7B8I

Tetragonal structure of human protein kinase CK2 catalytic subunit in complex with a heparin oligo saccharide

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	2014-06-05
Detector	DECTRIS PILATUS 2M-F
Wavelength(s)	0.97795
Spacegroup name	P 43 21 2
Unit cell lengths	128.401, 128.401, 124.154
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	45.397 - 2.550
R-factor	0.2057
Rwork	0.204
R-free	0.23960
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2pvr
RMSD bond length	0.002
RMSD bond angle	0.534
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	PHASER
Refinement software	PHENIX ((1.13_2998: ???))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	45.400	2.642
High resolution limit [Å]	2.550	2.550
Rmerge	0.085	0.723
Rmeas	0.093	0.788
Rpim	0.036	0.307
Number of reflections	33919	3259
<I/σ(I)>	16.16	2.57
Completeness [%]	98.4	96.19
Redundancy	6.7	6.4
CC(1/2)	0.999	0.802

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP		293.15	Prior to crystallization, the enzyme was incubated with heparin decasaccharide; the composition of this preincubation solution was 5 mg/ml CK2alpha1-335, 1.4 mM Heparin decasaccharide, 340 mM NaCl, 25 mM Tris/HCl, pH 8.5. 4 microliter of this enzyme/heparin mixture was mixed with 1 microliter reservoir solution. The composition of the reservoir solution was 32 % (w/v) PEG4000, 0.2 M Lithium sulfate, 0.1 M Tris/HCl, pH 7.5. As a preparation of X-ray diffraction data collection, the crystals were transferred to a cryo solution composed of 32 % (w/v) PEG4000, 10 % (v/v) glycerol, 0.2 M lithium sulfate, 0.5 mM Heparin decasaccharide, 0.1 M Tris/HCl, pH 7.5.