3QI3
Crystal structure of PDE9A(Q453E) in complex with inhibitor BAY73-6691
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | SYNCHROTRON |
Source details | NSLS BEAMLINE X29A |
Synchrotron site | NSLS |
Beamline | X29A |
Temperature [K] | 100 |
Detector technology | CCD |
Detector | ADSC QUANTUM 315 |
Wavelength(s) | 1.1 |
Spacegroup name | P 41 21 2 |
Unit cell lengths | 103.127, 103.127, 270.441 |
Unit cell angles | 90.00, 90.00, 90.00 |
Refinement procedure
Resolution | 30.000 - 2.300 |
R-factor | 0.222 |
Rwork | 0.222 |
R-free | 0.24200 |
Structure solution method | MOLECULAR REPLACEMENT |
RMSD bond length | 0.007 |
RMSD bond angle | 1.200 |
Data reduction software | HKL-2000 |
Data scaling software | SCALEPACK |
Phasing software | AMoRE |
Refinement software | CNS |
Data quality characteristics
Overall | |
Low resolution limit [Å] | 30.000 |
High resolution limit [Å] | 2.300 |
Rmerge | 0.092 |
Number of reflections | 63252 |
Completeness [%] | 95.7 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, HANGING DROP | 7.5 | 277 | The catalytic domain of the PDE9Q453E mutant (10-15 mg/mL, amino acids 181-506) was stored in a buffer of 50 mM NaCl, 20 mM Tris.HCl pH 7.5, 1 mM beta-mercaptoethanol, and 1 mM EDTA. After mixing with 2 mM IBMX, the PDE9Q453E-IBMX complex was crystallized by hanging drop vapor diffusion against the well buffer of 2.0 M Na formate, 0.1 M HEPES pH 7.5, 5% xylitol at 277K. Crystals of the PDE9Q453E-(S)-BAY73-6691 complex were prepared by soaking PDE9Q453E-IBMX co-crystals in the crystallization buffer plus 2 mM (S)-BAY73-6691 at 398K for 3 days, VAPOR DIFFUSION, HANGING DROP |