7PD9

Crystal structure of CD73 in complex with riboflavin 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	CCD
Collection date	2016-10-04
Detector	RAYONIX MX-225
Wavelength(s)	0.89429
Spacegroup name	P 21 21 2
Unit cell lengths	67.396, 131.782, 66.517
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	47.160 - 1.390
R-factor	0.1216
Rwork	0.120
R-free	0.14940
Structure solution method	FOURIER SYNTHESIS
Starting model (for MR)	4h2g
RMSD bond length	0.016
RMSD bond angle	1.966
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]	47.160	47.120	1.420
High resolution limit [Å]	1.390	7.620	1.390
Rmerge	0.059	0.017	0.751
Rmeas	0.066	0.020	0.838
Rpim	0.029	0.009	0.369
Total number of observations		3375	27824
Number of reflections	117101	833	5581
<I/σ(I)>	13.8	41.7	1.7
Completeness [%]	98.4	98.3	95.7
Redundancy	5	4.1	5
CC(1/2)	0.999	1.000	0.700

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP		292	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 6.25 mM riboflavin. Crystals were then transferred to cryo solution containing an additional 20 % glycerol, soaked for ~2-5 min, and flash frozen in liquid nitrogen.