X-RAY DIFFRACTION

Starting Model(s)

Initial Refinement Model(s)
TypeSourceAccession CodeDetails
experimental modelPDB 1MD9Mutated DhbE (pdb code 1MD9) with C-terminal domain rotated and pdb entry code 2FQ1
experimental modelPDB 2FQ1Mutated DhbE (pdb code 1MD9) with C-terminal domain rotated and pdb entry code 2FQ1

Crystallization

Crystalization Experiments
IDMethodpHTemperatureDetails
1microbatch under oil62931:1 mixture of protein and 20-30% PEG 3350, 100 mM NH4I, 50 mM MES , pH 6.0, microbatch under oil, temperature 293K
Crystal Properties
Matthews coefficientSolvent content
2.7555.3

Crystal Data

Unit Cell
Length ( Å )Angle ( ˚ )
a = 160.575α = 90
b = 101.771β = 107.06
c = 240.678γ = 90
Symmetry
Space GroupP 1 21 1

Diffraction

Diffraction Experiment
ID #Crystal IDScattering TypeData Collection TemperatureDetectorDetector TypeDetailsCollection DateMonochromatorProtocol
11x-ray113CCDMARMOSAIC 325 mm CCDSSRL 11-12007-12-11MSINGLE WAVELENGTH
Radiation Source
ID #SourceTypeWavelength ListSynchrotron SiteBeamline
1SYNCHROTRONSSRL BEAMLINE BL11-10.97945SSRLBL11-1

Data Collection

Overall
ID #Resolution (High)Resolution (Low)Percent Possible (Observed)R Merge I (Observed)Net I Over Average Sigma (I)RedundancyNumber Reflections (All)Number Reflections (Observed)Observed Criterion Sigma (F)Observed Criterion Sigma (I)B (Isotropic) From Wilson Plot
13.14099.40.068133.7135141134330-390
Highest Resolution Shell
ID #Resolution (High)Resolution (Low)Percent Possible (All)Percent Possible (Observed)R Merge I (Observed)Mean I Over Sigma (Observed)RedundancyNumber Unique Reflections (All)
13.13.21000.4741.83.712070

Refinement

Statistics
Diffraction IDStructure Solution MethodStarting modelResolution (High)Resolution (Low)Cut-off Sigma (F)Number Reflections (All)Number Reflections (Observed)Number Reflections (R-Free)Percent Reflections (Observed)R-Factor (Observed)R-WorkR-FreeR-Free Selection DetailsMean Isotropic B
X-RAY DIFFRACTIONMOLECULAR REPLACEMENTMutated DhbE (pdb code 1MD9) with C-terminal domain rotated and pdb entry code 2FQ13.139.2691.34135717134306638598.960.22030.21790.2639Rfree chosen in thin shells with SHELX
Temperature Factor Modeling
Anisotropic B[1][1]Anisotropic B[1][2]Anisotropic B[1][3]Anisotropic B[2][2]Anisotropic B[2][3]Anisotropic B[3][3]
1.4217-0.60873.9582-5.3798
RMS Deviations
KeyRefinement Restraint Deviation
f_dihedral_angle_d15.753
f_angle_d1.356
f_chiral_restr0.084
f_bond_d0.01
f_plane_restr0.006
Non-Hydrogen Atoms Used in Refinement
Non-Hydrogen AtomsNumber
Protein Atoms44567
Nucleic Acid Atoms
Solvent Atoms
Heterogen Atoms545

Software

Software
Software NamePurpose
PHENIXrefinement
PHASERphasing
BALBESphasing
REFMACrefinement
HKL-2000data collection
HKL-2000data reduction
HKL-2000data scaling