5N9N

Crystal structure of human Protein kinase CK2 catalytic subunit in complex with the ATP-competitive, tight-binding dibenzofuran inhibitor TF85 (4a)

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SLS BEAMLINE X06DA
Synchrotron site	SLS
Beamline	X06DA
Temperature [K]	100
Detector technology	PIXEL
Collection date	2012-10-12
Detector	DECTRIS PILATUS 2M
Wavelength(s)	0.99987
Spacegroup name	P 1 21 1
Unit cell lengths	57.477, 45.505, 63.489
Unit cell angles	90.00, 110.97, 90.00

Refinement procedure

Resolution	34.165 - 1.841
R-factor	0.1686
Rwork	0.167
R-free	0.20110
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2pvr
RMSD bond length	0.003
RMSD bond angle	0.568
Data reduction software	XDS
Data scaling software	Aimless
Phasing software	PHASER
Refinement software	PHENIX ((1.11.1_2575: ???))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	45.510	1.880
High resolution limit [Å]	1.840	1.840
Rmerge	0.056	0.750
Number of reflections	25795	1395
<I/σ(I)>	12.6
Completeness [%]	96.4	84.2
Redundancy	3.5	3
CC(1/2)	0.998	0.644

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	5.6	293	Prior to the crystallization the Inhibitor TF85 was solubilized in 100 % DMSO in a concentration of 10 mM. This TF85 stock solution was mixed with human CK2alpha (construct 1-335; Protein concentration 8-10 mg/ml in 500 mM sodium chloride, 25 mM Tris/HCl pH 8.5) in a ratio of 1:10. After a short time of incubation, this mixture was mixed with reservoir solution [32 % (w/v) PEG4000, 0.2 M ammonium acetate, 0.1 M citrate pH 5.6] in a ratio of 5:2. 3.5 microliter of these mixtures were then equilibrated against the reservoir solution. The crystal growth was induced by seeding with 150 nanoliter seeding suspension after an equilibration time of two days.