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- PDB-3tzt: The structure of a protein in glycosyl transferase family 8 from ... -

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Basic information

Entry
Database: PDB / ID: 3tzt
TitleThe structure of a protein in glycosyl transferase family 8 from Anaerococcus prevotii.
ComponentsGlycosyl transferase family 8
KeywordsTRANSFERASE / Structural Genomics / PSI-Biology / Midwest Center for Structural Genomics / MCSG / putative glycosyl transferase / general stress protein
Function / homology
Function and homology information


glycosyltransferase activity
Similarity search - Function
: / Glycosyl transferase, family 8 / Glycosyl transferase family 8 / Spore Coat Polysaccharide Biosynthesis Protein SpsA; Chain A / Spore Coat Polysaccharide Biosynthesis Protein SpsA; Chain A / Nucleotide-diphospho-sugar transferases / Alpha-Beta Complex / Alpha Beta
Similarity search - Domain/homology
CITRIC ACID / Glycosyl transferase family 8
Similarity search - Component
Biological speciesAnaerococcus prevotii (bacteria)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MAD / Resolution: 2.1 Å
AuthorsCuff, M.E. / Tesar, C. / Bearden, J. / Joachimiak, A. / Midwest Center for Structural Genomics (MCSG)
CitationJournal: TO BE PUBLISHED
Title: The structure of a protein in glycosyl transferase family 8 from Anaerococcus prevotii.
Authors: Cuff, M.E. / Tesar, C. / Bearden, J. / Joachimiak, A.
History
DepositionSep 27, 2011Deposition site: RCSB / Processing site: RCSB
Revision 1.0Dec 7, 2011Provider: repository / Type: Initial release
Revision 1.1Oct 9, 2013Group: Structure summary
Revision 1.2Nov 8, 2017Group: Refinement description / Category: software
Revision 1.3Oct 16, 2024Group: Data collection / Database references ...Data collection / Database references / Derived calculations / Structure summary
Category: chem_comp_atom / chem_comp_bond ...chem_comp_atom / chem_comp_bond / database_2 / pdbx_entry_details / pdbx_modification_feature / 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 / _struct_conn.pdbx_dist_value / _struct_conn.pdbx_leaving_atom_flag / _struct_conn.pdbx_ptnr1_label_alt_id / _struct_conn.pdbx_ptnr2_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_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_comp_id / _struct_conn.ptnr2_label_seq_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: Glycosyl transferase family 8
B: Glycosyl transferase family 8
hetero molecules


Theoretical massNumber of molelcules
Total (without water)66,27410
Polymers65,6482
Non-polymers6278
Water4,738263
1


  • Idetical with deposited unit
  • defined by software
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area9350 Å2
ΔGint-35 kcal/mol
Surface area20880 Å2
MethodPISA
Unit cell
Length a, b, c (Å)42.498, 99.331, 122.794
Angle α, β, γ (deg.)90.00, 90.00, 90.00
Int Tables number19
Space group name H-MP212121
DetailsLikely the AB dimer in the AU.

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Components

#1: Protein Glycosyl transferase family 8


Mass: 32823.875 Da / Num. of mol.: 2
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Anaerococcus prevotii (bacteria) / Strain: DSM 20548 / Gene: Apre_0416 / Plasmid: pMCSG7 / Production host: Escherichia coli (E. coli) / Strain (production host): BL21 (DE3) magic / References: UniProt: C7RG54
#2: Chemical
ChemComp-EDO / 1,2-ETHANEDIOL / ETHYLENE GLYCOL


Mass: 62.068 Da / Num. of mol.: 7 / Source method: obtained synthetically / Formula: C2H6O2
#3: Chemical ChemComp-CIT / CITRIC ACID


Mass: 192.124 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: C6H8O7
#4: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 263 / Source method: isolated from a natural source / Formula: H2O
Has protein modificationY

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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: 1.97 Å3/Da / Density % sol: 37.69 %
Crystal growTemperature: 297 K / Method: vapor diffusion, sitting drop / pH: 5
Details: 0.2M Amonium citrate dibasic, 20% PEG 3350, VAPOR DIFFUSION, SITTING DROP, temperature 297K

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: APS / Beamline: 19-ID / Wavelength: 0.97918, 0.97929
DetectorType: ADSC QUANTUM 315r / Detector: CCD / Date: Dec 9, 2010
RadiationMonochromator: SAGITALLY FOCUSED Si(111) / Protocol: MAD / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelength
IDWavelength (Å)Relative weight
10.979181
20.979291
ReflectionRedundancy: 7.1 % / Av σ(I) over netI: 27.44 / Number: 224942 / Rmerge(I) obs: 0.062 / Χ2: 0.89 / D res high: 2.1 Å / D res low: 50 Å / Num. obs: 31606 / % possible obs: 99.8
Diffraction reflection shell
Highest resolution (Å)Lowest resolution (Å)% possible obs (%)IDRmerge(I) obsChi squaredRedundancy
5.75097.110.0370.8366.2
4.525.799.910.0380.7326.7
3.954.5210010.0450.9796.8
3.593.9599.910.0490.9977
3.333.5910010.0490.8437.1
3.143.3310010.0540.7847.2
2.983.1410010.0650.817.2
2.852.9810010.0730.8127.3
2.742.8510010.0950.8867.3
2.652.7410010.1140.9127.3
2.562.6510010.1430.8967.3
2.492.5610010.1720.9367.3
2.422.4910010.1920.9497.3
2.372.4210010.230.9417.3
2.312.3710010.2620.9277.2
2.262.3110010.3150.9397.3
2.222.2610010.360.9477.2
2.182.2210010.4270.8977.3
2.142.1899.910.510.9117.2
2.12.1499.910.5880.8977.1
ReflectionResolution: 2.1→50 Å / Num. all: 31606 / Num. obs: 31606 / % possible obs: 99.8 % / Observed criterion σ(I): -3 / Redundancy: 7.1 % / Biso Wilson estimate: 25.1 Å2 / Rmerge(I) obs: 0.062 / Χ2: 0.892 / Net I/σ(I): 10.4
Reflection shell
Resolution (Å)Redundancy (%)Rmerge(I) obsNum. unique allΧ2Diffraction-ID% possible all
2.1-2.147.10.58815890.897199.9
2.14-2.187.20.5115200.911199.9
2.18-2.227.30.42715520.8971100
2.22-2.267.20.3615430.9471100
2.26-2.317.30.31515540.9391100
2.31-2.377.20.26215840.9271100
2.37-2.427.30.2315390.9411100
2.42-2.497.30.19215570.9491100
2.49-2.567.30.17215740.9361100
2.56-2.657.30.14315670.8961100
2.65-2.747.30.11415580.9121100
2.74-2.857.30.09515750.8861100
2.85-2.987.30.07315670.8121100
2.98-3.147.20.06515770.811100
3.14-3.337.20.05415980.7841100
3.33-3.597.10.04915930.8431100
3.59-3.9570.04915980.997199.9
3.95-4.526.80.04516140.9791100
4.52-5.76.70.03816420.732199.9
5.7-506.20.03717050.836197.1

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Phasing

PhasingMethod: MAD
Phasing MADD res high: 2.1 Å / D res low: 50 Å / FOM : 0.347 / FOM acentric: 0.371 / FOM centric: 0.164 / Reflection: 30355 / Reflection acentric: 26744 / Reflection centric: 3611
Phasing MAD set

Highest resolution: 2.1 Å / Lowest resolution: 50 Å

IDR cullis acentricR cullis centricLoc acentricLoc centricPower acentricPower centricReflection acentricReflection centric
11.4910.30.300267443611
20.970.9437.753.20.380.31170642636
Phasing MAD set shell
IDResolution (Å)R cullis acentricR cullis centricLoc acentricLoc centricPower acentricPower centricReflection acentricReflection centric
112.98-501.0510.70.6007273
17.46-12.981.1410.90.700419182
15.23-7.461.5710.70.4001085304
14.03-5.231.0710.60.5002045419
13.28-4.031.210.40.3003289524
12.76-3.281.8910.40.2004866643
12.39-2.762.2510.20.1006726746
12.1-2.391.6210.10008242720
212.98-500.920.955.660.80.710.636868
27.46-12.980.910.8954.957.30.610.55417178
25.23-7.460.930.873847.60.740.551085304
24.03-5.230.960.9450.166.20.450.322045419
23.28-4.030.980.9544.662.50.360.243289524
22.76-3.280.980.9633.545.70.350.224866643
22.39-2.760.990.9830.842.90.260.155294500
22.1-2.3900000000
Phasing MAD set site

Atom type symbol: Se

IDB isoFract xFract yFract zOccupancyOccupancy iso
121.73850.2070.40.1427.3370
227.15980.0370.3690.1878.4170
319.1715-0.3220.4090.1726.1560
422.9331-0.1510.4470.1316.6790
529.8820.2570.40.0175.0670
621.0161-0.4380.2610.1834.7990
734.8677-0.3120.2470.1614.3820
823.8419-0.2140.2830.2424.4980
933.91470.0840.4920.0464.0930
1041.07330.1220.3840.0153.720
1139.1673-0.3670.1560.384.3860
1266.74560.2720.393-0.0952.0320
1315.74670.2060.3990.1414.358-0.192
1423.00230.0370.3680.1884.795-0.243
1516.7049-0.3240.4070.1723.432-0.17
1623.6521-0.1520.4470.1324.206-0.226
1741.57240.2550.3980.0172.284-0.276
1822.3385-0.4370.2610.182.438-0.243
1931.451-0.3130.2470.1612.52-0.168
2015.7014-0.2120.2820.2412.916-0.114
2143.62280.0810.4920.0463.041-0.231
22101.48860.1240.3840.0163.095-0.206
2339.662-0.3620.1530.3822.698-0.193
2451.3640.2660.392-0.0961.045-0.116
Phasing MAD shell
Resolution (Å)FOM FOM acentricFOM centricReflectionReflection acentricReflection centric
12.98-500.4430.5740.3141457273
7.46-12.980.5390.6420.303601419182
5.23-7.460.6230.6960.36413891085304
4.03-5.230.5480.6090.24924642045419
3.28-4.030.5350.5850.22138133289524
2.76-3.280.4840.5230.19355094866643
2.39-2.760.3050.330.0874726726746
2.1-2.390.1050.114089628242720
Phasing dmMethod: Solvent flattening and Histogram matching / Reflection: 30355
Phasing dm shell
Resolution (Å)Delta phi finalFOM Reflection
8.43-10064.60.52508
6.74-8.4351.80.866506
5.8-6.7447.70.893566
5.17-5.843.80.916639
4.71-5.1749.40.917694
4.35-4.7148.60.936766
4.06-4.35550.922816
3.83-4.0652.90.923848
3.63-3.8350.70.919914
3.46-3.63520.908964
3.31-3.4657.10.9995
3.18-3.3154.20.8991031
3.06-3.1856.50.8841077
2.95-3.0653.90.8851109
2.86-2.9556.40.8891135
2.77-2.8659.30.8671178
2.69-2.77610.8541230
2.62-2.6962.50.8431240
2.55-2.6264.90.8081279
2.49-2.5564.60.8231300
2.44-2.4970.10.831378
2.38-2.4472.60.8321351
2.33-2.3875.10.8111387
2.28-2.33810.8181428
2.24-2.2878.50.8131375
2.2-2.2482.40.8131414
2.16-2.2830.7741324
2.1-2.16860.7391903

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Processing

Software
NameVersionClassificationNB
DENZOdata reduction
SCALEPACKdata scaling
MLPHAREphasing
DM6.1phasing
REFMACrefinement
PDB_EXTRACT3.1data extraction
SBC-Collectdata collection
HKL-3000data reduction
HKL-3000data scaling
HKL-3000phasing
SHELXDphasing
SHELXEmodel building
SOLVEphasing
RESOLVEphasing
ARP/wARPmodel building
CCP4phasing
Omodel building
COOphasing
RefinementMethod to determine structure: MAD / Resolution: 2.1→37.84 Å / Cor.coef. Fo:Fc: 0.958 / Cor.coef. Fo:Fc free: 0.935 / WRfactor Rfree: 0.2122 / WRfactor Rwork: 0.1592 / Occupancy max: 1 / Occupancy min: 0.4 / FOM work R set: 0.9134 / SU B: 7.167 / SU ML: 0.096 / SU R Cruickshank DPI: 0.2045 / SU Rfree: 0.17 / Cross valid method: THROUGHOUT / σ(F): 0 / ESU R Free: 0.17
Stereochemistry target values: MAXIMUM LIKELIHOOD WITH PHASES
Details: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS U VALUES : WITH TLS ADDED
RfactorNum. reflection% reflectionSelection details
Rfree0.2059 1518 5 %RANDOM
Rwork0.1594 ---
all0.1617 30241 --
obs0.1617 30241 96.88 %-
Solvent computationIon probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.2 Å / Solvent model: MASK
Displacement parametersBiso max: 113.89 Å2 / Biso mean: 35.7841 Å2 / Biso min: 11.28 Å2
Baniso -1Baniso -2Baniso -3
1-0.22 Å20 Å20 Å2
2---0.09 Å20 Å2
3----0.13 Å2
Refinement stepCycle: LAST / Resolution: 2.1→37.84 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms3839 0 41 263 4143
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.0150.024028
X-RAY DIFFRACTIONr_bond_other_d0.0010.022775
X-RAY DIFFRACTIONr_angle_refined_deg1.591.995451
X-RAY DIFFRACTIONr_angle_other_deg0.96336736
X-RAY DIFFRACTIONr_dihedral_angle_1_deg6.2585475
X-RAY DIFFRACTIONr_dihedral_angle_2_deg37.74823.568199
X-RAY DIFFRACTIONr_dihedral_angle_3_deg16.20715712
X-RAY DIFFRACTIONr_dihedral_angle_4_deg20.6381529
X-RAY DIFFRACTIONr_chiral_restr0.0980.2595
X-RAY DIFFRACTIONr_gen_planes_refined0.0070.0214390
X-RAY DIFFRACTIONr_gen_planes_other0.0010.02843
LS refinement shellResolution: 2.1→2.154 Å / Total num. of bins used: 20
RfactorNum. reflection% reflection
Rfree0.234 82 -
Rwork0.19 1598 -
all-1680 -
obs--78.14 %
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)
10.56410.5366-0.29451.4463-0.77391.64310.0567-0.05-0.07460.0345-0.0842-0.00320.024-0.00410.02750.03180.0356-0.0020.0851-0.00780.0965-2.969829.085423.2813
20.75580.5379-0.3311.5332-0.56751.11960.0270.09890.1187-0.22490.03030.1255-0.1056-0.1088-0.05730.12110.0802-0.00140.10130.00310.0935-3.686743.39298.5227
Refinement TLS group
IDRefine-IDRefine TLS-IDAuth asym-IDAuth seq-ID
1X-RAY DIFFRACTION1A-10 - 9999
2X-RAY DIFFRACTION2B-10 - 9999

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