6H0U

Glycogen synthase kinase-3 beta (GSK3) complex with a covalent [1,2,4]triazolo[1,5-a][1,3,5]triazine inhibitor

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ELETTRA BEAMLINE 5.2R
Synchrotron site	ELETTRA
Beamline	5.2R
Temperature [K]	100
Detector technology	PIXEL
Collection date	2018-01-31
Detector	DECTRIS PILATUS 2M
Wavelength(s)	1.0
Spacegroup name	P 21 21 21
Unit cell lengths	86.803, 94.542, 106.579
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	47.270 - 2.300
R-factor	0.19483
Rwork	0.192
R-free	0.24047
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1q5k
RMSD bond length	0.009
RMSD bond angle	1.386
Data reduction software	XDS
Data scaling software	Aimless
Phasing software	MOLREP
Refinement software	REFMAC (5.8.0230)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	47.270	2.380
High resolution limit [Å]	2.300	2.300
Rpim	0.084	0.720
Number of reflections	39567	3843
<I/σ(I)>	7.4	1.5
Completeness [%]	99.8	100
Redundancy	3.7	3.8
CC(1/2)	0.991	0.570

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7	293	GSK3beta(35-386) aliquots were concentrated to 4.4 mg/ml and used for crystallization trials. GSK3b-inhibitor co-crystals were grown using the sitting drop vapor diffusion technique in 0.2M DL-Malic acid pH 7.0, 20% PEG 3350 as reservoir solution. The protein was previously incubated with 3x molar excess of compound for 3h at 4C. Crystallization drops were prepared from 0.5ul of protein solution and 0.5ul of reservoir, and incubated for 10 days at 20C. Crystals were cryoprotected in 30% Glycerol and frozen in liquid nitrogen prior to data collection.