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- PDB-3d0q: Crystal structure of calG3 from Micromonospora echinospora determ... -

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Basic information

Entry
Database: PDB / ID: 3d0q
TitleCrystal structure of calG3 from Micromonospora echinospora determined in space group I222
ComponentsProtein CalG3
KeywordsTRANSFERASE / Calicheamicin synthesis / glycosyltransferase / enediyne antibiotic
Function / homology
Function and homology information


UDP-glycosyltransferase activity / glucosyltransferase activity / antibiotic biosynthetic process
Similarity search - Function
: / Erythromycin biosynthesis protein CIII-like, N-terminal domain / Erythromycin biosynthesis protein CIII-like, central / Erythromycin biosynthesis protein CIII-like, C-terminal domain / UDP-glucuronosyl/UDP-glucosyltransferase / Glycogen Phosphorylase B; / Rossmann fold / 3-Layer(aba) Sandwich / Alpha Beta
Similarity search - Domain/homology
Biological speciesMicromonospora echinospora (bacteria)
MethodX-RAY DIFFRACTION / SYNCHROTRON / SAD / Resolution: 2.79 Å
AuthorsBitto, E. / Singh, S. / Bingman, C.A. / Wesenberg, G.E. / Phillips Jr., G.N.
CitationJournal: Chem.Biol. / Year: 2008
Title: Biochemical and structural insights of the early glycosylation steps in calicheamicin biosynthesis.
Authors: Zhang, C. / Bitto, E. / Goff, R.D. / Singh, S. / Bingman, C.A. / Griffith, B.R. / Albermann, C. / Phillips, G.N. / Thorson, J.S.
History
DepositionMay 2, 2008Deposition site: RCSB / Processing site: RCSB
Revision 1.0Jun 24, 2008Provider: repository / Type: Initial release
Revision 1.1Jul 13, 2011Group: Advisory / Version format compliance
Revision 1.2Oct 25, 2017Group: Refinement description / Category: software

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: Protein CalG3
B: Protein CalG3
hetero molecules


Theoretical massNumber of molelcules
Total (without water)87,1944
Polymers86,7762
Non-polymers4192
Water81145
1


  • Idetical with deposited unit
  • defined by author&software
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area3310 Å2
ΔGint-18.5 kcal/mol
Surface area29870 Å2
MethodPISA
Unit cell
Length a, b, c (Å)106.654, 119.267, 155.942
Angle α, β, γ (deg.)90.000, 90.000, 90.000
Int Tables number23
Space group name H-MI222
Noncrystallographic symmetry (NCS)NCS domain:
IDEns-IDDetails
11A
21B
12A
22B
13A
23B

NCS domain segments:

Component-ID: 1 / Refine code: 2

Dom-IDEns-IDBeg label comp-IDEnd label comp-IDAuth asym-IDLabel asym-IDAuth seq-IDLabel seq-ID
11HISSERAA-2 - 5520 - 77
21HISSERBB-2 - 5520 - 77
12THRARGAA76 - 21098 - 232
22THRARGBB76 - 21098 - 232
13LEUILEAA259 - 374281 - 396
23LEUILEBB259 - 374281 - 396

NCS ensembles :
ID
1
2
3

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Components

#1: Protein Protein CalG3


Mass: 43387.977 Da / Num. of mol.: 2
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Micromonospora echinospora (bacteria) / Strain: LL6600 / Gene: calG3 / Plasmid: pET16b / Production host: Escherichia coli (E. coli) / Strain (production host): BL21(DE3) / References: UniProt: Q8KND7
#2: Chemical ChemComp-MPO / 3[N-MORPHOLINO]PROPANE SULFONIC ACID / MOPS


Mass: 209.263 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: C7H15NO4S / Comment: pH buffer*YM
#3: Water ChemComp-HOH / water / Water


Mass: 18.015 Da / Num. of mol.: 45 / 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: 2.86 Å3/Da / Density % sol: 56.95 %
Crystal growTemperature: 277 K / Method: vapor diffusion, hanging drop
Details: Protein solution (10 mg/mL Se-Met protein, 0.050 M Sodium chloride, 0.010 M Tris-HCl pH 7.5) mixed in a 1:1 ratio with the Well solution (16% PEG 4000, 0.20 M Tri-ammonium citrate, 0.10 M ...Details: Protein solution (10 mg/mL Se-Met protein, 0.050 M Sodium chloride, 0.010 M Tris-HCl pH 7.5) mixed in a 1:1 ratio with the Well solution (16% PEG 4000, 0.20 M Tri-ammonium citrate, 0.10 M MOPS pH 7.0). Cryoprotected in well solution containing up to 20% (v/v) Ethylene glycol, VAPOR DIFFUSION, HANGING DROP, temperature 277K

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: APS / Beamline: 23-ID-D / Wavelength: 0.97918 Å
DetectorType: MARMOSAIC 300 mm CCD / Detector: CCD / Date: Mar 26, 2006 / Details: Adjustable focusing mirrors in K-B geometry
RadiationMonochromator: Si(111) Double crystal / Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.97918 Å / Relative weight: 1
ReflectionResolution: 2.79→27 Å / Num. obs: 24883 / % possible obs: 99.2 % / Redundancy: 7.3 % / Rmerge(I) obs: 0.121 / Χ2: 1.125 / Net I/σ(I): 9.819
Reflection shell

Diffraction-ID: 1

Resolution (Å)Redundancy (%)Rmerge(I) obsMean I/σ(I) obsNum. unique allΧ2% possible all
2.79-2.96.30.4264.04623420.95294.9
2.9-3.0270.35424230.92598.2
3.02-3.157.40.27424491.00199.2
3.15-3.327.50.21524930.98699.9
3.32-3.537.50.17324751.10199.9
3.53-3.87.50.12724881.167100
3.8-4.187.50.09625071.08100
4.18-4.787.40.08125161.282100
4.78-6.017.40.07725411.277100
6.01-277.10.05926491.414100

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Phasing

PhasingMethod: SAD
Phasing MAD set

R cullis centric: 0 / Highest resolution: 3.2 Å / Lowest resolution: 26.66 Å / Power centric: 0

IDR cullis acentricPower acentricReflection acentricReflection centric
ISO_100149221808
ANO_10.7891.165149210
Phasing MAD set shell

R cullis centric: 0 / Power centric: 0

IDResolution (Å)R cullis acentricPower acentricReflection acentricReflection centric
ISO_112.68-26.660018394
ISO_19.52-12.680030691
ISO_17.94-9.520038388
ISO_16.96-7.940045794
ISO_16.27-6.960052987
ISO_15.75-6.270059190
ISO_15.34-5.750062089
ISO_15.01-5.340069690
ISO_14.73-5.010072490
ISO_14.49-4.730076092
ISO_14.29-4.490080387
ISO_14.11-4.290085493
ISO_13.95-4.110089293
ISO_13.81-3.950091987
ISO_13.69-3.810095997
ISO_13.57-3.690099290
ISO_13.47-3.5700101893
ISO_13.37-3.4700103985
ISO_13.28-3.3700108990
ISO_13.2-3.2800110888
ANO_112.68-26.660.4782.7531830
ANO_19.52-12.680.6332.13060
ANO_17.94-9.520.5432.3483830
ANO_16.96-7.940.5262.4934570
ANO_16.27-6.960.5222.3285290
ANO_15.75-6.270.5362.275910
ANO_15.34-5.750.5842.0916200
ANO_15.01-5.340.6541.7826960
ANO_14.73-5.010.6941.5877240
ANO_14.49-4.730.7171.4777600
ANO_14.29-4.490.7061.3718030
ANO_14.11-4.290.7891.1598540
ANO_13.95-4.110.8351.0278920
ANO_13.81-3.950.8720.8549180
ANO_13.69-3.810.8750.8179590
ANO_13.57-3.690.8830.7399920
ANO_13.47-3.570.9240.67210180
ANO_13.37-3.470.9240.58310390
ANO_13.28-3.370.9380.52810890
ANO_13.2-3.280.9480.47411080
Phasing MAD set site
IDCartn x (Å)Cartn y (Å)Cartn z (Å)Atom type symbolB isoOccupancy
1-83.921-4.388-9.593SE57.281.86
2-75.384-16.946-19.986SE62.282.36
3-83.251-23.945-47.163SE45.242.05
4-88.811-34.604-61.42SE36.711.66
5-85.026-9.618-58.701SE66.622.1
6-70.754-10.6143.104SE92.922.17
7-68.768-24.159-4.106SE38.551.6
8-82.293-41.125-53.613SE60.551.76
9-95.763-22.412-67.923SE78.581.8
10-66.548-33.661-7.477SE102.581.8
Phasing dmMethod: Solvent flattening and Histogram matching / Reflection: 16729
Phasing dm shell
Resolution (Å)Delta phi finalFOM Reflection
10.43-10075.70.631508
8.31-10.4364.80.838501
7.24-8.3166.10.847501
6.56-7.24660.834519
6.08-6.5666.20.814511
5.71-6.08670.842515
5.41-5.7166.10.868512
5.17-5.4165.50.862510
4.95-5.1759.10.887553
4.76-4.9560.20.88567
4.59-4.7661.30.887571
4.44-4.5961.30.892604
4.3-4.4461.80.906631
4.17-4.362.70.88651
4.06-4.1762.60.895663
3.95-4.0669.40.889669
3.85-3.9567.70.872683
3.76-3.8570.10.86728
3.68-3.76700.864734
3.6-3.6872.30.843731
3.52-3.672.50.856766
3.45-3.5276.80.836770
3.38-3.4577.80.823776
3.32-3.3877.60.755824
3.26-3.3276.60.746796
3.2-3.2680.70.692935

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Processing

Software
NameVersionClassificationNB
DENZOdata reduction
SCALEPACKdata scaling
SHARPphasing
DM5phasing
REFMAC5.2.0019refinement
PDB_EXTRACT3data extraction
MAR345CCDdata collection
RefinementMethod to determine structure: SAD / Resolution: 2.79→26.75 Å / Cor.coef. Fo:Fc: 0.939 / Cor.coef. Fo:Fc free: 0.893 / WRfactor Rfree: 0.239 / WRfactor Rwork: 0.184 / SU B: 28.63 / SU ML: 0.259 / TLS residual ADP flag: LIKELY RESIDUAL / Cross valid method: THROUGHOUT / σ(F): 0 / ESU R: 1.881 / ESU R Free: 0.342 / Stereochemistry target values: MAXIMUM LIKELIHOOD / Details: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS
RfactorNum. reflection% reflectionSelection details
Rfree0.243 1270 5.104 %RANDOM
Rwork0.187 ---
obs0.19 24883 98.883 %-
Solvent computationIon probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.4 Å / Solvent model: MASK BULK SOLVENT
Displacement parametersBiso mean: 11.99 Å2
Baniso -1Baniso -2Baniso -3
1-5.326 Å20 Å20 Å2
2---2.249 Å20 Å2
3----3.077 Å2
Refinement stepCycle: LAST / Resolution: 2.79→26.75 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms5716 0 26 45 5787
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.0110.0225868
X-RAY DIFFRACTIONr_angle_refined_deg1.3371.9718018
X-RAY DIFFRACTIONr_dihedral_angle_1_deg5.5785752
X-RAY DIFFRACTIONr_dihedral_angle_2_deg36.54222.661248
X-RAY DIFFRACTIONr_dihedral_angle_3_deg17.30415896
X-RAY DIFFRACTIONr_dihedral_angle_4_deg19.4921560
X-RAY DIFFRACTIONr_chiral_restr0.0910.2934
X-RAY DIFFRACTIONr_gen_planes_refined0.0040.024486
X-RAY DIFFRACTIONr_nbd_refined0.2190.22529
X-RAY DIFFRACTIONr_nbtor_refined0.3140.24029
X-RAY DIFFRACTIONr_xyhbond_nbd_refined0.130.2191
X-RAY DIFFRACTIONr_symmetry_vdw_refined0.1280.220
X-RAY DIFFRACTIONr_symmetry_hbond_refined0.0730.26
X-RAY DIFFRACTIONr_mcbond_it1.4433843
X-RAY DIFFRACTIONr_mcangle_it2.766068
X-RAY DIFFRACTIONr_scbond_it5.453102242
X-RAY DIFFRACTIONr_scangle_it8.431131950
Refine LS restraints NCS

Dom-ID: 1 / Auth asym-ID: A / Refine-ID: X-RAY DIFFRACTION

Ens-IDNumberTypeRms dev position (Å)Weight position
1232TIGHT POSITIONAL0.030.05
1204MEDIUM POSITIONAL0.30.5
1232TIGHT THERMAL0.10.5
1204MEDIUM THERMAL0.882
2540TIGHT POSITIONAL0.050.05
2502MEDIUM POSITIONAL0.460.5
2540TIGHT THERMAL0.10.5
2502MEDIUM THERMAL0.912
3464TIGHT POSITIONAL0.040.05
3425MEDIUM POSITIONAL0.630.5
3464TIGHT THERMAL0.090.5
3425MEDIUM THERMAL0.852
LS refinement shell

Refine-ID: X-RAY DIFFRACTION / Total num. of bins used: 20

Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkNum. reflection all% reflection obs (%)
2.79-2.8610.373880.311595182192.422
2.861-2.9390.332920.2741653179797.106
2.939-3.0240.257830.2151633174498.394
3.024-3.1170.273900.2051581168599.169
3.117-3.2190.3031010.1971528163399.755
3.219-3.3320.223650.1971519158799.811
3.332-3.4580.256960.1861426152399.934
3.458-3.5990.214630.1811435149999.933
3.599-3.7590.221600.181349141099.929
3.759-3.9420.224770.17612981375100
3.942-4.1550.23650.15612271292100
4.155-4.4060.198640.15311631227100
4.406-4.710.174560.15611091165100
4.71-5.0860.218610.15610381099100
5.086-5.570.15510.1669491000100
5.57-6.2250.208390.189863902100
6.225-7.1820.316470.209763810100
7.182-8.7840.262400.168660700100
8.784-12.370.283160.163545561100
12.37-94.9160.36160.28127933488.323
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)
11.0448-0.87850.30614.8879-1.9336.5350.04810.01360.076-0.27940.00510.3019-0.1198-0.7916-0.0532-0.1667-0.02460.0318-0.1268-0.0244-0.1864-28.8901-21.6762-15.4736
21.05920.646-0.1573.0554-0.86792.27050.02090.20730.0017-0.2627-0.0772-0.0402-0.1865-0.04790.0563-0.17610.00330.0363-0.1301-0.0219-0.2065-24.3714-24.0785-24.2159
37.10820.32271.01870.0162-0.044.93640.08760.02920.7375-0.22220.15670.246-0.689-0.4071-0.2443-0.02130.00180.02290.01460.01210.0085-45.6463-44.3384-20.7338
45.4041.5225-0.27486.361-0.65612.8063-0.19780.54820.3649-0.53470.3380.3354-0.4214-0.128-0.1402-0.08340.00230.0235-0.02-0.0642-0.1333-34.7722-43.5582-29.8317
51.49531.7842.34715.97843.53374.18460.14610.3201-0.51170.35160.2765-0.52440.48090.3489-0.42260.02120.06020.1391-0.0293-0.0409-0.1035-19.2546-44.2649-21.4043
61.06120.5578-1.36171.4222-0.74654.3776-0.0495-0.0156-0.04260.1257-0.083-0.12970.1133-0.03720.1326-0.17130.0214-0.0241-0.20520.0004-0.1602-28.8905-44.66212.4666
71.67870.22460.66292.1057-0.43663.9598-0.1306-0.17420.07070.4188-0.0769-0.4289-0.46870.38540.2075-0.1433-0.0447-0.0269-0.1015-0.0482-0.1238-17.9211-29.1088.1777
82.7783-0.32441.83127.17735.739912.94040.1491-0.3733-0.27530.739-0.67451.3069-0.394-1.94740.52540.3410.1541-0.00690.1485-0.05330.1312-32.3556-12.75114.2047
92.52680.1691.00886.64480.24969.11570.0965-0.268-0.32550.79620.06850.36550.0206-0.3375-0.1650.0773-0.0209-0.0835-0.14150.005-0.0896-21.671-19.842620.3608
100.70280.80090.04294.50753.9836.7789-0.15680.08380.0645-0.1440.7723-1.1256-0.61471.0837-0.6155-0.0789-0.0722-0.11050.05350.02010.0154-10.4552-26.67468.5543
Refinement TLS group
IDRefine-IDRefine TLS-IDAuth asym-IDLabel asym-IDAuth seq-IDLabel seq-ID
1X-RAY DIFFRACTION1AA-2 - 6820 - 90
2X-RAY DIFFRACTION2AA69 - 21091 - 232
3X-RAY DIFFRACTION3AA211 - 275233 - 297
4X-RAY DIFFRACTION4AA276 - 338298 - 360
5X-RAY DIFFRACTION5AA339 - 374361 - 396
6X-RAY DIFFRACTION6BB-2 - 16420 - 186
7X-RAY DIFFRACTION7BB165 - 210187 - 232
8X-RAY DIFFRACTION8BB211 - 275233 - 297
9X-RAY DIFFRACTION9BB276 - 338298 - 360
10X-RAY DIFFRACTION10BB339 - 374361 - 396

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