4GL7
Structure of human placental aromatase complexed with designed inhibitor HDDG046 (compound 5)
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | SYNCHROTRON |
Source details | CHESS BEAMLINE A1 |
Synchrotron site | CHESS |
Beamline | A1 |
Temperature [K] | 100 |
Detector technology | CCD |
Collection date | 2011-11-12 |
Detector | ADSC QUANTUM 210 |
Wavelength(s) | 0.978 |
Spacegroup name | P 32 2 1 |
Unit cell lengths | 141.445, 141.445, 118.851 |
Unit cell angles | 90.00, 90.00, 120.00 |
Refinement procedure
Resolution | 50.000 - 3.900 |
R-factor | 0.21615 |
Rwork | 0.214 |
R-free | 0.25407 |
Structure solution method | FOURIER SYNTHESIS |
RMSD bond length | 0.012 |
RMSD bond angle | 1.474 |
Data reduction software | HKL-2000 |
Data scaling software | HKL-2000 |
Phasing software | CCP4 |
Refinement software | REFMAC (5.5.0109) |
Data quality characteristics
Overall | |
Low resolution limit [Å] | 122.490 |
High resolution limit [Å] | 3.900 |
Rmerge | 0.114 |
Number of reflections | 12126 |
<I/σ(I)> | 16.7 |
Completeness [%] | 99.5 |
Redundancy | 4.6 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, SITTING DROP | 7.4 | 277 | The enzyme-inhibitor complexes were prepared by the addition from 20mM stock solutions of compound 5 in PEG550 to 18 M (~1mg/ml) of aromatase, to give a final inhibitor concentration of 300 microM. The mixture was incubated overnight at 4 C in 100mM potassium phosphate buffer pH 7.4 containing 20% glycerol, 20mM dithiothreitol, 0.5 microM 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, SITTING DROP, temperature 277K |