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- PDB-3cd9: Development of a family of redox-sensitive green fluorescent prot... -

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Basic information

Entry
Database: PDB / ID: 3cd9
TitleDevelopment of a family of redox-sensitive green fluorescent protein indicators for use in relatively oxidizing subcellular environments
ComponentsGreen fluorescent protein
KeywordsLUMINESCENT PROTEIN / GFP / redox
Function / homology
Function and homology information


bioluminescence / generation of precursor metabolites and energy
Similarity search - Function
Green Fluorescent Protein / Green fluorescent protein / Green fluorescent protein, GFP / Green fluorescent protein-related / Green fluorescent protein / Green fluorescent protein / Beta Barrel / Mainly Beta
Similarity search - Domain/homology
Green fluorescent protein
Similarity search - Component
Biological speciesAequorea victoria (jellyfish)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 1.502 Å
AuthorsLohman, J.R.
CitationJournal: Biochemistry / Year: 2008
Title: Development of a family of redox-sensitive green fluorescent protein indicators for use in relatively oxidizing subcellular environments
Authors: Lohman, J.R. / Remington, S.J.
History
DepositionFeb 26, 2008Deposition site: RCSB / Processing site: RCSB
Revision 1.0Aug 19, 2008Provider: repository / Type: Initial release
Revision 1.1Jul 13, 2011Group: Version format compliance
Revision 1.2Oct 20, 2021Group: Database references / Derived calculations / Category: database_2 / struct_conn / struct_ref_seq_dif
Item: _database_2.pdbx_DOI / _database_2.pdbx_database_accession ..._database_2.pdbx_DOI / _database_2.pdbx_database_accession / _struct_conn.pdbx_leaving_atom_flag / _struct_ref_seq_dif.details
Revision 1.3Nov 6, 2024Group: Data collection / Structure summary
Category: chem_comp_atom / chem_comp_bond ...chem_comp_atom / chem_comp_bond / pdbx_entry_details / pdbx_modification_feature

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

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Assembly

Deposited unit
A: Green fluorescent protein


Theoretical massNumber of molelcules
Total (without water)28,1391
Polymers28,1391
Non-polymers00
Water4,396244
1


  • Idetical with deposited unit
  • defined by author&software
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
MethodPISA
Unit cell
Length a, b, c (Å)51.174, 62.353, 69.618
Angle α, β, γ (deg.)90.00, 90.00, 90.00
Int Tables number19
Space group name H-MP212121

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Components

#1: Protein Green fluorescent protein / roGFP1-iE / GFP


Mass: 28138.557 Da / Num. of mol.: 1
Mutation: C48S,F64L,F99S,S147CR,H148S,M153T,V163A,I167T,Q204C
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Aequorea victoria (jellyfish) / Gene: GFP / Plasmid: pQE-30 / Production host: Escherichia coli (E. coli) / Strain (production host): DH5 alpha / References: UniProt: P42212
#2: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 244 / Source method: isolated from a natural source / Formula: H2O
Has protein modificationY

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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: 1.97 Å3/Da / Density % sol: 37.68 %
Crystal growTemperature: 298 K / Method: vapor diffusion, hanging drop / pH: 8
Details: 0.1M tris, 24% PEG 1550, 0.04M sodium acetate. TCEP added to 20mM after crystal formation., pH 8.0, VAPOR DIFFUSION, HANGING DROP, temperature 298K

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: ALS / Beamline: 5.0.1 / Wavelength: 1 Å
DetectorType: ADSC QUANTUM 210 / Detector: CCD / Date: Aug 11, 2007
RadiationMonochromator: Single crystal, cylindrically bent, Si(220) / Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 1 Å / Relative weight: 1
ReflectionResolution: 1.502→50 Å / Num. all: 35231 / Num. obs: 35231 / % possible obs: 97.2 % / Observed criterion σ(I): 2
Reflection shellResolution: 1.502→1.55 Å / % possible all: 95.1

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Processing

Software
NameVersionClassification
ADSCQuantumdata collection
MOLREPphasing
REFMAC5.2refinement
HKL-2000data reduction
HKL-2000data scaling
RefinementMethod to determine structure: MOLECULAR REPLACEMENT / Resolution: 1.502→30 Å / Cor.coef. Fo:Fc: 0.968 / Cor.coef. Fo:Fc free: 0.96 / SU B: 2.723 / SU ML: 0.047 / Cross valid method: THROUGHOUT / ESU R: 0.096 / ESU R Free: 0.074 / Stereochemistry target values: MAXIMUM LIKELIHOOD / Details: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS
RfactorNum. reflection% reflectionSelection details
Rfree0.19308 1769 5 %RANDOM
Rwork0.16715 ---
all0.16847 33419 --
obs0.16847 33419 97.17 %-
Solvent computationIon probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.2 Å / Solvent model: MASK
Displacement parametersBiso mean: 14.445 Å2
Baniso -1Baniso -2Baniso -3
1--0.1 Å20 Å20 Å2
2---0.69 Å20 Å2
3---0.79 Å2
Refinement stepCycle: LAST / Resolution: 1.502→30 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms1809 0 0 244 2053
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.010.0221855
X-RAY DIFFRACTIONr_bond_other_d0.0010.021246
X-RAY DIFFRACTIONr_angle_refined_deg1.4691.9692515
X-RAY DIFFRACTIONr_angle_other_deg0.82533052
X-RAY DIFFRACTIONr_dihedral_angle_1_deg6.2625233
X-RAY DIFFRACTIONr_dihedral_angle_2_deg34.48225.11488
X-RAY DIFFRACTIONr_dihedral_angle_3_deg11.7215312
X-RAY DIFFRACTIONr_dihedral_angle_4_deg7.21157
X-RAY DIFFRACTIONr_chiral_restr0.080.2274
X-RAY DIFFRACTIONr_gen_planes_refined0.0050.022086
X-RAY DIFFRACTIONr_gen_planes_other0.0010.02368
X-RAY DIFFRACTIONr_nbd_refined0.2020.2263
X-RAY DIFFRACTIONr_nbd_other0.1980.21194
X-RAY DIFFRACTIONr_nbtor_refined0.1670.2858
X-RAY DIFFRACTIONr_nbtor_other0.1110.2983
X-RAY DIFFRACTIONr_xyhbond_nbd_refined0.1480.2141
X-RAY DIFFRACTIONr_xyhbond_nbd_other
X-RAY DIFFRACTIONr_metal_ion_refined
X-RAY DIFFRACTIONr_metal_ion_other
X-RAY DIFFRACTIONr_symmetry_vdw_refined0.0660.22
X-RAY DIFFRACTIONr_symmetry_vdw_other0.3210.222
X-RAY DIFFRACTIONr_symmetry_hbond_refined0.0840.217
X-RAY DIFFRACTIONr_symmetry_hbond_other
X-RAY DIFFRACTIONr_symmetry_metal_ion_refined
X-RAY DIFFRACTIONr_symmetry_metal_ion_other
X-RAY DIFFRACTIONr_mcbond_it0.9991.51182
X-RAY DIFFRACTIONr_mcbond_other0.3191.5468
X-RAY DIFFRACTIONr_mcangle_it1.47121834
X-RAY DIFFRACTIONr_scbond_it2.0733782
X-RAY DIFFRACTIONr_scangle_it2.9084.5676
X-RAY DIFFRACTIONr_rigid_bond_restr1.1933454
X-RAY DIFFRACTIONr_sphericity_free4.3673244
X-RAY DIFFRACTIONr_sphericity_bonded1.8633055
LS refinement shellResolution: 1.502→1.541 Å / Total num. of bins used: 20
RfactorNum. reflection% reflection
Rfree0.29 125 -
Rwork0.205 2336 -
obs--93.65 %

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