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- PDB-6fp8: mTFP1/DARPin 1238_E11 complex in space group C2 -

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Basic information

Entry
Database: PDB / ID: 6fp8
TitlemTFP1/DARPin 1238_E11 complex in space group C2
Components
  • DARPin1238_E11
  • GFP-like fluorescent chromoprotein cFP484
KeywordsDE NOVO PROTEIN / Darpin / protein binder / designed ankyrin repeat protein
Function / homology
Function and homology information


bioluminescence / generation of precursor metabolites and energy
Similarity search - Function
Ankyrin repeat-containing domain / Green fluorescent protein, GFP / Green fluorescent protein-related / Green fluorescent protein / Green fluorescent protein / Serine Threonine Protein Phosphatase 5, Tetratricopeptide repeat / Alpha Horseshoe / Mainly Alpha
Similarity search - Domain/homology
GFP-like fluorescent chromoprotein cFP484
Similarity search - Component
Biological speciesClavularia sp. (invertebrata)
synthetic construct (others)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 1.855 Å
AuthorsJakob, R.P. / Vigano, M.A. / Bieli, D. / Matsuda, S. / Schaefer, J.V. / Pluckthun, A. / Affolter, M. / Maier, T.
CitationJournal: Biol Open / Year: 2018
Title: DARPins recognizing mTFP1 as novel reagents forin vitroandin vivoprotein manipulations.
Authors: Vigano, M.A. / Bieli, D. / Schaefer, J.V. / Jakob, R.P. / Matsuda, S. / Maier, T. / Pluckthun, A. / Affolter, M.
History
DepositionFeb 9, 2018Deposition site: PDBE / Processing site: PDBE
Revision 1.0Oct 3, 2018Provider: repository / Type: Initial release
Revision 1.1Nov 7, 2018Group: Data collection / Database references / Category: citation / citation_author
Item: _citation.journal_volume / _citation.title / _citation_author.identifier_ORCID
Revision 1.2Dec 26, 2018Group: Data collection / Database references / Category: citation / Item: _citation.title
Revision 2.0Sep 1, 2021Group: Database references / Derived calculations ...Database references / Derived calculations / Non-polymer description / Source and taxonomy / Structure summary
Category: chem_comp / database_2 ...chem_comp / database_2 / entity / entity_src_gen / struct_conn / struct_ref / struct_ref_seq / struct_ref_seq_dif
Item: _chem_comp.formula / _chem_comp.formula_weight ..._chem_comp.formula / _chem_comp.formula_weight / _chem_comp.id / _chem_comp.mon_nstd_flag / _chem_comp.name / _chem_comp.pdbx_synonyms / _chem_comp.type / _database_2.pdbx_DOI / _database_2.pdbx_database_accession / _entity.pdbx_description / _entity_src_gen.gene_src_common_name / _entity_src_gen.pdbx_gene_src_ncbi_taxonomy_id / _entity_src_gen.pdbx_gene_src_scientific_name / _struct_conn.pdbx_leaving_atom_flag / _struct_ref.db_code / _struct_ref.db_name / _struct_ref.pdbx_align_begin / _struct_ref.pdbx_db_accession / _struct_ref.pdbx_seq_one_letter_code / _struct_ref_seq.db_align_beg / _struct_ref_seq.db_align_end / _struct_ref_seq.pdbx_auth_seq_align_beg / _struct_ref_seq.pdbx_auth_seq_align_end / _struct_ref_seq.pdbx_db_accession / _struct_ref_seq.seq_align_beg / _struct_ref_seq.seq_align_end
Revision 3.0Nov 15, 2023Group: Atomic model / Data collection
Category: atom_site / atom_site_anisotrop ...atom_site / atom_site_anisotrop / chem_comp_atom / chem_comp_bond
Item: _atom_site.auth_atom_id / _atom_site.label_atom_id ..._atom_site.auth_atom_id / _atom_site.label_atom_id / _atom_site_anisotrop.pdbx_auth_atom_id / _atom_site_anisotrop.pdbx_label_atom_id
Revision 3.1Jan 17, 2024Group: Refinement description / Category: pdbx_initial_refinement_model

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: GFP-like fluorescent chromoprotein cFP484
B: DARPin1238_E11
hetero molecules


Theoretical massNumber of molelcules
Total (without water)49,28811
Polymers48,4262
Non-polymers8639
Water7,386410
1


  • Idetical with deposited unit
  • defined by author&software
  • Evidence: gel filtration
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area4390 Å2
ΔGint-83 kcal/mol
Surface area16620 Å2
MethodPISA
Unit cell
Length a, b, c (Å)135.953, 75.012, 48.295
Angle α, β, γ (deg.)90.00, 90.73, 90.00
Int Tables number5
Space group name H-MC121

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Components

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Protein , 2 types, 2 molecules AB

#1: Protein GFP-like fluorescent chromoprotein cFP484


Mass: 28546.164 Da / Num. of mol.: 1
Source method: isolated from a genetically manipulated source
Details: mTFP1, contains cofactor PIA, made out of amino acids AYG
Source: (gene. exp.) Clavularia sp. (invertebrata) / Plasmid: pRSFDUET-1 / Production host: Escherichia coli BL21(DE3) (bacteria) / References: UniProt: Q9U6Y3
#2: Protein DARPin1238_E11


Mass: 19879.373 Da / Num. of mol.: 1
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) synthetic construct (others) / Plasmid: pQiq / Production host: Escherichia coli (E. coli) / Strain (production host): XL1Blue

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Non-polymers , 4 types, 419 molecules

#3: Chemical ChemComp-MPD / (4S)-2-METHYL-2,4-PENTANEDIOL


Mass: 118.174 Da / Num. of mol.: 3 / Source method: obtained synthetically / Formula: C6H14O2 / Comment: precipitant*YM
#4: Chemical
ChemComp-SO4 / SULFATE ION


Mass: 96.063 Da / Num. of mol.: 4 / Source method: obtained synthetically / Formula: SO4
#5: Chemical ChemComp-EDO / 1,2-ETHANEDIOL / ETHYLENE GLYCOL


Mass: 62.068 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: C2H6O2
#6: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 410 / 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: 57.04 %
Crystal growTemperature: 277 K / Method: vapor diffusion, sitting drop / pH: 7.4
Details: 10% PEG4000, 20% Glycerol, and 0.02 M of L-glutamate, glycine, DL-alanine, L-lysin, DL-serin

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: SLS / Beamline: X06SA / Wavelength: 0.99986 Å
DetectorType: DECTRIS EIGER X 16M / Detector: PIXEL / Date: Nov 19, 2016
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.99986 Å / Relative weight: 1
ReflectionResolution: 1.855→39.607 Å / Num. obs: 78111 / % possible obs: 96.6 % / Redundancy: 2.4 % / CC1/2: 0.998 / Rrim(I) all: 0.069 / Net I/σ(I): 11.2
Reflection shellResolution: 1.855→1.97 Å / Redundancy: 2.3 % / Mean I/σ(I) obs: 1.7 / Num. unique obs: 12129 / CC1/2: 0.652 / Rrim(I) all: 0.73 / % possible all: 93.1

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Processing

Software
NameVersionClassification
PHENIX(1.10pre_2131: ???)refinement
XDSdata reduction
XSCALEdata scaling
PHASERphasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: 2HQK
Resolution: 1.855→39.607 Å / SU ML: 0.19 / Cross valid method: FREE R-VALUE / σ(F): 1.21 / Phase error: 18.14
RfactorNum. reflection% reflection
Rfree0.1764 3897 4.99 %
Rwork0.1483 --
obs0.1498 78111 96.62 %
Solvent computationShrinkage radii: 0.9 Å / VDW probe radii: 1.11 Å
Refinement stepCycle: LAST / Resolution: 1.855→39.607 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms3029 0 52 411 3492
Refine LS restraints
Refine-IDTypeDev idealNumber
X-RAY DIFFRACTIONf_bond_d0.0123164
X-RAY DIFFRACTIONf_angle_d1.3014284
X-RAY DIFFRACTIONf_dihedral_angle_d12.1631877
X-RAY DIFFRACTIONf_chiral_restr0.066462
X-RAY DIFFRACTIONf_plane_restr0.008564
LS refinement shell
Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkRefine-ID% reflection obs (%)
1.8546-1.87720.31191090.30092142X-RAY DIFFRACTION79
1.8772-1.9010.29751420.26122677X-RAY DIFFRACTION97
1.901-1.9260.27871410.25172651X-RAY DIFFRACTION97
1.926-1.95240.2651360.24742635X-RAY DIFFRACTION97
1.9524-1.98020.22981410.24242686X-RAY DIFFRACTION97
1.9802-2.00980.2311440.2252679X-RAY DIFFRACTION97
2.0098-2.04120.23581400.20562652X-RAY DIFFRACTION97
2.0412-2.07470.22971350.19772680X-RAY DIFFRACTION97
2.0747-2.11040.18971390.19352693X-RAY DIFFRACTION98
2.1104-2.14880.21531430.17392678X-RAY DIFFRACTION97
2.1488-2.19010.20411390.17492667X-RAY DIFFRACTION97
2.1901-2.23480.21131420.17382688X-RAY DIFFRACTION99
2.2348-2.28340.22541450.16492701X-RAY DIFFRACTION97
2.2834-2.33650.20141370.1552674X-RAY DIFFRACTION98
2.3365-2.3950.1861400.15192677X-RAY DIFFRACTION98
2.395-2.45970.16911400.14522652X-RAY DIFFRACTION98
2.4597-2.53210.1581410.15232699X-RAY DIFFRACTION97
2.5321-2.61380.20751310.16232509X-RAY DIFFRACTION92
2.6138-2.70720.16761390.14192661X-RAY DIFFRACTION96
2.7072-2.81560.15961410.13892671X-RAY DIFFRACTION99
2.8156-2.94370.19221420.14092720X-RAY DIFFRACTION99
2.9437-3.09880.17511450.14582703X-RAY DIFFRACTION99
3.0988-3.29290.15071400.13022704X-RAY DIFFRACTION98
3.2929-3.54690.14861440.12482713X-RAY DIFFRACTION98
3.5469-3.90360.13461440.11062698X-RAY DIFFRACTION98
3.9036-4.46780.15661380.11132652X-RAY DIFFRACTION97
4.4678-5.62640.14851330.12372562X-RAY DIFFRACTION93
5.6264-39.6160.18471460.16642690X-RAY DIFFRACTION98
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)
10.68240.1242-0.13440.30170.19610.18950.00270.0029-0.13710.00430.0469-0.01670.10450.046400.16010.01410.00230.13750.01760.171-12.7185-2.774120.1393
20.3729-0.16410.08680.18590.1940.5118-0.0282-0.0253-0.0153-0.01290.0169-0.0262-0.03010.0278-00.15610.01430.01280.15340.02330.146-15.90054.675621.5044
30.0490.03650.02860.05640.06640.0248-0.1039-0.0049-0.0632-0.02380.11690.0787-0.0033-0.172100.20870.01650.00080.19440.03650.2003-21.7035-0.113618.906
40.0667-0.0485-0.11840.0143-0.02790.0390.0427-0.0133-0.1037-0.08630.05480.08960.1186-0.113100.2048-0.0057-0.01390.16380.03190.1895-18.1996-2.730218.2881
50.01710.0029-0.00190.04210.04710.04540.30810.35390.1167-0.423-0.2762-0.0843-0.32720.233100.36170.01920.03560.27530.09810.2221-12.221215.80512.9261
60.0339-0.1924-0.00250.11250.03430.0862-0.06570.03060.05970.05640.0896-0.0492-0.1684-0.040100.21110.02290.00020.21070.01730.1992-19.150514.616824.9397
70.0546-0.09950.06190.082-0.05060.0070.0144-0.1158-0.15020.0083-0.1278-0.0945-0.2048-0.0112-00.25650.02650.0290.1704-0.00550.1924-18.140216.204621.5147
80.4807-0.10590.23790.22550.28980.3624-0.0205-0.06080.078-0.0857-0.0090.1215-0.0913-0.1311-00.1830.0360.00850.16940.02320.1362-21.613411.681724.4881
90.2383-0.07520.27190.1906-0.02140.10260.05990.11120.0988-0.0295-0.0263-0.1394-0.1566-0.045-00.17290.00620.01610.1740.01150.181-6.44668.360518.9961
100.60710.41390.35310.3854-0.45610.2890.1084-0.18460.2773-0.05990.1184-0.1522-0.31540.026500.19010.0257-0.00660.20480.01640.1802-10.63819.885428.9323
110.0084-0.0594-0.1291-0.00790.10360.1333-0.2731-0.57740.21580.9741-0.10790.564-0.0834-0.023400.34690.00090.21430.3848-0.00950.4291-45.25971.7121.7785
12-0.04520.1626-0.0780.05050.20590.00590.0766-0.09710.09090.0283-0.11090.09740.0026-0.078700.2143-0.0147-0.01010.25890.00160.2901-40.6455-2.34715.4343
130.09170.01430.03990.03350.15230.10070.03650.0649-0.0398-0.0682-0.0290.00390.0217-0.029900.1723-0.0152-0.04220.22120.01020.2714-34.0769-4.78528.9891
140.0107-0.0049-0.0111-0.0128-0.00450.0035-0.19110.1505-0.1525-0.0003-0.28920.00430.1477-0.224500.2335-0.0806-0.00570.2871-0.0350.2982-39.9658-15.28338.9831
150.1506-0.0283-0.12260.10730.14610.24970.00330.073-0.054-0.02950.00240.04190.0144-0.032300.2133-0.0299-0.04150.2201-0.01720.2549-28.9565-13.29664.4457
160.0406-0.01110.01480.15570.05510.1063-0.02610.1717-0.1634-0.19090.0324-0.1310.0240.003600.2454-0.0112-0.02460.2295-0.05730.271-21.0626-14.39810.3405
170.02680.06470.00040.03730.00360.0175-0.23120.2038-0.20230.1572-0.2163-0.21350.21930.0443-00.4175-0.05040.01030.2503-0.07960.4687-25.0232-24.7265-4.9844
Refinement TLS group
IDRefine-IDRefine TLS-IDSelection details
1X-RAY DIFFRACTION1chain 'A' and (resid 2 through 35 )
2X-RAY DIFFRACTION2chain 'A' and (resid 36 through 99 )
3X-RAY DIFFRACTION3chain 'A' and (resid 100 through 110 )
4X-RAY DIFFRACTION4chain 'A' and (resid 111 through 123 )
5X-RAY DIFFRACTION5chain 'A' and (resid 124 through 135 )
6X-RAY DIFFRACTION6chain 'A' and (resid 136 through 151 )
7X-RAY DIFFRACTION7chain 'A' and (resid 152 through 163 )
8X-RAY DIFFRACTION8chain 'A' and (resid 164 through 188 )
9X-RAY DIFFRACTION9chain 'A' and (resid 189 through 205 )
10X-RAY DIFFRACTION10chain 'A' and (resid 206 through 220 )
11X-RAY DIFFRACTION11chain 'B' and (resid 12 through 37 )
12X-RAY DIFFRACTION12chain 'B' and (resid 38 through 70 )
13X-RAY DIFFRACTION13chain 'B' and (resid 71 through 94 )
14X-RAY DIFFRACTION14chain 'B' and (resid 95 through 103 )
15X-RAY DIFFRACTION15chain 'B' and (resid 104 through 136 )
16X-RAY DIFFRACTION16chain 'B' and (resid 137 through 160 )
17X-RAY DIFFRACTION17chain 'B' and (resid 161 through 178 )

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