6RUU

Pseudokinase domain of human IRAK3

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	DIAMOND BEAMLINE I24
Synchrotron site	Diamond
Beamline	I24
Temperature [K]	100
Detector technology	PIXEL
Collection date	2018-10-14
Detector	DECTRIS PILATUS3 6M
Wavelength(s)	1.007
Spacegroup name	P 21 21 2
Unit cell lengths	53.550, 167.160, 179.510
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	48.650 - 2.950
R-factor	0.2299
Rwork	0.228
R-free	0.26000
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2qkw
RMSD bond length	0.003
RMSD bond angle	0.627
Data reduction software	XDS (20190315)
Data scaling software	Aimless (0.7.4)
Phasing software	MoRDa (ccp4-online)
Refinement software	PHENIX

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	49.056	3.307
High resolution limit [Å]	2.500	2.959
Rmerge	0.280	2.400
Rmeas	0.285	2.450
Rpim	0.052	0.438
Number of reflections	28005	1072
<I/σ(I)>	12.1	1.8
Completeness [%]	92.2	59.6
Redundancy	29.4	30.9
CC(1/2)	0.998	0.702

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	5	293.15	20C, 8mg/mL protein, hanging drop vapour diffusion, 2uL :1uL drop ratio (protein:precipitant) + 1mL reservoir, streak seeding from 1:100 seed stock (original seed stock: 10 uL from drops containing previously obtained crystals +50 uL precipitant mix, crushed by vortexing with seed bead, diluted in precipitant mix), crystals appeared after 2-3 days, harvested after 5-7 days, overnight soaking with 1:1 v/v of 5 mM Ethylmercury Phosphate in stabilising solution, Cryo: Precipitant mix +30% Glycerol