7UY0

Crystal structure of human Fgr tyrosine kinase in complex with A-419259

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 23-ID-D
Synchrotron site	APS
Beamline	23-ID-D
Temperature [K]	100
Detector technology	PIXEL
Collection date	2018-11-09
Detector	DECTRIS PILATUS3 6M
Wavelength(s)	1.03326
Spacegroup name	P 21 21 21
Unit cell lengths	49.430, 112.090, 213.730
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	77.346 - 2.550
R-factor	0.1883
Rwork	0.186
R-free	0.22290
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2src
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	PHASER
Refinement software	PHENIX (1.14_3260)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	77.346	77.346	2.620
High resolution limit [Å]	2.550	11.400	2.550
Rmerge	0.120	0.056	1.180
Rmeas	0.125	0.059	1.227
Total number of observations	501528
Number of reflections	39772	531	2917
<I/σ(I)>	13.4	32.69	2.07
Completeness [%]	100.0	99.6	100
Redundancy	12.61	10.77	12.909
CC(1/2)	0.998	0.997	0.770

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION	7.5	277	For co-crystallization with A-419259, Fgr (3.2 mg/mL in 20 mM Tris-HCl, pH 8.3, 100 mM NaCl, and 2 mM TCEP) was mixed with 10 mM A-419259 (in 50% DMSO) to a final inhibitor concentration of 120 microM (0.3% DMSO final) and incubated for 30 min at 298 K prior to crystallization setup. Crystals were grown by sitting- and hanging-drop vapor diffusion at 277 K by mixing Fgr/A-419259 in a 1:1 ratio with the mother liquor (0.2 M sodium/potassium phosphate, 0.1 M Bis-Tris propane, pH 7.5, and 16% PEG 3350). To promote crystal growth, crystal seeds from initial needle-like crystals were added to the crystallization drops