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- PDB-2wlf: Crystallographic analysis of the polysialic acid O-acetyltransfer... -

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Basic information

Entry
Database: PDB / ID: 2wlf
TitleCrystallographic analysis of the polysialic acid O-acetyltransferase OatWY
ComponentsPOLYSIALIC ACID O-ACETYLTRANSFERASE
KeywordsTRANSFERASE / ENZYME / ACETYLTRANSFERASE / LEFT-HANDED BETA HELIX
Function / homology
Function and homology information


polysialic-acid O-acetyltransferase activity / protein homotrimerization / Transferases; Acyltransferases; Transferring groups other than aminoacyl groups / cytosol
Similarity search - Function
Hexapeptide repeat proteins / UDP N-Acetylglucosamine Acyltransferase; domain 1 / Hexapeptide repeat / Bacterial transferase hexapeptide (six repeats) / Trimeric LpxA-like superfamily / 3 Solenoid / Mainly Beta
Similarity search - Domain/homology
ACETYL COENZYME *A / ACETATE ION / Polysialic acid O-acetyltransferase
Similarity search - Component
Biological speciesNEISSERIA MENINGITIDIS SEROGROUP Y (bacteria)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 2.35 Å
AuthorsLee, H.J. / Rakic, B. / Gilbert, M. / Wakarchuk, W.W. / Withers, S.G. / Strynadka, N.C.J.
CitationJournal: J.Biol.Chem. / Year: 2009
Title: Structural and Kinetic Characterizations of the Polysialic Acid O-Acetyltransferase Oatwy from Neisseria Meningitidis.
Authors: Lee, H.J. / Rakic, B. / Gilbert, M. / Wakarchuk, W.W. / Withers, S.G. / Strynadka, N.C.J.
History
DepositionJun 23, 2009Deposition site: PDBE / Processing site: PDBE
Revision 1.0Jun 30, 2009Provider: repository / Type: Initial release
Revision 1.1Jul 13, 2011Group: Advisory / Version format compliance
Revision 1.2May 1, 2024Group: 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 / struct_ncs_dom_lim / struct_site
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 / _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 / _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

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Assembly

Deposited unit
A: POLYSIALIC ACID O-ACETYLTRANSFERASE
B: POLYSIALIC ACID O-ACETYLTRANSFERASE
C: POLYSIALIC ACID O-ACETYLTRANSFERASE
hetero molecules


Theoretical massNumber of molelcules
Total (without water)74,23416
Polymers71,1943
Non-polymers3,04013
Water4,540252
1


  • Idetical with deposited unit
  • defined by author&software
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area8820 Å2
ΔGint-44.6 kcal/mol
Surface area31160 Å2
MethodPQS
Unit cell
Length a, b, c (Å)78.855, 94.656, 100.829
Angle α, β, γ (deg.)90.00, 90.00, 90.00
Int Tables number16
Space group name H-MP222
Noncrystallographic symmetry (NCS)NCS domain:
IDEns-IDDetails
11A
21B
31C

NCS domain segments:

Component-ID: 1 / Ens-ID: 1 / Beg auth comp-ID: TYR / Beg label comp-ID: TYR / End auth comp-ID: TYR / End label comp-ID: TYR / Refine code: 6 / Auth seq-ID: 6 - 214 / Label seq-ID: 6 - 214

Dom-IDAuth asym-IDLabel asym-ID
1AA
2BB
3CC

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Components

#1: Protein POLYSIALIC ACID O-ACETYLTRANSFERASE


Mass: 23731.191 Da / Num. of mol.: 3 / Mutation: YES
Source method: isolated from a genetically manipulated source
Details: LEFT-HANDED BETA HELIX
Source: (gene. exp.) NEISSERIA MENINGITIDIS SEROGROUP Y (bacteria)
Strain: 406Y / Plasmid: PET-28 / Production host: ESCHERICHIA COLI (E. coli) / Strain (production host): DE3 / References: UniProt: Q93S40
#2: Chemical ChemComp-ACT / ACETATE ION


Mass: 59.044 Da / Num. of mol.: 3 / Source method: obtained synthetically / Formula: C2H3O2
#3: Chemical
ChemComp-EDO / 1,2-ETHANEDIOL / ETHYLENE GLYCOL


Mass: 62.068 Da / Num. of mol.: 7 / Source method: obtained synthetically / Formula: C2H6O2
#4: Chemical ChemComp-ACO / ACETYL COENZYME *A


Mass: 809.571 Da / Num. of mol.: 3 / Source method: obtained synthetically / Formula: C23H38N7O17P3S
#5: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 252 / Source method: isolated from a natural source / Formula: H2O
Compound detailsENGINEERED RESIDUE IN CHAIN A, ASN 67 TO ILE ENGINEERED RESIDUE IN CHAIN B, ASN 67 TO ILE ...ENGINEERED RESIDUE IN CHAIN A, ASN 67 TO ILE ENGINEERED RESIDUE IN CHAIN B, ASN 67 TO ILE ENGINEERED RESIDUE IN CHAIN C, ASN 67 TO ILE

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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: 2.65 Å3/Da / Density % sol: 54 %
Description: STARTING MODEL WAS MY NAI SOAKING SIRAS STRUCTURE SOLVED BY SHELX-CDE.
Crystal growpH: 4.6
Details: 100 MM SODIUM ACETATE, 2.25 M AMMONIUM ACETATE, pH 4.6

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: ALS / Beamline: 4.2.2 / Wavelength: 0.979
DetectorType: MBC NOIR-1 / Detector: CCD / Date: Jun 26, 2008 / Details: MIRRORS
RadiationMonochromator: DOUBLE CRYSTAL / Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.979 Å / Relative weight: 1
ReflectionResolution: 2.35→40 Å / Num. obs: 32150 / % possible obs: 100 % / Observed criterion σ(I): 3 / Redundancy: 4.8 % / Rmerge(I) obs: 0.1 / Net I/σ(I): 15
Reflection shellResolution: 2.35→2.48 Å / Redundancy: 4.9 % / Rmerge(I) obs: 0.44 / Mean I/σ(I) obs: 3.2 / % possible all: 100

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Processing

Software
NameVersionClassification
REFMAC5.5.0066refinement
MOSFLMdata reduction
SCALAdata scaling
PHASERphasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: IN-HOUSE STRUCTURE

Resolution: 2.35→40 Å / Cor.coef. Fo:Fc: 0.947 / Cor.coef. Fo:Fc free: 0.915 / SU B: 15.141 / SU ML: 0.168 / TLS residual ADP flag: LIKELY RESIDUAL / Cross valid method: THROUGHOUT / ESU R: 0.334 / ESU R Free: 0.231 / Stereochemistry target values: MAXIMUM LIKELIHOOD / Details: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS.
RfactorNum. reflection% reflectionSelection details
Rfree0.23094 1631 5.1 %RANDOM
Rwork0.18255 ---
obs0.18504 30518 99.96 %-
Solvent computationIon probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.4 Å / Solvent model: MASK
Displacement parametersBiso mean: 12.783 Å2
Baniso -1Baniso -2Baniso -3
1-2.37 Å20 Å20 Å2
2---1.02 Å20 Å2
3----1.35 Å2
Refinement stepCycle: LAST / Resolution: 2.35→40 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms4872 0 193 252 5317
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.0090.0225177
X-RAY DIFFRACTIONr_bond_other_d0.0010.024624
X-RAY DIFFRACTIONr_angle_refined_deg1.4681.9797032
X-RAY DIFFRACTIONr_angle_other_deg0.5423.00310713
X-RAY DIFFRACTIONr_dihedral_angle_1_deg5.85649
X-RAY DIFFRACTIONr_dihedral_angle_2_deg38.14725.735211
X-RAY DIFFRACTIONr_dihedral_angle_3_deg15.78515899
X-RAY DIFFRACTIONr_dihedral_angle_4_deg16.3931515
X-RAY DIFFRACTIONr_chiral_restr0.1570.2822
X-RAY DIFFRACTIONr_gen_planes_refined0.0030.025635
X-RAY DIFFRACTIONr_gen_planes_other00.02903
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_it0.3971.53166
X-RAY DIFFRACTIONr_mcbond_other0.0721.51319
X-RAY DIFFRACTIONr_mcangle_it0.76925152
X-RAY DIFFRACTIONr_mcangle_other
X-RAY DIFFRACTIONr_scbond_it1.19932011
X-RAY DIFFRACTIONr_scbond_other
X-RAY DIFFRACTIONr_scangle_it2.0584.51870
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

Ens-ID: 1 / Number: 3021 / Refine-ID: X-RAY DIFFRACTION

Dom-IDAuth asym-IDTypeRms dev position (Å)Weight position
1Aloose positional0.65
2Bloose positional0.565
3Cloose positional0.545
1Aloose thermal2.5210
2Bloose thermal2.410
3Cloose thermal2.110
LS refinement shellResolution: 2.35→2.411 Å / Total num. of bins used: 20
RfactorNum. reflection% reflection
Rfree0.304 125 -
Rwork0.226 2231 -
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)
17.01356.78173.615510.42356.6468.0701-0.15810.7150.4865-0.64370.37650.8743-0.3732-0.8682-0.21840.38530.1532-0.05920.4990.21750.337722.26446.3126-32.0933
20.4335-0.3048-0.48771.89480.61474.7651-0.00240.06710.0296-0.10260.0270.1577-0.065-0.3437-0.02460.13230.028-0.01750.16610.0360.143727.66660.681-36.0654
31.98721.34521.52774.4693-0.31172.3094-0.1037-0.00460.12-0.00940.02390.3080.2263-0.42530.07980.2568-0.1410.00610.3341-0.0290.220427.2858-6.6017-36.3922
40.8076-0.3887-0.58013.42581.10463.85370.00560.03370.0124-0.03090.0348-0.01220.06490.0664-0.04040.0735-0.0207-0.01830.160.00580.122332.3375-8.1372-31.7137
50.2764-0.3702-0.52751.23340.73982.2344-0.0083-0.028-0.037-0.0322-0.033-0.00860.23250.07380.04120.0839-0.0270.01980.13430.02170.104935.6379-16.0931-21.137
611.5502-2.2563-5.16565.29412.47669.2614-0.3041-0.6746-0.38420.19280.1868-0.43820.56830.7930.11740.20810.0461-0.00430.18410.040.171443.8812-22.8162-19.3336
74.9025-0.3087-1.60972.6551-0.18023.8823-0.0521-0.0985-0.27440.16020.0055-0.10560.47750.26630.04660.22080.03830.07770.08320.03030.175543.5196-29.289-30.4123
810.6408-4.1477-1.45914.7046-3.15416.0119-0.06651.13170.4374-0.1292-0.0497-0.31110.1114-0.0830.11620.2883-0.01280.16440.4940.11610.269649.5448-26.7343-44.0169
911.368-3.1168-2.012613.21066.937812.5738-0.38380.60340.10430.2480.30630.51221.0258-0.66740.07750.5313-0.1385-0.06470.50820.09870.34812.3561-30.5697-53.0532
105.95290.7511-1.8422.0837-1.13164.67190.08850.1557-0.1047-0.22840.00240.27640.7277-0.7612-0.09090.4075-0.2016-0.03970.2608-0.02060.164410.905-38.2369-47.9721
114.1866-0.1346-2.14620.4087-0.40174.19860.17980.0575-0.0244-0.3171-0.0274-0.00040.5347-0.3251-0.15250.402-0.16260.01890.2204-0.02110.177615.854-38.0693-43.9239
128.0652-0.0637-3.23145.29850.25448.08840.1290.2487-0.1765-0.2395-0.03240.04120.2683-0.3031-0.09660.3427-0.1583-0.01720.1521-0.02570.106319.3515-37.1529-40.9301
134.0344-0.1095-1.32261.1046-0.95213.9530.09480.0257-0.1725-0.1912-0.0202-0.08240.289-0.2342-0.07460.3732-0.16290.04070.1491-0.02880.16619.758-40.036-34.1775
143.07811.56-2.01961.0215-1.14732.4705-0.0682-0.0225-0.2433-0.17870.0116-0.18430.40610.05150.05660.3877-0.05530.0370.1173-0.0180.199832.6037-38.9668-33.154
152.11842.21-1.00623.6377-0.42692.3209-0.033-0.118-0.3956-0.1157-0.0308-0.35190.52780.19510.06390.4063-0.09630.01630.12020.0210.203325.1425-45.8346-20.0457
162.13022.7927-1.27453.855-1.49621.7666-0.36330.32820.4958-0.69760.47011.00970.307-0.6699-0.10680.6885-0.3198-0.25060.37160.1040.92918.4732-54.6081-18.5859
1721.51282.1522-12.250411.0369-15.808126.69290.20761.03650.39710.2158-1.0628-0.5711-0.3520.83640.85520.2468-0.2695-0.14880.7609-0.29631.2083-9.3803-23.657-15.3334
180.09630.64920.47514.99714.10393.68720.0628-0.02090.1920.2689-0.36280.8370.0115-0.430.30.252-0.04950.22720.43790.0650.7351-3.7483-15.9501-10.425
195.79434.42311.46549.5762.15223.40560.08330.01860.34120.4671-0.24440.45350.2362-0.31310.16110.1311-0.07040.08970.2741-0.0460.3278-1.4522-19.9551-9.8917
2012.48371.111-1.097211.3195-1.78317.9193-0.22960.4166-0.6064-0.76570.0838-0.01670.8514-0.27260.14580.1684-0.08090.02730.2960.02770.2084.029-20.9201-18.3526
212.56060.80010.29172.63210.18361.3919-0.0911-0.00360.06840.0860.06090.28580.3074-0.26880.03030.14-0.12050.05460.20720.02230.10478.6386-24.3334-9.3133
224.43221.9994-2.439611.62773.05924.7899-0.0555-0.1971-0.34330.42090.0672-0.13010.45660.0504-0.01170.3595-0.17350.05520.2701-0.01980.180213.6982-48.4771-14.608
231.11681.02110.37152.19861.17980.79020.0314-0.0183-0.09170.2115-0.0608-0.0040.2645-0.16420.02950.2547-0.10940.04790.19640.03340.128619.4734-30.2081-7.6737
241.68131.50680.34823.29370.58592.08560.0704-0.1385-0.09650.4396-0.0747-0.00940.2199-0.16370.00430.1241-0.02280.04290.16590.01680.079827.71-17.9696-1.8122
Refinement TLS group
IDRefine-IDRefine TLS-IDAuth asym-IDAuth seq-ID
1X-RAY DIFFRACTION1A6 - 11
2X-RAY DIFFRACTION2A12 - 38
3X-RAY DIFFRACTION3A39 - 53
4X-RAY DIFFRACTION4A54 - 93
5X-RAY DIFFRACTION5A94 - 169
6X-RAY DIFFRACTION6A170 - 176
7X-RAY DIFFRACTION7A177 - 200
8X-RAY DIFFRACTION8A201 - 214
9X-RAY DIFFRACTION9B5 - 9
10X-RAY DIFFRACTION10B10 - 44
11X-RAY DIFFRACTION11B45 - 65
12X-RAY DIFFRACTION12B66 - 81
13X-RAY DIFFRACTION13B82 - 116
14X-RAY DIFFRACTION14B117 - 159
15X-RAY DIFFRACTION15B160 - 197
16X-RAY DIFFRACTION16B198 - 214
17X-RAY DIFFRACTION17C6 - 10
18X-RAY DIFFRACTION18C11 - 26
19X-RAY DIFFRACTION19C27 - 45
20X-RAY DIFFRACTION20C46 - 51
21X-RAY DIFFRACTION21C52 - 118
22X-RAY DIFFRACTION22C119 - 127
23X-RAY DIFFRACTION23C128 - 168
24X-RAY DIFFRACTION24C169 - 214

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