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- PDB-7n7v: Crystal structure of TtnM, a Fe(II)-alpha-ketoglutarate-dependent... -

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Basic information

Entry
Database: PDB / ID: 7n7v
TitleCrystal structure of TtnM, a Fe(II)-alpha-ketoglutarate-dependent hydroxylase from the tautomycetin biosynthesis pathway in Streptomyces griseochromogenes at 2 A.
ComponentsPredicted hydroxylase
KeywordsOXIDOREDUCTASE / Fe(II)-alpha-ketoglutarate-dependent hydroxylase / tautomycetin / Structural Genomics / PSI-Biology / Enzyme Discovery for Natural Product Biosynthesis / NatPro
Function / homologyTauD/TfdA-like domain / Taurine catabolism dioxygenase TauD, TfdA family / Taurine dioxygenase TauD-like superfamily / oxidoreductase activity / : / Predicted hydroxylase
Function and homology information
Biological speciesStreptomyces griseochromogenes (bacteria)
MethodX-RAY DIFFRACTION / SYNCHROTRON / SAD / Resolution: 1.99 Å
AuthorsHan, L. / Xu, W. / Ma, M. / Miller, M.D. / Shen, B. / Phillips Jr., G.N. / Enzyme Discovery for Natural Product Biosynthesis (NatPro)
Funding support United States, 3items
OrganizationGrant numberCountry
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)U01-GM098248 United States
National Institutes of Health/National Cancer Institute (NIH/NCI)R01-CA217255 United States
National Science Foundation (NSF, United States)1231306 United States
CitationJournal: To Be Published
Title: Structure of TtnM, a Fe(II)-alpha-ketoglutarate-dependent hydroxylase from the tautomycetin biosynthesis pathway in Streptomyces griseochromogenes.
Authors: Yang, D. / Yan, W. / Li, G. / Han, L. / Lui, Y.-C. / Ma, M. / Meng, S. / Xie, G. / Xu, W. / Miller, M.D. / Phillips Jr., G.N. / Shen, B.
History
DepositionJun 11, 2021Deposition site: RCSB / Processing site: RCSB
Revision 1.0Jul 6, 2022Provider: repository / Type: Initial release
Revision 1.1May 22, 2024Group: Data collection / Refinement description
Category: chem_comp_atom / chem_comp_bond / struct_ncs_dom_lim
Item: _struct_ncs_dom_lim.beg_auth_comp_id / _struct_ncs_dom_lim.beg_label_asym_id ..._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

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: Predicted hydroxylase
B: Predicted hydroxylase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)83,3487
Polymers83,1292
Non-polymers2185
Water4,558253
1


  • Idetical with deposited unit
  • defined by author
  • Evidence: gel filtration, Gel filtration supports that TtnM runs as a dimer in solution.
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Unit cell
Length a, b, c (Å)50.570, 84.516, 168.920
Angle α, β, γ (deg.)90.000, 90.000, 90.000
Int Tables number19
Space group name H-MP212121
Noncrystallographic symmetry (NCS)NCS domain:
IDEns-IDDetails
11(chain A and (resid 4 through 98 or resid 100...
21(chain B and (resid 4 through 98 or resid 100...

NCS domain segments:

Ens-ID: 1

Dom-IDComponent-IDBeg auth comp-IDBeg label comp-IDEnd auth comp-IDEnd label comp-IDSelection detailsAuth asym-IDLabel asym-IDAuth seq-IDLabel seq-ID
11GLNGLNASNASN(chain A and (resid 4 through 98 or resid 100...AA4 - 9840 - 134
12SERSERARGARG(chain A and (resid 4 through 98 or resid 100...AA100 - 138136 - 174
13ALAALAILEILE(chain A and (resid 4 through 98 or resid 100...AA140 - 208176 - 244
14ARGARGALAALA(chain A and (resid 4 through 98 or resid 100...AA210 - 268246 - 304
15GLYGLYALAALA(chain A and (resid 4 through 98 or resid 100...AA277 - 339313 - 375
16FE2FE2CLCL(chain A and (resid 4 through 98 or resid 100...AC - D401 - 402
21GLNGLNASNASN(chain B and (resid 4 through 98 or resid 100...BB4 - 9840 - 134
22SERSERARGARG(chain B and (resid 4 through 98 or resid 100...BB100 - 138136 - 174
23ALAALAILEILE(chain B and (resid 4 through 98 or resid 100...BB140 - 208176 - 244
24ARGARGALAALA(chain B and (resid 4 through 98 or resid 100...BB210 - 339246 - 375
25FE2FE2CLCL(chain B and (resid 4 through 98 or resid 100...BF - G401 - 402

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Components

#1: Protein Predicted hydroxylase


Mass: 41564.727 Da / Num. of mol.: 2
Source method: isolated from a genetically manipulated source
Details: from cosmid pBS13036
Source: (gene. exp.) Streptomyces griseochromogenes (bacteria)
Gene: ttnM / Plasmid: pBS13039
Details (production host): ttnM gene cloned into HindIII and EcoRI sites of pET28a(+)
Production host: Escherichia coli BL21(DE3) (bacteria) / References: UniProt: C6ZCS6
#2: Chemical ChemComp-FE2 / FE (II) ION


Mass: 55.845 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: Fe / Feature type: SUBJECT OF INVESTIGATION
#3: Chemical ChemComp-CL / CHLORIDE ION


Mass: 35.453 Da / Num. of mol.: 3 / Source method: obtained synthetically / Formula: Cl
#4: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 253 / Source method: isolated from a natural source / Formula: H2O
Has ligand of interestY

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Experimental details

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Experiment

ExperimentMethod: X-RAY DIFFRACTION / Number of used crystals: 2

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Sample preparation

Crystal
IDDensity Matthews3/Da)Density % sol (%)
12.1743.35
2
Crystal growTemperature: 298 K / Method: vapor diffusion, sitting drop / pH: 7.5
Details: 0.2 M magnesium chloride, 0.1 M HEPES, pH 7.5, 25% (w/v) PEG 3350

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Data collection

Diffraction
IDMean temperature (K)Crystal-IDSerial crystal experiment
11001N
21002N
Diffraction source
SourceSiteBeamlineIDWavelength (Å)
SYNCHROTRONAPS 31-ID10.9793
SYNCHROTRONAPS 23-ID-B20.9794
Detector
TypeIDDetectorDate
RAYONIX MX225HE1CCDJul 22, 2015
RAYONIX MX-3002CCDNov 15, 2015
Radiation
IDMonochromatorProtocolMonochromatic (M) / Laue (L)Scattering typeWavelength-ID
1Si(111)SINGLE WAVELENGTHMx-ray1
2Si(111)SINGLE WAVELENGTHMx-ray2
Radiation wavelength
IDWavelength (Å)Relative weight
10.97931
20.97941
ReflectionResolution: 1.99→48.45 Å / Num. obs: 99754 / % possible obs: 99.2 % / Redundancy: 7.029 % / Biso Wilson estimate: 40.06 Å2 / CC1/2: 0.998 / Rmerge(I) obs: 0.101 / Rrim(I) all: 0.109 / Χ2: 0.803 / Net I/σ(I): 11.77
Reflection shell

Diffraction-ID: 1

Resolution (Å)Redundancy (%)Rmerge(I) obsMean I/σ(I) obsNum. unique obsCC1/2Rrim(I) all% possible all
1.99-2.115.1671.5571.176970.5341.73295.6
2.11-2.267.361.0282.3875280.8581.105100
2.26-2.447.4780.6093.8170700.9430.654100
2.44-2.677.4730.3486.165330.9750.374100
2.67-2.997.4780.17210.6459210.9930.185100
2.99-3.457.4410.08419.5752350.9980.09100
3.45-4.227.3360.05529.6944980.9970.06100
4.22-5.967.1810.0535.2735150.9970.055100
5.96-48.456.5510.04436.2420040.9980.04896.8

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Phasing

PhasingMethod: SAD

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Processing

Software
NameVersionClassification
XDSOct 15, 2015data reduction
XDSdata scaling
RESOLVEmodel building
PHENIX1.19.1_4122refinement
PDB_EXTRACT3.27data extraction
AutoSol1.1phasing
RefinementMethod to determine structure: SAD / Resolution: 1.99→48.45 Å / SU ML: 0.29 / Cross valid method: THROUGHOUT / σ(F): 1.91 / Phase error: 27.86 / Stereochemistry target values: ML
RfactorNum. reflection% reflectionSelection details
Rfree0.2233 4277 4.53 %Random
Rwork0.1869 90193 --
obs0.1886 94470 98.85 %-
Solvent computationShrinkage radii: 0.9 Å / VDW probe radii: 1.11 Å / Solvent model: FLAT BULK SOLVENT MODEL
Displacement parametersBiso max: 137.36 Å2 / Biso mean: 55.796 Å2 / Biso min: 22.18 Å2
Refinement stepCycle: final / Resolution: 1.99→48.45 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms4957 0 5 255 5217
Biso mean--46.63 47.52 -
Num. residues----634
Refine LS restraints
Refine-IDTypeDev idealNumber
X-RAY DIFFRACTIONf_bond_d0.00615120
X-RAY DIFFRACTIONf_angle_d0.6236961
X-RAY DIFFRACTIONf_chiral_restr0.0449743
X-RAY DIFFRACTIONf_plane_restr0.0061942
X-RAY DIFFRACTIONf_dihedral_angle_d11.77661935
Refine LS restraints NCS
Ens-IDDom-IDAuth asym-IDNumberRefine-IDTypeRms dev position (Å)
11A3018X-RAY DIFFRACTIONTORSIONAL0.639
12B3018X-RAY DIFFRACTIONTORSIONAL0.639
LS refinement shell

Refine-ID: X-RAY DIFFRACTION / Rfactor Rfree error: 0 / Total num. of bins used: 30

Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkNum. reflection all% reflection obs (%)
1.99-2.020.3849840.40962222230672
2.02-2.040.43161370.401830273164100
2.04-2.060.44021400.361830543194100
2.06-2.090.37641260.374230563182100
2.09-2.120.44481320.322330723204100
2.12-2.150.30531470.317430353182100
2.15-2.180.28141430.295430123155100
2.18-2.210.31041440.285630303174100
2.21-2.240.31061430.26730223165100
2.24-2.280.29211470.269530673214100
2.28-2.320.35981460.25830103156100
2.32-2.360.28711440.238430173161100
2.36-2.410.28731490.238530913240100
2.41-2.460.27421450.218129763121100
2.46-2.510.29871500.230830433193100
2.51-2.570.31411470.227730433190100
2.57-2.630.24711470.214930343181100
2.63-2.70.22151480.211930303178100
2.7-2.780.27121450.204630483193100
2.78-2.870.22551500.190130283178100
2.87-2.980.25641470.187330833230100
2.98-3.10.2171400.187830153155100
3.1-3.240.26451440.183930103154100
3.24-3.410.23361490.177830653214100
3.41-3.620.16461520.163430443196100
3.62-3.90.17891480.145430343182100
3.9-4.290.16621470.134930293176100
4.29-4.910.15371450.126430273172100
4.91-6.180.22541430.141830423185100
6.19-48.450.16631480.16652927307596
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)
13.0840.48572.14972.33820.23954.2066-0.21970.3841-0.5212-0.39940.1135-0.00050.537-0.0125-0.01470.4572-0.06090.05020.3346-0.04780.463510.652-17.753372.6268
22.30030.32170.10572.8454-0.38794.78670.0349-0.0299-0.22240.0096-0.1858-0.10860.3559-0.00890.14010.2320.0203-0.00060.3201-0.06660.334718.2523-7.484680.0097
30.5928-0.0964-0.14834.6813-4.22744.9673-0.05150.14850.0885-0.01510.24170.2521-0.2847-0.6353-0.26440.3380.0237-0.0460.4442-0.09410.3327.66284.139869.4222
42.20410.0041.84222.2251-0.08898.5935-0.20120.22510.19090.1007-0.1498-0.1145-0.91290.44520.33450.558-0.0606-0.04850.34760.01140.383120.346116.611670.1049
54.7232.26692.834.08510.53716.1805-0.2215-0.06270.48320.2159-0.0126-0.0124-1.1256-0.15630.23170.72580.140.0110.3905-0.05790.405913.615522.368172.8522
61.08620.64690.49933.1396-1.12973.4909-0.04970.13750.01780.08430.070.1216-0.2049-0.2803-0.06690.33840.0451-0.00280.3789-0.060.325712.80731.168673.0648
73.1036-1.69462.25716.1409-2.6896.62480.06250.39820.1394-0.1588-0.4264-0.50760.04850.90040.26950.2703-0.0815-0.0380.4968-0.00320.435430.56746.32376.4264
84.9711-1.2544-2.06864.9845-0.27422.7019-0.0295-0.5424-0.51820.0247-0.1221-0.53140.69011.52340.31070.35090.112-0.02310.6209-0.00220.539233.8006-5.81780.2314
90.6822-1.2385-0.52684.6818-0.22450.94330.6375-0.2311-0.90290.46710.3244-0.46371.85830.3504-0.84690.79920.1185-0.0090.38020.0470.437112.4418-14.37925.0269
102.11070.16721.03172.02860.85625.70290.0159-0.0614-0.07760.07860.021-0.08640.59760.031-0.04530.3868-0.00880.04060.30090.04210.29438.1303-2.83823.5275
110.8321-0.21820.01224.5624.42946.5656-0.1685-0.0980.04640.11670.303-0.1619-0.01190.9325-0.22690.40910.041-0.01430.50040.04870.34719.50676.147534.3773
124.62760.20632.27823.1961-1.49516.3621-0.2475-0.05750.50070.0452-0.08010.0584-0.8053-0.22360.29110.53970.052-0.00590.3413-0.01460.33787.331819.951436.6765
135.9417-2.02642.70442.5058-0.95386.1403-0.20870.19730.3197-0.0809-0.11090.0203-1.20630.54310.38510.751-0.1355-0.0340.35710.03750.419614.327225.610334.6275
141.9087-0.0998-0.00882.38611.19747.0384-0.073-0.0379-0.0820.0380.1034-0.19220.15910.4849-0.07810.3743-0.00950.00160.35290.050.347413.60974.574831.469
154.23610.00073.02799.31263.24376.2182-0.0426-0.31540.27130.2024-0.42170.5751-0.5894-0.95640.41580.29590.09840.00210.5238-0.01950.41-3.345511.280428.826
166.2647-1.5352-4.09312.19951.23862.9344-0.16540.2443-0.4142-0.1841-0.22760.93150.5769-1.67340.44250.4318-0.0758-0.00230.6103-0.05130.447-7.2279-0.009423.2894
Refinement TLS group
IDRefine-IDRefine TLS-IDSelection detailsAuth asym-IDAuth seq-ID
1X-RAY DIFFRACTION1chain 'A' and (resid -3 through 27 )A-3 - 27
2X-RAY DIFFRACTION2chain 'A' and ((resid 28 through 131) or (resid 401) )A0
3X-RAY DIFFRACTION3chain 'A' and (resid 132 through 162 )A132 - 162
4X-RAY DIFFRACTION4chain 'A' and (resid 163 through 207 )A163 - 207
5X-RAY DIFFRACTION5chain 'A' and (resid 208 through 239 )A208 - 239
6X-RAY DIFFRACTION6chain 'A' and (resid 240 through 297 )A240 - 297
7X-RAY DIFFRACTION7chain 'A' and (resid 298 through 319 )A298 - 319
8X-RAY DIFFRACTION8chain 'A' and (resid 320 through 339 )A320 - 339
9X-RAY DIFFRACTION9chain 'B' and (resid 4 through 27 )B4 - 27
10X-RAY DIFFRACTION10chain 'B' and ((resid 28 through 131 ) or (resid 401) )B0
11X-RAY DIFFRACTION11chain 'B' and (resid 132 through 162 )B132 - 162
12X-RAY DIFFRACTION12chain 'B' and (resid 163 through 207 )B163 - 207
13X-RAY DIFFRACTION13chain 'B' and (resid 208 through 239 )B208 - 239
14X-RAY DIFFRACTION14chain 'B' and (resid 240 through 297 )B240 - 297
15X-RAY DIFFRACTION15chain 'B' and (resid 298 through 319 )B298 - 319
16X-RAY DIFFRACTION16chain 'B' and (resid 320 through 339 )B320 - 339

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