8SBC

Co-structure of Phosphatidylinositol 4,5-bisphosphate 3-kinase catalytic subunit alpha isoform and brain penetrant inhibitors

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ALS BEAMLINE 5.0.2
Synchrotron site	ALS
Beamline	5.0.2
Temperature [K]	100
Detector technology	PIXEL
Collection date	2016-01-21
Detector	DECTRIS PILATUS 6M
Wavelength(s)	1.000
Spacegroup name	P 21 21 21
Unit cell lengths	100.077, 107.847, 133.723
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	83.950 - 2.300
R-factor	0.2423
Rwork	0.241
R-free	0.27110
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.002
RMSD bond angle	0.386
Data reduction software	XDS
Data scaling software	Aimless
Phasing software	PHASER
Refinement software	PHENIX ((1.20.1_4487: ???))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	83.950	2.420
High resolution limit [Å]	2.300	2.300
Rmerge	0.075	1.522
Rmeas	0.079	1.590
Rpim	0.023	0.454
Total number of observations	823656	111247
Number of reflections	64894	9289
<I/σ(I)>	15.9	1.5
Completeness [%]	99.8
Redundancy	12.7	12
CC(1/2)	0.999	0.844

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7.2	303	RESERVOIR SOLUTION : 12%-20% PEG 3350, 0.125M-0.20M KSCN PROTEIN SOLUTION : 20MM TRIS PH 7.2, 200MM NACL, 1% BETAINE, 1% ETHYLENE GLYCOL, 0.02% CHAPS, 5MM DTT, PROTEIN IS 7.4 MG/ML FORMATION METHOD : SOAKING PROTOCOL : SET UP APO XTALS DROPS IN 1:1 RATIO IN 30C, REMARK 280 XTALS GROW OVERNIGHT, THEN ADD 2MM OF STOCK CPD AND INCUBATE FOR A FEW HOURS METHOD : VAPOR DIFFUSION - HANGING DROP TEMPERATURE : 303.0 CRYO PROTOCOL : 20% ETHYLENE GLYCOL PLUS WELL SOLUTION