4ANW

Complexes of PI3Kgamma with isoform selective inhibitors.

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 21-ID-F
Synchrotron site	APS
Beamline	21-ID-F
Temperature [K]	155
Detector technology	CCD
Collection date	2010-08-24
Detector	ADSC CCD
Spacegroup name	C 1 2 1
Unit cell lengths	143.424, 68.531, 106.263
Unit cell angles	90.00, 95.26, 90.00

Refinement procedure

Resolution	71.430 - 2.310
R-factor	0.238
Rwork	0.235
R-free	0.29600
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	IN-HOUSE PI3KG STRUCTURE
RMSD bond length	0.019
RMSD bond angle	1.831
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	PHASER
Refinement software	REFMAC (5.2.0019)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	71.400	2.590
High resolution limit [Å]	2.310	2.310
Rmerge	0.060	0.850
Number of reflections	44953
<I/σ(I)>	15.1	1.86
Completeness [%]	99.2	99.4
Redundancy	3.7	3.8

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7.5	292	CRYSTALS WERE GROWN AT 19C IN HANGING DROPS USING VAPOR DIFFUSION. THE COMPLEX WAS FORMED BY INCUBATING 49.5UL OF PI3KG-HIS6 (7.2 MG/ML IN 20 MM TRIS-HCL, PH 7.2, 50 MM (NH4)2SO4, AND 1 MM TCEP) WITH 0.5UL OF LIGAND (100 MM STOCK IN 100% DMSO) FOR 30 MINUTES AT 4C. CRYSTALS OF THE BINARY COMPLEX WERE OBTAINED BY MIXING 1.0UL OF THE PI3KG:INHIBITOR COMPLEX WITH 1.0UL OF A RESERVOIR SOLUTION CONSISTING OF 16-21% PEG 4K, 0.25 M (NH4)2SO4, AND 0.1 M TRIS-HCL, PH 7.5.