1OJJ

Anatomy of glycosynthesis: Structure and kinetics of the Humicola insolens Cel7BE197A and E197S glycosynthase mutants


X-RAY DIFFRACTION

Starting Model(s)

Initial Refinement Model(s)
TypeSourceAccession CodeDetails
experimental modelPDB 1DYMPDB ENTRY 1DYM

Crystallization

Crystalization Experiments
IDMethodpHTemperatureDetails
1VAPOR DIFFUSION, HANGING DROP7HANGING DROPS 20MM TRIS-HCL PH7-8.5, 280 15-30% POLYETHYLENE GLYCOL 4000, pH 7.00
Crystal Properties
Matthews coefficientSolvent content
2.447.5

Crystal Data

Unit Cell
Length ( Å )Angle ( ˚ )
a = 66.364α = 90
b = 74.746β = 102.51
c = 85.82γ = 90
Symmetry
Space GroupP 1 21 1

Diffraction

Diffraction Experiment
ID #Crystal IDScattering TypeData Collection TemperatureDetectorDetector TypeDetailsCollection DateMonochromatorProtocol
11x-ray100CCDMARRESEARCHMIRRORSMSINGLE WAVELENGTH
Radiation Source
ID #SourceTypeWavelength ListSynchrotron SiteBeamline
1SYNCHROTRONESRF BEAMLINE ID14-1ESRFID14-1

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
11.415990.033353.6157771
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)
11.41.45960.33.53.2

Refinement

Statistics
Diffraction IDStructure Solution MethodCross Validation methodStarting modelResolution (High)Resolution (Low)Number Reflections (Observed)Number Reflections (R-Free)Percent Reflections (Observed)R-Factor (Observed)R-WorkR-FreeR-Free Selection DetailsMean Isotropic B
X-RAY DIFFRACTIONMOLECULAR REPLACEMENTTHROUGHOUTPDB ENTRY 1DYM1.484.52149738794598.70.150.1490.173RANDOM11.65
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]
0.17-0.25-0.310.03
RMS Deviations
KeyRefinement Restraint Deviation
r_dihedral_angle_2_deg31.314
r_dihedral_angle_4_deg17.231
r_dihedral_angle_3_deg12.129
r_dihedral_angle_1_deg6.234
r_scangle_it3.398
r_scbond_it2.602
r_mcangle_it1.664
r_angle_refined_deg1.569
r_mcbond_it1.426
r_angle_other_deg0.848
RMS Deviations
KeyRefinement Restraint Deviation
r_dihedral_angle_2_deg31.314
r_dihedral_angle_4_deg17.231
r_dihedral_angle_3_deg12.129
r_dihedral_angle_1_deg6.234
r_scangle_it3.398
r_scbond_it2.602
r_mcangle_it1.664
r_angle_refined_deg1.569
r_mcbond_it1.426
r_angle_other_deg0.848
r_mcbond_other0.472
r_nbd_refined0.213
r_symmetry_vdw_refined0.211
r_symmetry_hbond_refined0.208
r_nbd_other0.189
r_symmetry_vdw_other0.179
r_chiral_restr0.096
r_xyhbond_nbd_refined0.089
r_nbtor_other0.083
r_bond_refined_d0.012
r_gen_planes_refined0.007
r_bond_other_d0.001
r_gen_planes_other0.001
r_nbtor_refined
r_xyhbond_nbd_other
r_metal_ion_refined
r_metal_ion_other
r_symmetry_hbond_other
r_symmetry_metal_ion_refined
r_symmetry_metal_ion_other
r_mcangle_other
r_scbond_other
r_scangle_other
r_long_range_B_refined
r_long_range_B_other
r_rigid_bond_restr
r_sphericity_free
r_sphericity_bonded
Non-Hydrogen Atoms Used in Refinement
Non-Hydrogen AtomsNumber
Protein Atoms6170
Nucleic Acid Atoms
Solvent Atoms642
Heterogen Atoms120

Software

Software
Software NamePurpose
REFMACrefinement
DENZOdata reduction
SCALEPACKdata scaling
AMoREphasing