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- PDB-3c1q: The three-dimensional structure of the cytoplasmic domains of Eps... -

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Basic information

Entry
Database: PDB / ID: 3c1q
TitleThe three-dimensional structure of the cytoplasmic domains of EpsF from the Type 2 Secretion System of Vibrio cholerae
ComponentsGeneral secretion pathway protein F
KeywordsTRANSPORT PROTEIN / Type 2 Secretion System / T2SS / T4PB / Cholera / Inner membrane / Membrane / Transmembrane
Function / homology
Function and homology information


protein secretion by the type II secretion system / type II protein secretion system complex / metal ion binding / plasma membrane
Similarity search - Function
Type II secretion system protein GspF / Type II secretion system (T2SS), domain F / T2SS_GspF/T4SS_PilC conserved site / Bacterial type II secretion system protein F signature. / GspF/PilC family / Type II secretion system protein GspF domain / Type II secretion system GspF domain superfamily / Type II secretion system (T2SS), protein F / Receptor-associated Protein / Up-down Bundle / Mainly Alpha
Similarity search - Domain/homology
Type II secretion system protein F
Similarity search - Component
Biological speciesVibrio cholerae (bacteria)
MethodX-RAY DIFFRACTION / MOLECULAR REPLACEMENT / Resolution: 1.7 Å
AuthorsAbendroth, J. / Mitchell, D.D. / Korotkov, K.V. / Kreeger, A. / Hol, W.G.J.
CitationJournal: J.Struct.Biol. / Year: 2009
Title: The three-dimensional structure of the cytoplasmic domains of EpsF from the type 2 secretion system of Vibrio cholerae
Authors: Abendroth, J. / Mitchell, D.D. / Korotkov, K.V. / Johnson, T.L. / Kreger, A. / Sandkvist, M. / Hol, W.G.
History
DepositionJan 24, 2008Deposition site: RCSB / Processing site: RCSB
Revision 1.0Feb 3, 2009Provider: repository / Type: Initial release
Revision 1.1Jul 13, 2011Group: Advisory / Version format compliance
Revision 1.2Aug 30, 2023Group: Advisory / Data collection ...Advisory / Data collection / Database references / Derived calculations / Refinement description / Structure summary
Category: chem_comp / chem_comp_atom ...chem_comp / chem_comp_atom / chem_comp_bond / database_2 / pdbx_initial_refinement_model / pdbx_struct_conn_angle / pdbx_unobs_or_zero_occ_atoms / struct_conn / struct_conn_type / struct_ncs_dom_lim / struct_ref_seq_dif / struct_site
Item: _chem_comp.pdbx_synonyms / _database_2.pdbx_DOI ..._chem_comp.pdbx_synonyms / _database_2.pdbx_DOI / _database_2.pdbx_database_accession / _pdbx_struct_conn_angle.ptnr1_auth_asym_id / _pdbx_struct_conn_angle.ptnr1_auth_comp_id / _pdbx_struct_conn_angle.ptnr1_auth_seq_id / _pdbx_struct_conn_angle.ptnr1_label_asym_id / _pdbx_struct_conn_angle.ptnr1_label_atom_id / _pdbx_struct_conn_angle.ptnr1_label_comp_id / _pdbx_struct_conn_angle.ptnr1_label_seq_id / _pdbx_struct_conn_angle.ptnr2_auth_asym_id / _pdbx_struct_conn_angle.ptnr2_auth_comp_id / _pdbx_struct_conn_angle.ptnr2_auth_seq_id / _pdbx_struct_conn_angle.ptnr2_label_asym_id / _pdbx_struct_conn_angle.ptnr2_label_atom_id / _pdbx_struct_conn_angle.ptnr2_label_comp_id / _pdbx_struct_conn_angle.ptnr3_auth_asym_id / _pdbx_struct_conn_angle.ptnr3_auth_comp_id / _pdbx_struct_conn_angle.ptnr3_auth_seq_id / _pdbx_struct_conn_angle.ptnr3_label_asym_id / _pdbx_struct_conn_angle.ptnr3_label_atom_id / _pdbx_struct_conn_angle.ptnr3_label_comp_id / _pdbx_struct_conn_angle.ptnr3_label_seq_id / _pdbx_struct_conn_angle.value / _struct_conn.conn_type_id / _struct_conn.id / _struct_conn.pdbx_dist_value / _struct_conn.pdbx_leaving_atom_flag / _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 / _struct_conn.ptnr2_label_seq_id / _struct_conn_type.id / _struct_ncs_dom_lim.beg_auth_comp_id / _struct_ncs_dom_lim.beg_label_asym_id / _struct_ncs_dom_lim.beg_label_seq_id / _struct_ncs_dom_lim.end_auth_comp_id / _struct_ref_seq_dif.details / _struct_site.pdbx_auth_asym_id / _struct_site.pdbx_auth_comp_id / _struct_site.pdbx_auth_seq_id
Revision 1.3Nov 15, 2023Group: Data collection / Category: chem_comp_atom / chem_comp_bond / Item: _chem_comp_atom.atom_id / _chem_comp_bond.atom_id_2

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: General secretion pathway protein F
B: General secretion pathway protein F
hetero molecules


Theoretical massNumber of molelcules
Total (without water)28,6007
Polymers28,0632
Non-polymers5375
Water6,035335
1


  • Idetical with deposited unit
  • defined by author&software
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area2850 Å2
MethodPISA
Unit cell
Length a, b, c (Å)46.988, 52.122, 89.470
Angle α, β, γ (deg.)90.00, 90.00, 90.00
Int Tables number19
Space group name H-MP212121
Noncrystallographic symmetry (NCS)NCS domain:
IDEns-IDDetails
11A
21B

NCS domain segments:

Component-ID: 1 / Ens-ID: 1 / End auth comp-ID: TYR / End label comp-ID: TYR / End label seq-ID: 71 / Refine code: 6 / Auth seq-ID: 1 - 125

Dom-IDBeg auth comp-IDBeg label comp-IDAuth asym-IDLabel asym-ID
1CLCLAD - A
2CACABG - B

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Components

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Protein , 1 types, 2 molecules AB

#1: Protein General secretion pathway protein F / Cholera toxin secretion protein epsF


Mass: 14031.403 Da / Num. of mol.: 2 / Fragment: cyto1-EpsF (UNP residues 56-170)
Source method: isolated from a genetically manipulated source
Details: C-terminal TEV-cleavable His-tag / Source: (gene. exp.) Vibrio cholerae (bacteria) / Gene: epsF / Plasmid: pACYC-CT / Production host: Escherichia coli (E. coli) / Strain (production host): BL21(DE3) / References: UniProt: P45780

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Non-polymers , 5 types, 340 molecules

#2: Chemical ChemComp-CA / CALCIUM ION


Mass: 40.078 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: Ca
#3: Chemical ChemComp-CL / CHLORIDE ION


Mass: 35.453 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: Cl
#4: Chemical ChemComp-NA / SODIUM ION


Mass: 22.990 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: Na
#5: Chemical ChemComp-PE5 / 3,6,9,12,15,18,21,24-OCTAOXAHEXACOSAN-1-OL / 2-(2-{2-[2-(2-{2-[2-(2-ETHOXY-ETHOXY)-ETHOXY]-ETHOXY}-ETHOXY)-ETHOXY]-ETHOXY}-ETHOXY)-ETHANOL / POLYETHYLENE GLYCOL PEG400


Mass: 398.489 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: C18H38O9 / Comment: precipitant*YM
#6: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 335 / 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: 1.95 Å3/Da / Density % sol: 36.99 %
Crystal growTemperature: 293 K / Method: vapor diffusion, sitting drop / pH: 7
Details: 1ul protein at 10mg/ml + 1ul reservoir: 12.5% PEG 400, 200mM CaOAc2, 100mM MES pH 7.0, VAPOR DIFFUSION, SITTING DROP, temperature 293K

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: ROTATING ANODE / Type: RIGAKU MICROMAX-007 HF
DetectorType: RIGAKU SATURN 944 / Detector: CCD / Date: Oct 30, 2006 / Details: multilayer (Varimax)
RadiationMonochromator: multilayer / Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthRelative weight: 1
ReflectionResolution: 1.7→45.05 Å / Num. all: 23838 / Num. obs: 23838 / % possible obs: 95.6 % / Redundancy: 3.78 % / Biso Wilson estimate: 17.5 Å2 / Rmerge(I) obs: 0.06 / Rsym value: 0.06 / Net I/σ(I): 15
Reflection shellResolution: 1.7→1.76 Å / Redundancy: 1.97 % / Rmerge(I) obs: 0.223 / Mean I/σ(I) obs: 3.3 / Num. unique all: 1347 / Rsym value: 0.223 / % possible all: 79.1

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Processing

Software
NameVersionClassification
REFMAC5.3.0021refinement
CrystalCleardata collection
CrystalCleardata reduction
CrystalCleardata scaling
MOLREPphasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: 2vma, 2vmb
Resolution: 1.7→33.94 Å / Cor.coef. Fo:Fc: 0.963 / Cor.coef. Fo:Fc free: 0.94 / SU B: 3.737 / SU ML: 0.067 / TLS residual ADP flag: LIKELY RESIDUAL / Isotropic thermal model: isotropic with TLS / Cross valid method: THROUGHOUT / σ(F): 0 / σ(I): 0 / ESU R: 0.109 / ESU R Free: 0.114 / Stereochemistry target values: MAXIMUM LIKELIHOOD / Details: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS
RfactorNum. reflection% reflectionSelection details
Rfree0.20793 1223 5.1 %RANDOM
Rwork0.15587 ---
all0.15846 22632 --
obs0.15846 22632 95.96 %-
Solvent computationIon probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.4 Å / Solvent model: MASK
Displacement parametersBiso mean: 9.193 Å2
Baniso -1Baniso -2Baniso -3
1-0 Å20 Å20 Å2
2--0.48 Å20 Å2
3----0.48 Å2
Refinement stepCycle: LAST / Resolution: 1.7→33.94 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms1808 0 20 335 2163
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.0120.0221888
X-RAY DIFFRACTIONr_bond_other_d0.0020.021332
X-RAY DIFFRACTIONr_angle_refined_deg1.27422544
X-RAY DIFFRACTIONr_angle_other_deg0.9463.0023220
X-RAY DIFFRACTIONr_dihedral_angle_1_deg3.9255243
X-RAY DIFFRACTIONr_dihedral_angle_2_deg29.16723.48886
X-RAY DIFFRACTIONr_dihedral_angle_3_deg10.96415356
X-RAY DIFFRACTIONr_dihedral_angle_4_deg15.6221519
X-RAY DIFFRACTIONr_chiral_restr0.0720.2289
X-RAY DIFFRACTIONr_gen_planes_refined0.0050.022084
X-RAY DIFFRACTIONr_gen_planes_other0.0010.02375
X-RAY DIFFRACTIONr_nbd_refined0.2460.2494
X-RAY DIFFRACTIONr_nbd_other0.1970.21407
X-RAY DIFFRACTIONr_nbtor_refined0.1730.2966
X-RAY DIFFRACTIONr_nbtor_other0.0820.2958
X-RAY DIFFRACTIONr_xyhbond_nbd_refined0.1810.2227
X-RAY DIFFRACTIONr_xyhbond_nbd_other
X-RAY DIFFRACTIONr_metal_ion_refined0.1090.29
X-RAY DIFFRACTIONr_metal_ion_other
X-RAY DIFFRACTIONr_symmetry_vdw_refined0.2350.220
X-RAY DIFFRACTIONr_symmetry_vdw_other0.270.274
X-RAY DIFFRACTIONr_symmetry_hbond_refined0.2410.243
X-RAY DIFFRACTIONr_symmetry_hbond_other
X-RAY DIFFRACTIONr_symmetry_metal_ion_refined0.0890.21
X-RAY DIFFRACTIONr_symmetry_metal_ion_other
X-RAY DIFFRACTIONr_mcbond_it0.5191.51171
X-RAY DIFFRACTIONr_mcbond_other0.1631.5471
X-RAY DIFFRACTIONr_mcangle_it0.91921885
X-RAY DIFFRACTIONr_scbond_it1.9983724
X-RAY DIFFRACTIONr_scangle_it3.1784.5652
X-RAY DIFFRACTIONr_rigid_bond_restr
X-RAY DIFFRACTIONr_sphericity_free
X-RAY DIFFRACTIONr_sphericity_bonded
Refine LS restraints NCS

Ens-ID: 1 / Number: 803 / Refine-ID: X-RAY DIFFRACTION

Dom-IDAuth asym-IDTypeRms dev position (Å)Weight position
1Aloose positional0.575
2Bloose thermal1.610
LS refinement shellResolution: 1.7→1.744 Å / Total num. of bins used: 20
RfactorNum. reflection% reflection
Rfree0.303 82 -
Rwork0.189 1347 -
obs-1347 78.91 %
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)
10.9467-0.1106-0.020.6279-0.29671.357-0.00050.08970.0053-0.06230.03330.02190.04080.0093-0.03270.04970.007-0.00750.0580.00550.042322.258524.390545.3776
23.7566-0.45971.15370.0711-0.04433.5867-0.1329-0.0341-0.1040.02610.03050.08270.0316-0.17890.10250.0519-0.0102-0.00010.0211-0.01740.054515.968718.411149.0719
34.2986-0.84311.2521.3752-1.63836.7424-0.21520.1318-0.4054-0.07560.1049-0.02710.14130.1470.11020.0538-0.03260.0462-0.0062-0.02770.071822.117511.368146.4727
42.68810.80850.04831.1932-0.26241.4821-0.07110.0723-0.0543-0.0660.05630.00050.08720.13880.01480.0320.00540.00850.0286-0.00310.046231.904618.718444.9434
52.1991-1.96890.30614.8848-2.18873.2394-0.05050.00760.14660.0935-0.0556-0.17060.03530.19890.10610.0287-0.0080.01170.0375-0.00280.06929.332529.588851.149
62.709-1.47241.06529.4873-5.83358.5568-0.03910.1414-0.2282-0.12510.0974-0.08130.2335-0.1393-0.05830.0515-0.00140.03730.0155-0.01060.016922.351242.843238.0198
71.62350.04460.11275.1366-2.23632.3857-0.01960.00310.06610.13120.01980.0224-0.09080.0338-0.00020.04210.00490.00280.0444-0.00390.027514.879730.894459.0523
84.10691.4746-1.10355.55641.3322.5694-0.0103-0.23880.02020.12980.1339-0.2386-0.09160.1965-0.12360.05640.0115-0.03680.0901-0.01770.009421.725632.063965.6565
94.27673.7975-2.40086.3994-1.82645.3590.0173-0.1669-0.16780.255-0.0164-0.1462-0.0010.0455-0.00090.04380.0096-0.03430.077-0.02540.012316.86633.843772.0429
101.82320.32880.71551.31610.92322.1773-0.0214-0.1430.1313-0.0298-0.01960.1408-0.0913-0.27020.04110.03790.00230.00380.0504-0.00640.02485.125634.796265.9882
113.72863.5884-1.70716.8405-3.55514.26430.04660.05780.1488-0.0740.05220.1038-0.1518-0.0687-0.09880.03750.0083-0.01130.01850.01320.04939.027134.917552.2777
123.17142.971-2.43658.8641-2.68675.1628-0.0927-0.0832-0.0315-0.19530.0918-0.10920.08540.11950.00090.00730.01470.00840.05230.01160.043517.425849.44841.6722
Refinement TLS group
IDRefine-IDRefine TLS-IDAuth asym-IDLabel asym-IDAuth seq-IDLabel seq-ID
1X-RAY DIFFRACTION1AA57 - 833 - 29
2X-RAY DIFFRACTION2AA84 - 10230 - 48
3X-RAY DIFFRACTION3AA103 - 11549 - 61
4X-RAY DIFFRACTION4AA116 - 14462 - 90
5X-RAY DIFFRACTION5AA145 - 16191 - 107
6X-RAY DIFFRACTION6AA162 - 171108 - 117
7X-RAY DIFFRACTION7BB63 - 859 - 31
8X-RAY DIFFRACTION8BB86 - 10032 - 46
9X-RAY DIFFRACTION9BB101 - 11447 - 60
10X-RAY DIFFRACTION10BB115 - 14961 - 95
11X-RAY DIFFRACTION11BB150 - 16196 - 107
12X-RAY DIFFRACTION12BB162 - 177108 - 123

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