4GL5
Structure of human placental aromatase complexed with designed inhibitor HDDG029 (compound 4)
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | SYNCHROTRON |
Source details | APS BEAMLINE 19-ID |
Synchrotron site | APS |
Beamline | 19-ID |
Temperature [K] | 100 |
Detector technology | CCD |
Collection date | 2009-12-18 |
Detector | ADSC QUANTUM 315 |
Wavelength(s) | 0.979 |
Spacegroup name | P 32 2 1 |
Unit cell lengths | 140.329, 140.329, 118.733 |
Unit cell angles | 90.00, 90.00, 120.00 |
Refinement procedure
Resolution | 50.000 - 3.480 |
R-factor | 0.21277 |
Rwork | 0.210 |
R-free | 0.26036 |
Structure solution method | FOURIER SYNTHESIS |
RMSD bond length | 0.009 |
RMSD bond angle | 1.311 |
Data reduction software | HKL-3000 |
Data scaling software | HKL-3000 |
Phasing software | CCP4 |
Refinement software | REFMAC (5.5.0109) |
Data quality characteristics
Overall | |
Low resolution limit [Å] | 121.530 |
High resolution limit [Å] | 3.480 |
Rmerge | 0.154 |
Number of reflections | 17289 |
<I/σ(I)> | 14.1 |
Completeness [%] | 91.5 |
Redundancy | 3.8 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION | 7.4 | 277 | The enzyme-inhibitor complexes were prepared by the addition from 20mM stock solutions of compound 4 in PEG550 to 18 M (~1mg/ml) of aromatase, to give a final inhibitor concentration of 300 uM. The mixture was incubated overnight at 4 C in 100mM potassium phosphate buffer pH 7.4 containing 20% glycerol, 20mM dithiothreitol, 0.5 M ASD and 1mM BDM. The complex was then concentrated to 25-30mg/ml using ultrafiltration. Protein was setup for crystallization using protein to cocktail ratios 2:1 and 3:1. The protein was mixed with reservoir cocktails of 24-30% polyethylene glycol 4000 in 50mM NaCl, 50mM Tris, pH 8.5 and vapor diffused in sealed 24-well sitting drop plates against corresponding reservoir solution., VAPOR DIFFUSION, temperature 277K |