6GZM

Crystal Structure of Human CKIdelta with A86

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE MASSIF-1
Synchrotron site	ESRF
Beamline	MASSIF-1
Temperature [K]	110.15
Detector technology	PIXEL
Collection date	2016-12-19
Detector	DECTRIS PILATUS3 2M
Wavelength(s)	0.966
Spacegroup name	P 1 21 1
Unit cell lengths	49.232, 73.659, 89.204
Unit cell angles	90.00, 103.57, 90.00

Refinement procedure

Resolution	29.190 - 1.590
R-factor	0.19153
Rwork	0.189
R-free	0.24976
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	3uyt
RMSD bond length	0.012
RMSD bond angle	1.533
Data scaling software	SCALA
Refinement software	REFMAC (5.8.0158)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	29.190	1.680
High resolution limit [Å]	1.590	1.590
Rmeas	0.090	0.570
Number of reflections	80180	11624
<I/σ(I)>	7.1
Completeness [%]	96.5	96.4
Redundancy	2.4	2.4

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7.5	293.15	The construct used for crystallization was that of the wild type protein with the mutation R13N. Crystals of CK1d in complex with A-86 were obtained using hanging drop vapour diffusion set-ups. CK1d at a concentration of 13.6 mg/ml (50 mM HEPES, 266 mM NaCl, 1 mM EDTA, 1 mM DTT, 5 mM B-OG, pH 7.5) was pre-incubated with 2 mM (5.1-fold molar excess) of A-86 (150 mM in DMSO) for 1 h. 1 ul of the protein solution was then mixed with 1 ul of reservoir solution (0.1 M Na3-Citrate, pH 4.9, 18 %(w/v) PEG 3350) and equilibrated at 20 C over 0.4 ml of reservoir solution. Well diffracting crystals appeared over night