4YZC
Crystal structure of pIRE1alpha in complex with staurosporine
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | SYNCHROTRON |
Source details | APS BEAMLINE 21-ID-D |
Synchrotron site | APS |
Beamline | 21-ID-D |
Temperature [K] | 100 |
Detector technology | CCD |
Collection date | 2012-02-17 |
Detector | ADSC QUANTUM 315 |
Wavelength(s) | 0.97872 |
Spacegroup name | P 21 21 21 |
Unit cell lengths | 49.337, 155.260, 155.643 |
Unit cell angles | 90.00, 90.00, 90.00 |
Refinement procedure
Resolution | 40.225 - 2.494 |
R-factor | 0.2489 |
Rwork | 0.247 |
R-free | 0.28920 |
Structure solution method | MOLECULAR REPLACEMENT |
Starting model (for MR) | 3p23 |
RMSD bond length | 0.003 |
RMSD bond angle | 0.648 |
Data scaling software | SCALEPACK |
Phasing software | MOLREP |
Refinement software | PHENIX (1.9_1692) |
Data quality characteristics
Overall | Outer shell | |
Low resolution limit [Å] | 40.230 | 2.583 |
High resolution limit [Å] | 2.494 | 2.494 |
Rmerge | 0.097 | 0.134 |
Total number of observations | 226940 | |
Number of reflections | 41927 | |
<I/σ(I)> | 13.36 | |
Completeness [%] | 98.2 | 88.06 |
Redundancy | 6.5 | 6.2 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, SITTING DROP | 7.5 | 293 | The complex of pIRE1a (547-977) with staurosporine was prepared by mixing the protein with 0.5 mM staurosporine and incubated overnight on ice. The crystals were grown at 20C by vapor diffusion in sitting drop containing 2 ul of protein (13 mg/ml in 50 mM Hepes, pH 7.5, 200 mM NaCl, 5 mM DTT, 1 mM EDTA) and 2ul reservoir solution containing PEG 300 (30%-40%), 100 mM Hepes pH 7.5, 200 mM KCl. Small hexagonal plates appeared over 5-10 days and reached full size (0.05 X 0.75 X 0.15 mm) in ~20 days. The crystals were flash-frozen in liquid N2 directly from the crystallization drop. All diffraction data was collected at the Advanced Photon Source, Argonne National Laboratories, Life Sciences CAT, Sector 21 |