8UM5

X-ray structure of human SHIP1 Ptase-C2 domains covalently bound to TREAT-AD (TAD) compound TAD-58547

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 21-ID-F
Synchrotron site	APS
Beamline	21-ID-F
Temperature [K]	100
Detector technology	CCD
Collection date	2021-10-01
Detector	RAYONIX MX-300
Wavelength(s)	0.97872
Spacegroup name	P 1 21 1
Unit cell lengths	48.643, 82.266, 59.749
Unit cell angles	90.00, 108.55, 90.00

Refinement procedure

Resolution	24.870 - 1.860
R-factor	0.1602
Rwork	0.158
R-free	0.20770
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.009
RMSD bond angle	0.970
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	PHENIX
Refinement software	PHENIX (1.20.1_4487)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	24.870	1.927
High resolution limit [Å]	1.860	1.861
Rmerge	0.065	0.614
Rmeas	0.073	0.689
Rpim	0.032	0.310
Number of reflections	36968	16154
<I/σ(I)>	15.99	2.49
Completeness [%]	98.3	90.79
Redundancy	4.9	4.7
CC(1/2)	0.998	0.831

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP		293	17-22% PEG 2000 MME, 7-10% PEG 3000 and 0.1M HEPES pH 7.0. Protein crystallization was performed in 24-well sitting drop plates with a 1:1 ratio of protein to reservoir solution (final volume of 3 uL). Crystals were allowed to grow for at least 1 week at room temperature. Crystals at sizes appropriate for X-ray data collection were then cryoprotected with 2.5 uL of cryosolution (90% reservoir solution + 10% PEG 400.) Crystals were harvested with nylon loops and flash-frozen in liquid nitrogen.