Crystal structure of vitamin D hydroxylase cytochrome P450 105A1 (wild type) with imidazole bound
X-RAY DIFFRACTION
Crystallization
Crystalization Experiments | ||||
---|---|---|---|---|
ID | Method | pH | Temperature | Details |
1 | VAPOR DIFFUSION, SITTING DROP | 6.2 | 283 | 24% PEG 8000, 0.1M imidazole, 0.2M sodium chloride, 0.2M sodium formate, pH 6.2, VAPOR DIFFUSION, SITTING DROP, temperature 283K |
Crystal Properties | |
---|---|
Matthews coefficient | Solvent content |
2.21 | 44.32 |
Crystal Data
Unit Cell | |
---|---|
Length ( Å ) | Angle ( ˚ ) |
a = 52.575 | α = 90 |
b = 53.754 | β = 90 |
c = 140.948 | γ = 90 |
Symmetry | |
---|---|
Space Group | P 21 21 21 |
Diffraction
Diffraction Experiment | ||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
ID # | Crystal ID | Scattering Type | Data Collection Temperature | Detector | Detector Type | Details | Collection Date | Monochromator | Protocol | |||||
1 | 1 | x-ray | 90 | CCD | ADSC QUANTUM 210 | mirrors | 2006-09-23 | M | SINGLE WAVELENGTH | |||||
2 | 1 | x-ray | 90 | CCD | ADSC QUANTUM 210 | mirrors | 2005-10-20 | M | MAD |
Radiation Source | |||||
---|---|---|---|---|---|
ID # | Source | Type | Wavelength List | Synchrotron Site | Beamline |
1 | SYNCHROTRON | SPRING-8 BEAMLINE BL44B2 | 1.0000 | SPring-8 | BL44B2 |
2 | SYNCHROTRON | SPRING-8 BEAMLINE BL44B2 | 1.7377, 1.7403, 1.7310, 1.7240 | SPring-8 | BL44B2 |
Data Collection
Overall | |||||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
ID # | Resolution (High) | Resolution (Low) | Percent Possible (Observed) | R Sym I (Observed) | Net I Over Average Sigma (I) | Redundancy | Number Reflections (All) | Number Reflections (Observed) | Observed Criterion Sigma (F) | Observed Criterion Sigma (I) | B (Isotropic) From Wilson Plot | ||||||||
1,2 | 1.5 | 20 | 97.1 | 0.035 | 32.3 | 9 | 62925 | -3 | 25.7 |
Highest Resolution Shell | |||||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
ID # | Resolution (High) | Resolution (Low) | Percent Possible (All) | Percent Possible (Observed) | R-Sym I (Observed) | Mean I Over Sigma (Observed) | Redundancy | Number Unique Reflections (All) | |||||||||||
1.5 | 1.53 | 91.1 | 0.264 | 7.4 | 8.1 | 2863 |
Refinement
Statistics | |||||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Diffraction ID | Structure Solution Method | Cross Validation method | Starting model | Resolution (High) | Resolution (Low) | Number Reflections (All) | Number Reflections (Observed) | Number Reflections (R-Free) | Percent Reflections (Observed) | R-Factor (All) | R-Factor (Observed) | R-Work (Depositor) | R-Work (DCC) | R-Free (Depositor) | R-Free (DCC) | R-Free Selection Details | Mean Isotropic B | ||
X-RAY DIFFRACTION | MAD and MOLECULAR REPLACEMENT | THROUGHOUT | Starting model was obtained by MAD phasing using iron atom as a anomalous scatterer | 1.5 | 19.62 | 59532 | 59532 | 3180 | 96.82 | 0.19578 | 0.19578 | 0.19434 | 0.19 | 0.22327 | 0.22 | RANDOM | 18.555 |
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] | |
0.7 | -0.61 | -0.09 |
RMS Deviations | |
---|---|
Key | Refinement Restraint Deviation |
r_dihedral_angle_2_deg | 34.441 |
r_dihedral_angle_4_deg | 15.114 |
r_dihedral_angle_3_deg | 12.269 |
r_dihedral_angle_1_deg | 4.865 |
r_scangle_it | 2.788 |
r_scbond_it | 1.852 |
r_angle_refined_deg | 1.254 |
r_mcangle_it | 1.167 |
r_mcbond_it | 0.739 |
r_nbtor_refined | 0.304 |
Non-Hydrogen Atoms Used in Refinement | |
---|---|
Non-Hydrogen Atoms | Number |
Protein Atoms | 3072 |
Nucleic Acid Atoms | |
Solvent Atoms | 524 |
Heterogen Atoms | 48 |
Software
Software | |
---|---|
Software Name | Purpose |
REFMAC | refinement |
BSS | data collection |
HKL-2000 | data reduction |
HKL-2000 | data scaling |
SHARP | phasing |