6HME

LOW-SALT STRUCTURE OF PROTEIN KINASE CK2 CATALYTIC SUBUNIT (ISOFORM CK2ALPHA; CSNK2A1 gene product) IN COMPLEX WITH THE INDENOINDOLE-TYPE INHIBITOR THN27

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	PETRA III, EMBL c/o DESY BEAMLINE P13 (MX1)
Synchrotron site	PETRA III, EMBL c/o DESY
Beamline	P13 (MX1)
Temperature [K]	100
Detector technology	PIXEL
Collection date	2018-06-18
Detector	DECTRIS PILATUS 6M
Wavelength(s)	0.9794
Spacegroup name	P 43 21 2
Unit cell lengths	126.370, 126.370, 124.630
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	89.357 - 1.850
R-factor	0.1754
Rwork	0.175
R-free	0.19700
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	5oni
RMSD bond length	0.006
RMSD bond angle	0.835
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]	89.357	1.916
High resolution limit [Å]	1.850	1.850
Rmerge	0.055	1.701
Rmeas	0.060	1.843
Rpim	0.023	0.705
Number of reflections	86172	8521
<I/σ(I)>	16.12	0.96
Completeness [%]	99.8	99.88
Redundancy	6.5	6.8
CC(1/2)	0.999	0.535

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	6.5	293.15	180 MICROLITERS OF ENZYME SOLUTION (6 MG/ML CK2ALPHA, 0.025 M TRIS/HCL, PH 8.5, 0.5 M NACL) WERE MIXED WITH 20 MICROLITERS OF INHIBITOR STOCK SOLUTION (0.010 M INHIBITOR THN27 IN DMSO). THIS MIXTURE WAS INCUBATED FOR 30 MIN AT ROOM TEMPERATURE. THE RESERVOIR SOLUTION OF THE CRYSTALLIZATION EXPERIMENT WAS 0.2 M AMMONIUM SULFATE, 0.1 M SODIUM CACODYLATE TRIHYDRATE, PH 6.5, 30% (W/V) PEG 8,000. PRIOR TO EQUILIBRATION THE CRYSTALLIZATION DROP WAS COMPOSED OF 10 MICROLITERS RESERVOIR SOLUTION PLUS 20 MICROLITERS ENZYME/INHIBITOR MIXTURE.