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