3L08

Structure of Pi3K gamma with a potent inhibitor: GSK2126458

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 21-ID-F
Synchrotron site	APS
Beamline	21-ID-F
Detector technology	CCD
Collection date	2008-07-01
Detector	MARMOSAIC 225 mm CCD
Wavelength(s)	0.97872
Spacegroup name	C 1 2 1
Unit cell lengths	145.617, 68.410, 107.470
Unit cell angles	90.00, 94.72, 90.00

Refinement procedure

Resolution	41.215 - 2.700
R-factor	0.2008
Rwork	0.198
R-free	0.26140
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	Proprietary pi3K gamma structure
RMSD bond length	0.004
RMSD bond angle	0.731
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	PHENIX
Refinement software	PHENIX ((phenix.refine: 1.4_87))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	50.000	2.800
High resolution limit [Å]	2.700	2.700
Number of reflections	28636
<I/σ(I)>	10.4	1.9
Completeness [%]	98.0	83.1

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP		277	Protein was at 7.9mg/ml in 20mM Tris pH 7.2, 50mM ammonium sulfate, 1% ethylene glycol, 1% betaine, 0.02% CHAPS, 5mM DTT with 1mM inhbitor reacted overnight at 4C. 2 microliter protein solution plus 2 microliters well in sitting drops. Well consisted of 20% PEG 3350, 0.1M Tris pH 8.0, 3% 1,6 hexanediol, 0.2M ammonium sulfate. Crystals grew from dilution seeding from apo seeds. Cryo was 25% ethylene glycol mixed with the well, VAPOR DIFFUSION, SITTING DROP, temperature 277K