6HMC

STRUCTURE OF PROTEIN KINASE CK2 CATALYTIC SUBUNIT (ISOFORM CK2ALPHA'; CSNK2A2 gene product) IN COMPLEX WITH THE INDENOINDOLE-TYPE INHIBITOR THN27

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE ID30B
Synchrotron site	ESRF
Beamline	ID30B
Temperature [K]	100
Detector technology	PIXEL
Collection date	2018-04-18
Detector	DECTRIS PILATUS3 S 6M
Wavelength(s)	0.8
Spacegroup name	P 1
Unit cell lengths	46.300, 47.300, 50.320
Unit cell angles	113.44, 90.38, 90.67

Refinement procedure

Resolution	46.293 - 1.030
R-factor	0.123
Rwork	0.123
R-free	0.13610
Structure solution method	AB INITIO PHASING
RMSD bond length	0.007
RMSD bond angle	1.002
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	Arcimboldo
Refinement software	PHENIX ((1.13_2998: ???))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	46.293	1.067
High resolution limit [Å]	1.030	1.030
Rmerge	0.052	0.919
Rmeas	0.062	1.089
Rpim	0.034	0.578
Number of reflections	179119	17408
<I/σ(I)>	9.22	1.02
Completeness [%]	92.3	90.48
Redundancy	3.4	3.5
CC(1/2)	0.997	0.562

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	8.5	293.15	180 MICROLITER ENZYME STOCK SOLUTION (6 MG/ML IN 500 MM NACL, 25 MM TRIS/HCL, PH 8.5) WERE MIXED and incubated WITH 20 MIKROLITER of a STOCK SOLUTION of the inhibitor 4B0 (10 MM 4B0 IN DMSO). 20 MICROLITERS OF the resulting solution WERE MIXED WITH 10 MICROLITERS OF RESERVOIR SOLUTION (900 MM LICL, 28 % (W/V) PEG 6000, 100 MM TRIS/HCL, PH 8.5) IN EACH WELL OF A CRYSTALLIZATION PLATE. A SINGLE MACROSEED, grown UNDER THE SAME CONDITIONS, WAS ADDED TO EACH DROPLET of the plate. THE DROPLETS WERE EQUILIBRATED AT 293.15 K using the sitting drop variant of the VAPOR DIFFUSION method. This procedure generated large single crystals of a CK2alpha'/4B0 complex. Afterwards, the inhibitor 4B0 was exchanged against THN27 by an extensive soaking procedure of several steps.