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- PDB-4xgx: Crystal structure of Escherichia coli Flavin trafficking protein,... -

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Basic information

Entry
Database: PDB / ID: 4xgx
TitleCrystal structure of Escherichia coli Flavin trafficking protein, an FMN transferase, Y60N mutant, ADP-inhibited
ComponentsFAD:protein FMN transferase
KeywordsTRANSFERASE/TRANSFERASE INHIBITOR / flavin transferase / bimetal center / lipoprotein / TRANSFERASE-TRANSFERASE INHIBITOR complex
Function / homology
Function and homology information


FAD:protein FMN transferase / transferase activity / metal ion binding / plasma membrane
Similarity search - Function
T-fold / ApbE-like domains / Flavin transferase ApbE / ApbE family / ApbE-like superfamily / Prokaryotic membrane lipoprotein lipid attachment site profile. / Roll / Alpha Beta
Similarity search - Domain/homology
ADENOSINE-5'-DIPHOSPHATE / NITRATE ION / FAD:protein FMN transferase
Similarity search - Component
Biological speciesEscherichia coli (E. coli)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 1.9 Å
AuthorsTomchick, D.R. / Brautigam, C.A. / Deka, R.K. / Norgard, M.V.
CitationJournal: Microbiologyopen / Year: 2016
Title: Molecular insights into the enzymatic diversity of flavin-trafficking protein (Ftp; formerly ApbE) in flavoprotein biogenesis in the bacterial periplasm.
Authors: Deka, R.K. / Brautigam, C.A. / Liu, W.Z. / Tomchick, D.R. / Norgard, M.V.
History
DepositionJan 2, 2015Deposition site: RCSB / Processing site: RCSB
Revision 1.0Dec 16, 2015Provider: repository / Type: Initial release
Revision 1.1Mar 16, 2016Group: Database references
Revision 1.2Nov 22, 2017Group: Derived calculations / Refinement description / Category: pdbx_struct_oper_list / software
Item: _pdbx_struct_oper_list.symmetry_operation / _software.name
Revision 1.3Sep 27, 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
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_seq_id / _pdbx_struct_conn_angle.ptnr3_auth_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

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: FAD:protein FMN transferase
B: FAD:protein FMN transferase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)76,96121
Polymers75,2032
Non-polymers1,75819
Water6,702372
1


  • Idetical with deposited unit
  • defined by author&software
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area8190 Å2
ΔGint-27 kcal/mol
Surface area23910 Å2
MethodPISA
Unit cell
Length a, b, c (Å)54.583, 56.981, 224.766
Angle α, β, γ (deg.)90.000, 90.000, 90.000
Int Tables number19
Space group name H-MP212121

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Components

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Protein , 1 types, 2 molecules AB

#1: Protein FAD:protein FMN transferase / Flavin transferase


Mass: 37601.367 Da / Num. of mol.: 2 / Fragment: Soluble fragment (UNP residues 21-351) / Mutation: Y60N
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Escherichia coli (E. coli) / Strain: K12 / Gene: apbE, yojK, yojL, b2214, JW5368 / Plasmid: pET-21 NESG / Production host: Escherichia coli (E. coli) / Strain (production host): BL21(DE3) / References: UniProt: P0AB85, FAD:protein FMN transferase

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Non-polymers , 5 types, 391 molecules

#2: Chemical
ChemComp-MG / MAGNESIUM ION


Mass: 24.305 Da / Num. of mol.: 4 / Source method: obtained synthetically / Formula: Mg
#3: Chemical ChemComp-ADP / ADENOSINE-5'-DIPHOSPHATE / Adenosine diphosphate


Mass: 427.201 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: C10H15N5O10P2 / Comment: ADP, energy-carrying molecule*YM
#4: Chemical
ChemComp-NO3 / NITRATE ION / Nitrate


Mass: 62.005 Da / Num. of mol.: 9 / Source method: obtained synthetically / Formula: NO3
#5: Chemical
ChemComp-EDO / 1,2-ETHANEDIOL / ETHYLENE GLYCOL / Ethylene glycol


Mass: 62.068 Da / Num. of mol.: 4 / Source method: obtained synthetically / Formula: C2H6O2
#6: Water ChemComp-HOH / water / Water


Mass: 18.015 Da / Num. of mol.: 372 / 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: 2.32 Å3/Da / Density % sol: 47.03 %
Crystal growTemperature: 293 K / Method: vapor diffusion, hanging drop / pH: 7
Details: 5 mM magnesium chloride, 5 mM ADP, 0.2 M ammonium nitrate, 20% (w/v) PEG 3350, 35% (v/v) ethylene glycol;

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: APS / Beamline: 19-ID / Wavelength: 0.97918 Å
DetectorType: ADSC QUANTUM 315r / Detector: CCD / Date: Mar 4, 2013 / Details: monochromator
RadiationMonochromator: SAGITALLY FOCUSED Si(111) / Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.97918 Å / Relative weight: 1
ReflectionResolution: 1.85→50 Å / Num. obs: 57017 / % possible obs: 93.6 % / Redundancy: 4.1 % / Biso Wilson estimate: 19.09 Å2 / Rmerge(I) obs: 0.039 / Rpim(I) all: 0.02 / Rrim(I) all: 0.044 / Χ2: 0.932 / Net I/av σ(I): 29.6 / Net I/σ(I): 15.2 / Num. measured all: 231169
Reflection shell

Diffraction-ID: 1 / Rejects: 0

Resolution (Å)Redundancy (%)Rmerge(I) obsNum. unique allCC1/2Rpim(I) allRrim(I) allΧ2% possible all
1.85-1.883.40.60618560.7190.3340.6980.91960.7
1.88-1.923.60.51520720.8030.2830.5920.8870.1
1.92-1.953.70.43423170.8520.2390.4990.92276.5
1.95-1.993.90.36724620.9020.20.420.90483.8
1.99-2.043.90.29427680.9370.1610.3370.90191.2
2.04-2.0840.24128920.9520.1330.2770.9296.8
2.08-2.144.10.19929550.970.1090.2280.94198.6
2.14-2.194.10.16530160.9810.090.1890.91599.6
2.19-2.264.20.13829940.9830.0750.1580.89899.9
2.26-2.334.20.10430070.9910.0560.1190.909100
2.33-2.414.20.0930170.9920.0480.1020.921100
2.41-2.514.20.07230210.9940.0390.0820.875100
2.51-2.634.20.05930520.9960.0310.0670.847100
2.63-2.764.20.0530040.9970.0260.0570.80799.9
2.76-2.944.20.0430650.9980.0210.0450.85299.9
2.94-3.164.20.03930640.9980.020.0441.03999.9
3.16-3.484.20.0430330.9970.020.0451.38299.6
3.48-3.994.10.03730930.9970.0180.0411.46699.4
3.99-5.0240.02231020.9990.0120.0250.80598.7
5.02-503.90.01732270.9980.010.020.47597

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Processing

Software
NameVersionClassification
PHENIXrefinement
HKL-3000data reduction
PDB_EXTRACT3.15data extraction
HKL-3000data scaling
PHENIXphasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: PDB ENTRY 4XGV
Resolution: 1.9→28.264 Å / SU ML: 0.16 / Cross valid method: FREE R-VALUE / σ(F): 1.35 / Phase error: 21.09 / Stereochemistry target values: ML
RfactorNum. reflection% reflection
Rfree0.2143 2495 5.01 %
Rwork0.1799 47349 -
obs0.1816 49844 88.44 %
Solvent computationShrinkage radii: 0.9 Å / VDW probe radii: 1.11 Å / Solvent model: FLAT BULK SOLVENT MODEL
Displacement parametersBiso max: 161.91 Å2 / Biso mean: 33.4323 Å2 / Biso min: 3.1 Å2
Refinement stepCycle: final / Resolution: 1.9→28.264 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms4470 0 158 372 5000
Biso mean--42.07 35.05 -
Num. residues----583
Refine LS restraints
Refine-IDTypeDev idealNumber
X-RAY DIFFRACTIONf_bond_d0.0174702
X-RAY DIFFRACTIONf_angle_d0.9856345
X-RAY DIFFRACTIONf_chiral_restr0.035731
X-RAY DIFFRACTIONf_plane_restr0.004806
X-RAY DIFFRACTIONf_dihedral_angle_d14.8641724
LS refinement shell

Refine-ID: X-RAY DIFFRACTION / Total num. of bins used: 18

Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkNum. reflection all% reflection obs (%)
1.9-1.93650.249560.22631110116638
1.9365-1.97610.2131840.22291475155950
1.9761-2.0190.27081070.20411867197464
2.019-2.0660.21731070.212188229575
2.066-2.11760.27371270.19332420254782
2.1176-2.17490.25351360.18422613274990
2.1749-2.23880.24381610.18562805296696
2.2388-2.31110.22831510.17382940309199
2.3111-2.39360.19551340.171929783112100
2.3936-2.48940.20191530.162929503103100
2.4894-2.60260.22321610.16929603121100
2.6026-2.73970.22361540.182629583112100
2.7397-2.91120.211320.177829733105100
2.9112-3.13570.25231420.18930053147100
3.1357-3.45080.22821780.182229713149100
3.4508-3.9490.18581740.17042999317399
3.949-4.97090.17831600.15813024318499
4.9709-28.26750.20891780.1973113329197
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)
10.0072-0.00720.00640.0058-0.00410.00350.0029-0.0004-0.01280.00580.0163-0.03030.00110.02310.00490.1313-0.0827-0.13010.32620.01130.130610.458621.0118-12.5538
20.0001-0.0018-0.00020.0057-0.00080.00070.0043-0.02460.01210.01320.0048-0.004-0.0161-0.00050.00230.23490.00310.01490.1683-0.05280.0583-11.221536.4104-10.7797
30.05080.00810.00010.0289-0.02380.02160.0204-0.1665-0.03350.08620.05430.0776-0.0818-0.1258-0.0070.0506-0.02190.09760.07890.1777-0.0696-18.514526.795-13.0423
40.0078-0.00130.0040.00330.00020.00210.0021-0.0072-0.0038-0.00320.0158-0.0102-0.00220.01320.02790.0733-0.0733-0.0680.23420.05610.08575.954119.5519-20.3139
50.0251-0.02750.02980.0538-0.02530.03250.0164-0.0452-0.03580.02950.07140.06670.012-0.03970.20170.063-0.0455-0.00320.18960.17910.0253-6.47819.9859-14.6374
60.00760.00660.00360.01420.01020.0082-0.0308-0.0164-0.03120.029-0.00970.03360.03990.0001-0.01760.2469-0.09350.00790.13250.11670.239-13.61560.8301-18.0737
70.0045-0.0034-0.00660.0076-0.00030.01510.00510.0173-0.04080.00710.0091-0.02650.01770.00110.01440.15130.03780.0060.0327-0.02310.0865-3.41595.9166-46.0613
80.0045-0.00240.00980.006-0.00770.0259-0.02330.0082-0.056-0.0039-0.01130.00550.0281-0.0119-0.02070.1949-0.0201-0.00410.0431-0.0280.1526-13.4118.3389-43.8036
90.0009-0.00310.0020.01-0.00520.00410.00520.0056-0.0048-0.01570.0030.01550.0005-0.0140.00610.10450.0646-0.05050.12050.04650.0855-21.122730.2843-43.984
100.0031-0.0050.00230.0192-0.00890.0043-0.01350.0089-0.0057-0.0345-0.0044-0.002-0.03380.0048-0.0110.0792-0.00770.04860.01640.06020.1204-6.504833.8465-40.5426
110.00160.0023-0.00520.0038-0.00770.01570.00550.0074-0.006-0.02350.01150.00390.0046-0.0030.01630.25210.01230.07250.07920.02810.1336-6.566444.1617-55.9428
120.0115-0.0167-0.0060.03310.00250.00710.01790.0057-0.0043-0.03050.00190.0042-0.0259-0.02670.02670.08960.09460.06710.04960.06990.1365-15.81134.1564-36.6651
130.007900.00370.0160.00240.00530.0221-0.0108-0.06410.0148-0.0265-0.02930.02060.0124-0.00380.1660.03190.00610.02290.02050.1233-1.07289.291-36.889
140.02270.0024-0.00120.00080.00240.01780.0143-0.0043-0.0029-0.0280.0101-0.0099-0.0090.00860.04340.11460.03230.07410.07190.04110.07765.423120.5815-44.6481
150.01140.0188-0.01590.0295-0.02550.02210.00890.0420.0347-0.0445-0.0058-0.03-0.03460.0182-0.01550.06920.00690.10150.07280.08630.1074.288531.2472-43.3465
160.00930.0031-0.00260.0047-0.00170.001-0.00230.02280.0234-0.0163-0.0121-0.0206-0.00980.0107-0.00380.1063-0.0830.03350.23340.0720.25912.764132.2493-39.2291
170.0267-0.0022-0.00920.0065-0.0050.00880.01140.0093-0.013-0.00480.001-0.0069-0.00170.01570.0020.076-0.00980.03630.28870.06290.271517.65823.1926-40.0127
Refinement TLS group
IDRefine-IDRefine TLS-IDSelection detailsAuth asym-IDAuth seq-ID
1X-RAY DIFFRACTION1chain 'A' and (resid 13 through 57 )A0
2X-RAY DIFFRACTION2chain 'A' and (resid 58 through 81 )A0
3X-RAY DIFFRACTION3chain 'A' and (resid 82 through 167 )A0
4X-RAY DIFFRACTION4chain 'A' and (resid 168 through 192 )A0
5X-RAY DIFFRACTION5chain 'A' and (resid 193 through 289 )A0
6X-RAY DIFFRACTION6chain 'A' and (resid 290 through 329 )A0
7X-RAY DIFFRACTION7chain 'B' and (resid 13 through 35 )B0
8X-RAY DIFFRACTION8chain 'B' and (resid 36 through 57 )B0
9X-RAY DIFFRACTION9chain 'B' and (resid 58 through 81 )B0
10X-RAY DIFFRACTION10chain 'B' and (resid 82 through 113 )B0
11X-RAY DIFFRACTION11chain 'B' and (resid 114 through 136 )B0
12X-RAY DIFFRACTION12chain 'B' and (resid 137 through 167 )B0
13X-RAY DIFFRACTION13chain 'B' and (resid 168 through 209 )B0
14X-RAY DIFFRACTION14chain 'B' and (resid 210 through 255 )B0
15X-RAY DIFFRACTION15chain 'B' and (resid 256 through 289 )B0
16X-RAY DIFFRACTION16chain 'B' and (resid 290 through 311 )B0
17X-RAY DIFFRACTION17chain 'B' and (resid 312 through 329 )B0

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