2IB8

Crystallographic and kinetic studies of human mitochondrial acetoacetyl-CoA thiolase (T2): the importance of potassium and chloride for its structure and function


X-RAY DIFFRACTION

Starting Model(s)

Initial Refinement Model(s)
TypeSourceAccession CodeDetails
experimental modelPDB 1WL4PDB entry 1WL4

Crystallization

Crystalization Experiments
IDMethodpHTemperatureDetails
1VAPOR DIFFUSION, HANGING DROP6.527718% PEG 5000 monomethylether, 0.1M MES, pH 6.5, VAPOR DIFFUSION, HANGING DROP, temperature 277.0K
Crystal Properties
Matthews coefficientSolvent content
2.4349.51

Crystal Data

Unit Cell
Length ( Å )Angle ( ˚ )
a = 75.651α = 90
b = 107.277β = 103.07
c = 102.209γ = 90
Symmetry
Space GroupP 1 21 1

Diffraction

Diffraction Experiment
ID #Crystal IDScattering TypeData Collection TemperatureDetectorDetector TypeDetailsCollection DateMonochromatorProtocol
11x-ray100CCDMAR CCD 165 mmmirrors2005-12-17MSINGLE WAVELENGTH
Radiation Source
ID #SourceTypeWavelength ListSynchrotron SiteBeamline
1SYNCHROTRONMAX II BEAMLINE I7111.141MAX III711

Data Collection

Overall
ID #Resolution (High)Resolution (Low)Percent Possible (Observed)R Merge I (Observed)R Sym 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
11.8547.2599.90.0610.06116.94.213535013525724
Highest Resolution Shell
ID #Resolution (High)Resolution (Low)Percent Possible (All)Percent Possible (Observed)R Merge I (Observed)R-Sym I (Observed)Mean I Over Sigma (Observed)RedundancyNumber Unique Reflections (All)
11.851.951000.3010.3014.64.119653

Refinement

Statistics
Diffraction IDStructure Solution MethodCross Validation methodStarting modelResolution (High)Resolution (Low)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 REPLACEMENTTHROUGHOUTPDB entry 1WL41.8547.2513525912849667631000.164320.162170.20542RANDOM24.195
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]
-0.21-0.420.3-0.28
RMS Deviations
KeyRefinement Restraint Deviation
r_dihedral_angle_2_deg40.719
r_dihedral_angle_4_deg13.463
r_dihedral_angle_3_deg13.235
r_dihedral_angle_1_deg5.664
r_scangle_it2.281
r_mcangle_it2.043
r_scbond_it1.65
r_mcbond_it1.404
r_angle_refined_deg1.304
r_nbtor_refined0.294
RMS Deviations
KeyRefinement Restraint Deviation
r_dihedral_angle_2_deg40.719
r_dihedral_angle_4_deg13.463
r_dihedral_angle_3_deg13.235
r_dihedral_angle_1_deg5.664
r_scangle_it2.281
r_mcangle_it2.043
r_scbond_it1.65
r_mcbond_it1.404
r_angle_refined_deg1.304
r_nbtor_refined0.294
r_nbd_refined0.205
r_symmetry_vdw_refined0.186
r_xyhbond_nbd_refined0.127
r_symmetry_hbond_refined0.112
r_chiral_restr0.092
r_metal_ion_refined0.069
r_bond_refined_d0.013
r_gen_planes_refined0.005
Non-Hydrogen Atoms Used in Refinement
Non-Hydrogen AtomsNumber
Protein Atoms11512
Nucleic Acid Atoms
Solvent Atoms1033
Heterogen Atoms86

Software

Software
Software NamePurpose
REFMACrefinement
HKL-2000data collection
XDSdata reduction
XDSdata scaling
MOLREPphasing