4ANX
Complexes of PI3Kgamma with isoform selective inhibitors.
X-RAY DIFFRACTION
Crystallization
| Crystalization Experiments | ||||
|---|---|---|---|---|
| ID | Method | pH | Temperature | Details |
| 1 | VAPOR DIFFUSION, HANGING DROP | 7.5 | 292 | CRYSTALS WERE GROWN AT 19C IN HANGING DROPS USING VAPOR DIFFUSION. THE COMPLEX WAS FORMED BY INCUBATING 49.5UL OF PI3KG-HIS6 (7.2 MG/ML IN 20 MM TRIS-HCL, PH 7.2, 50 MM (NH4)2SO4, AND 1 MM TCEP) WITH 0.5UL OF LIGAND (100 MM STOCK IN 100% DMSO) FOR 30 MINUTES AT 4C. CRYSTALS OF THE BINARY COMPLEX WERE OBTAINED BY MIXING 1.0UL OF THE PI3KG:INHIBITOR COMPLEX WITH 1.0UL OF A RESERVOIR SOLUTION CONSISTING OF 16-21% PEG 4K, 0.25 M (NH4)2SO4, AND 0.1 M TRIS-HCL, PH 7.5. |
| Crystal Properties | |
|---|---|
| Matthews coefficient | Solvent content |
| 2.32 | 47 |
Crystal Data
| Unit Cell | |
|---|---|
| Length ( Å ) | Angle ( ˚ ) |
| a = 143.424 | α = 90 |
| b = 68.531 | β = 95.26 |
| c = 106.263 | γ = 90 |
| Symmetry | |
|---|---|
| Space Group | C 1 2 1 |
Diffraction
| Diffraction Experiment | ||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| ID # | Crystal ID | Scattering Type | Data Collection Temperature | Detector | Detector Type | Details | Collection Date | Monochromator | Protocol | |||||
| 1 | 1 | x-ray | 155 | CCD | ADSC QUANTUM 315r | 2010-02-24 | M | SINGLE WAVELENGTH | ||||||
| Radiation Source | |||||
|---|---|---|---|---|---|
| ID # | Source | Type | Wavelength List | Synchrotron Site | Beamline |
| 1 | SYNCHROTRON | SSRL BEAMLINE BL7-1 | SSRL | BL7-1 | |
Data Collection
| Overall | |||||||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| ID # | Resolution (High) | Resolution (Low) | Percent Possible (Observed) | R Merge 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 | 2.73 | 32.6 | 99.4 | 0.08 | 14.3 | 3.7 | 27406 | -3 | |||||||||||
| Highest Resolution Shell | |||||||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| ID # | Resolution (High) | Resolution (Low) | Percent Possible (All) | Percent Possible (Observed) | R Merge I (Observed) | Mean I Over Sigma (Observed) | Redundancy | Number Unique Reflections (All) | |||||||||||
| 1 | 2.73 | 3.07 | 99.4 | 0.77 | 1.8 | 3.7 | |||||||||||||
Refinement
| Statistics | |||||||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Diffraction ID | Structure Solution Method | Cross Validation method | Starting model | Resolution (High) | Resolution (Low) | Number Reflections (Observed) | Number Reflections (R-Free) | Percent Reflections (Observed) | R-Factor (Observed) | R-Work | R-Free | R-Free Selection Details | Mean Isotropic B | ||||||
| X-RAY DIFFRACTION | MOLECULAR REPLACEMENT | THROUGHOUT | IN-HOUSE PI3KG STRUCTURE | 2.73 | 32.6 | 26019 | 1375 | 99.45 | 0.22771 | 0.22472 | 0.28187 | RANDOM | |||||||
| 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] | |
| 0.01 | 0.07 | 0.04 | -0.03 | |||
| RMS Deviations | |
|---|---|
| Key | Refinement Restraint Deviation |
| r_dihedral_angle_2_deg | 37.075 |
| r_dihedral_angle_3_deg | 19.999 |
| r_dihedral_angle_4_deg | 15.206 |
| r_dihedral_angle_1_deg | 7.916 |
| r_scangle_it | 2.409 |
| r_scbond_it | 1.682 |
| r_angle_refined_deg | 1.601 |
| r_mcangle_it | 1.277 |
| r_angle_other_deg | 0.996 |
| r_mcbond_it | 0.943 |
| Non-Hydrogen Atoms Used in Refinement | |
|---|---|
| Non-Hydrogen Atoms | Number |
| Protein Atoms | 6757 |
| Nucleic Acid Atoms | |
| Solvent Atoms | 34 |
| Heterogen Atoms | 37 |
Software
| Software | |
|---|---|
| Software Name | Purpose |
| REFMAC | refinement |
| XDS | data reduction |
| XSCALE | data scaling |
| PHASER | phasing |
