7P9N

Crystal structure of CD73 in complex with AMP in the open form

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	BESSY BEAMLINE 14.1
Synchrotron site	BESSY
Beamline	14.1
Temperature [K]	100
Detector technology	PIXEL
Collection date	2016-06-23
Detector	DECTRIS PILATUS 6M
Wavelength(s)	0.91841
Spacegroup name	P 21 21 2
Unit cell lengths	67.345, 131.346, 66.253
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	47.270 - 1.550
R-factor	0.1533
Rwork	0.152
R-free	0.17720
Structure solution method	FOURIER SYNTHESIS
Starting model (for MR)	4h2g
RMSD bond length	0.015
RMSD bond angle	2.041
Data reduction software	XDS
Data scaling software	Aimless (0.7.4)
Phasing software	REFMAC
Refinement software	REFMAC (5.8.0267)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	47.270	47.230	1.580
High resolution limit [Å]	1.550	8.510	1.550
Rmerge	0.103	0.048	0.422
Rmeas	0.113	0.053	0.487
Rpim	0.044	0.022	0.235
Total number of observations		3311	9598
Number of reflections	78906	602	2429
<I/σ(I)>	11.5	26.1	2.6
Completeness [%]	92.6	98.4	58.5
Redundancy	6.2	5.5	4
CC(1/2)	0.997	0.997	0.822

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7.8	292.15	7 mg/mL protein concentration, 100 mM Tris pH 7.8, 10 % PEG6000, equal amounts of protein and reservoir. Following crystal formation (1-2 days), the crystals were transferred to soaking solution containing reservoir solution and 100 mM AMP. Crystals were then transferred to cryo solution containing an additional 20 % glycerol, soaked for ~2-5 min, and flash frozen in liquid nitrogen.