8SBJ

Co-structure Phosphatidylinositol 4,5-bisphosphate 3-kinase catalytic subunit alpha isoform complexed with 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	100.022, 107.750, 133.868
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	83.940 - 3.100
R-factor	0.2091
Rwork	0.207
R-free	0.25700
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.002
RMSD bond angle	0.396
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]	133.870	3.310
High resolution limit [Å]	3.100	3.100
Rmerge	0.154	1.485
Rmeas	0.161	1.543
Rpim	0.046	0.418
Total number of observations		63872
Number of reflections	26927	4789
<I/σ(I)>	15.3	2.1
Completeness [%]	99.9
Redundancy	12.2	13.3
CC(1/2)	0.999	0.809

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, XTALS GROW OVERNIGHT, THEN ADD 2MM OF STOCK CPD AND INCUBATE FOR A FEW HOURS; CRYO PROTOCOL: 20% ETHYLENE GLYCOL PLUS WELL SOLUTION