5LW6

Crystal structure of a Se-Met substituted Dictyostelium discoideum ADP-ribose binding macrodomain (residues 342-563) of DDB_G0293866

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	DIAMOND BEAMLINE I24
Synchrotron site	Diamond
Beamline	I24
Temperature [K]	100
Detector technology	PIXEL
Collection date	2013-05-23
Detector	DECTRIS PILATUS 6M-F
Wavelength(s)	0.97
Spacegroup name	P 21 21 21
Unit cell lengths	39.590, 71.220, 81.890
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	25.190 - 1.800
R-factor	0.17205
Rwork	0.170
R-free	0.21433
Structure solution method	SAD
RMSD bond length	0.020
RMSD bond angle	1.898
Data reduction software	XDS
Data scaling software	XDS
Phasing software	MLPHARE
Refinement software	REFMAC (5.7.0029)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	25.190	1.850
High resolution limit [Å]	1.800	1.800
Number of reflections	22050
<I/σ(I)>	17.3	5.3
Completeness [%]	99.9	100
Redundancy	25.8	25.4

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP		294	Crystallization trials were performed with proteins at 25 mg/ml in buffer containing 150 mM NaCl, 1 mM DTT and 25 mM Tris-HCl (pH 7.5), at 20 C with commercial screens using the sitting-drop vapour-diffusion method. Crystallization drops were set up with the aid of a Mosquito Crystal robot (TTP Labtech) using 200 nl of protein solution plus 200 nl of reservoir solution in MRC two-well crystallization microplates (Swissci) equilibrated against 75 microl of reservoir solution. Co-crystallisation trials were set up by adding 2 mM ADPr to the protein for at least 1 hour prior to setting up crystallisation drops. Crystals of the macrodomain proteins grew in 0.2 M Lithium sulphate, 0.1 M phosphate/citrate and 20% (w/v) PEG1000