8SSN

Abl kinase in complex with SKI and asciminib

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ALS BEAMLINE 2.0.1
Synchrotron site	ALS
Beamline	2.0.1
Temperature [K]	100
Detector technology	PIXEL
Collection date	2022-06-01
Detector	DECTRIS PILATUS 6M
Wavelength(s)	1.04054
Spacegroup name	P 21 21 21
Unit cell lengths	92.478, 103.213, 107.963
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	47.830 - 2.860
R-factor	0.3002
Rwork	0.298
R-free	0.34800
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.004
RMSD bond angle	0.729
Data reduction software	XDS
Data scaling software	Aimless
Phasing software	PHASER
Refinement software	PHENIX ((1.20.1_4487: ???))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	47.830	2.967
High resolution limit [Å]	2.860	2.860
Rmerge	0.086	1.367
Number of reflections	24343	2372
<I/σ(I)>	4.93	0.51
Completeness [%]	98.3
Redundancy	2
CC(1/2)	0.997	0.310

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	8	291.15	Crystals of AblFL in complex with SKI and asciminib were obtained by combining 0.3 ul of 600 uM AblFL + 700 uM SKI + 700 uM asciminib (~32 mg/ml) in 5 percent DMSO with 0.4 ul reservoir of 0.1 M Tris-HCl pH 8 + 1.75 M Ammonium sulfate + 2 percent (v/v) polypropylene glycol 400 (PPG 400). The final stock of complex was concentrated from 1 uM AblFL with ~1.2 uM SKI/asciminib after incubation at 4 degree C for 6 h. Screening around this condition yielded crystals in a transparent diamond-shaped or plate-shaped crystals. Crystals were grown at 18 degree C by sitting drop for a few days. The crystals were transferred to a drop of fresh reservoir containing 20 percent Xylitol with matching concentration of inhibitors in 5 percent DMSO for few seconds for cryo-protection