production host: Escherichia coli (E. coli) / Strain (production host): BL21(DE3) / References: UniProt: A9ANE4, UniProt: A0A0H3KNC4*PLUS
#2: Chemical ChemComp-ZN / ZINC ION


Mass: 65.409 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: Zn
#3: Chemical ChemComp-SO4 / SULFATE ION


Mass: 96.063 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: SO4
#4: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 508 / 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: 2.14 Å3/Da / Density % sol: 42.57 %
Crystal growTemperature: 298 K / Method: sitting drop vapor diffuction / pH: 7.5
Details: Protein (10 mM Hepes, pH 7.8, 150 mM NaCl, 10% glycerol, 0.5 mM ZnCl, 0.5 mM Fuconate 1,4-lactone; Reservoir (1.26 M AmmSO4, 100 mM HEPES pH 7.5); Cryoprotection (Reservoir, + 20% glycerol), ...Details: Protein (10 mM Hepes, pH 7.8, 150 mM NaCl, 10% glycerol, 0.5 mM ZnCl, 0.5 mM Fuconate 1,4-lactone; Reservoir (1.26 M AmmSO4, 100 mM HEPES pH 7.5); Cryoprotection (Reservoir, + 20% glycerol), sitting drop vapor diffuction, temperature 298K

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: APS / Beamline: 31-ID / Wavelength: 0.9793 Å
DetectorType: RAYONIX MX225HE / Detector: CCD / Date: Nov 15, 2011 / Details: MIRRORS
RadiationMonochromator: GRAPHITE / Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.9793 Å / Relative weight: 1
ReflectionResolution: 1.7→73.388 Å / Num. all: 62061 / Num. obs: 62061 / % possible obs: 98.5 % / Observed criterion σ(F): 0 / Observed criterion σ(I): 0 / Redundancy: 3.5 % / Rsym value: 0.062 / Net I/σ(I): 10.7
Reflection shell

Diffraction-ID: 1

Resolution (Å)Redundancy (%)Rmerge(I) obsMean I/σ(I) obsNum. measured allNum. unique allRsym value% possible all
1.7-1.793.30.39922948090520.39998.9
1.79-1.93.40.2662.92897385910.26699.4
1.9-2.033.50.1764.32823081000.17699.5
2.03-2.193.60.1166.32720475900.11699.6
2.19-2.43.70.0887.92548769690.08899.3
2.4-2.693.70.0792310462630.0798.7
2.69-3.13.70.05610.32011854530.05697.4
3.1-3.83.70.04611.41687145750.04696.2
3.8-5.383.70.03813.81300135290.03895.8
5.38-40.3833.60.03615.3698819390.03694.3

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Processing

Software
NameVersionClassificationNB
SCALA3.3.16data scaling
PHENIX1.7.3_928refinement
PDB_EXTRACT3.1data extraction
MOSFLMdata reduction
PHENIXphasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: PDB ENTRY 4DNM
Resolution: 1.7→30.626 Å / Occupancy max: 1 / Occupancy min: 0.38 / FOM work R set: 0.8429 / SU ML: 0.15 / σ(F): 1.34 / Phase error: 22.54 / Stereochemistry target values: ML
RfactorNum. reflection% reflectionSelection details
Rfree0.2091 3147 5.07 %RANDOM
Rwork0.176 ---
obs0.1777 62011 98.41 %-
all-62011 --
Solvent computationShrinkage radii: 0.98 Å / VDW probe radii: 1.2 Å / Solvent model: FLAT BULK SOLVENT MODEL / Bsol: 59.767 Å2 / ksol: 0.4 e/Å3
Displacement parametersBiso max: 74.25 Å2 / Biso mean: 25.8051 Å2 / Biso min: 9.23 Å2
Baniso -1Baniso -2Baniso -3
1-0.5635 Å20 Å22.095 Å2
2---1.1918 Å20 Å2
3---0.6283 Å2
Refinement stepCycle: LAST / Resolution: 1.7→30.626 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms4482 0 7 508 4997
Refine LS restraints
Refine-IDTypeDev idealNumber
X-RAY DIFFRACTIONf_bond_d0.0074677
X-RAY DIFFRACTIONf_angle_d1.0216372
X-RAY DIFFRACTIONf_chiral_restr0.072674
X-RAY DIFFRACTIONf_plane_restr0.005841
X-RAY DIFFRACTIONf_dihedral_angle_d12.6611650
LS refinement shell

Refine-ID: X-RAY DIFFRACTION / Total num. of bins used: 22

Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkNum. reflection all% reflection obs (%)
1.7-1.72660.28231480.23542649279798
1.7266-1.75490.28021470.22772652279999
1.7549-1.78510.2721380.21292711284999
1.7851-1.81760.2481440.21062643278799
1.8176-1.85250.2591420.20012715285799
1.8525-1.89030.2231310.1912679281099
1.8903-1.93140.23341390.19342686282599
1.9314-1.97640.20411680.17532684285299
1.9764-2.02580.20691290.17727192848100
2.0258-2.08050.22551640.179927032867100
2.0805-2.14170.20851470.17932688283599
2.1417-2.21090.20621610.17112691285299
2.2109-2.28990.22111350.1732681281699
2.2899-2.38150.21391600.17112692285299
2.3815-2.48980.19381240.17752716284099
2.4898-2.6210.23431230.1752680280399
2.621-2.78520.2081310.1842681281298
2.7852-30.20991430.18712644278797
3-3.30160.22581380.17042641277997
3.3016-3.77860.1571430.15752645278896
3.7786-4.75780.17381490.1432595274495
4.7578-30.63080.22561430.19032669281295
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)
12.09120.6982-0.16342.5439-0.49432.5255-0.0495-0.0029-0.15750.0886-0.0148-0.14920.03430.01320.07990.06770.0059-0.00430.08470.00340.086817.2091-7.177429.5314
22.0537-0.0401-0.07851.7620.0463.3749-0.0442-0.5296-0.0070.5530.0677-0.25540.06460.3271-0.04460.23240.0358-0.0730.21320.00030.146221.5655-1.485743.7413
32.44170.2938-0.99613.0922-0.00523.1442-0.019-0.5485-0.19230.7014-0.12260.03220.2560.06550.1880.28350.02790.02490.1646-0.00360.106210.3195-2.190645.7427
41.7412-0.38160.10772.29740.38662.26170.0248-0.4477-0.30870.66620.01790.4940.1995-0.42780.06440.16770.02930.10320.137-0.06420.11113.0351.9842.6912
52.58051.62770.0821.82490.5441.66840.01520.05780.04340.2717-0.23730.703-0.0138-0.53110.21470.15250.05010.03560.2143-0.07370.3342-4.32425.632635.6453
61.4184-0.3103-0.11931.2947-0.19551.49070.0590.171-0.092-0.3935-0.19140.5253-0.1005-0.4320.05350.13540.0429-0.07840.2007-0.05670.2081.4022-1.765725.1368
71.4229-1.0862-0.93293.0512-0.42311.37420.1290.22990.2019-0.5714-0.09820.1215-0.4587-0.19540.00780.21360.0465-0.01090.12680.0190.14778.21998.43923.037
81.6143-0.3952-0.44254.93220.86622.48-0.03490.099-0.2155-0.0357-0.14210.25710.2251-0.07280.15830.08460.01120.00590.1041-0.01440.110817.2314-29.05794.7451
91.73710.2632-0.34861.73080.41461.9432-0.09740.2885-0.0849-0.1790.03020.1540.0936-0.25670.03230.0743-0.0182-0.03020.1272-0.03340.0911.9851-23.28230.5217
103.3304-0.9198-1.14653.45130.41433.0965-0.05590.3647-0.1754-0.4690.1001-0.09110.04150.0344-0.02540.1916-0.0414-0.00310.1439-0.0460.095922.2002-22.4082-9.1149
111.67940.4459-0.72531.85830.15532.3327-0.05760.3611-0.205-0.33630.188-0.2808-0.09480.1943-0.06460.1341-0.02710.04910.1334-0.04740.1129.143-18.166-6.4166
124.7349-4.59052.90467.4624-2.33226.0791-0.066-0.22720.0566-0.072-0.0199-1.1196-0.24830.94240.20760.0959-0.02380.02570.3057-0.02480.335241.6969-17.30941.5563
131.32870.6009-0.75912.4217-0.04261.6923-0.0185-0.2199-0.08880.1860.0321-0.42480.0430.40690.02970.09250.0259-0.04020.1718-0.02580.154931.1863-20.62859.7713
141.10711.0648-0.98213.79430.40652.14090.1694-0.17630.09290.3070.0138-0.098-0.05760.285-0.12680.0807-0.00980.0130.1089-0.04270.128124.8875-11.068913.0553
Refinement TLS group
IDRefine-IDRefine TLS-IDSelection detailsAuth asym-IDAuth seq-ID
1X-RAY DIFFRACTION1chain 'A' and (resseq 3:59)A3 - 59
2X-RAY DIFFRACTION2chain 'A' and (resseq 60:96)A60 - 96
3X-RAY DIFFRACTION3chain 'A' and (resseq 97:121)A97 - 121
4X-RAY DIFFRACTION4chain 'A' and (resseq 122:180)A122 - 180
5X-RAY DIFFRACTION5chain 'A' and (resseq 181:202)A181 - 202
6X-RAY DIFFRACTION6chain 'A' and (resseq 203:253)A203 - 253
7X-RAY DIFFRACTION7chain 'A' and (resseq 254:289)A254 - 289
8X-RAY DIFFRACTION8chain 'B' and (resseq 3:27)B3 - 27
9X-RAY DIFFRACTION9chain 'B' and (resseq 28:96)B28 - 96
10X-RAY DIFFRACTION10chain 'B' and (resseq 97:121)B97 - 121
11X-RAY DIFFRACTION11chain 'B' and (resseq 122:176)B122 - 176
12X-RAY DIFFRACTION12chain 'B' and (resseq 180:193)B180 - 193
13X-RAY DIFFRACTION13chain 'B' and (resseq 194:253)B194 - 253
14X-RAY DIFFRACTION14chain 'B' and (resseq 254:289)B254 - 289

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