5JBJ

Crystal structure of chicken LGP2 with 5'p 12-mer dsRNA at 3.6 A resolution

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE ID23-1
Synchrotron site	ESRF
Beamline	ID23-1
Temperature [K]	100
Detector technology	PIXEL
Collection date	2014-02-24
Detector	DECTRIS PILATUS3 6M
Wavelength(s)	0.9786
Spacegroup name	P 64
Unit cell lengths	90.090, 90.090, 196.730
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	78.020 - 3.580
R-factor	0.25647
Rwork	0.253
R-free	0.31881
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	5jb2
RMSD bond length	0.006
RMSD bond angle	0.954
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	PHASER
Refinement software	REFMAC (5.8.0135)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	78.020	3.730
High resolution limit [Å]	3.580	3.580
Rmerge	0.067	1.180
Number of reflections	10663
<I/σ(I)>	20.74
Completeness [%]	78.0	99
Redundancy	11.51	10.32

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	6.5	277	chLGP2, directly after size exclusion chromatography, was mixed with dsRNA in a 1:1 ratio and incubated for 30 minutes on ice. The complex was concentrated with an Amicon Ultra concentrator to around 10 mg/ml and then 2 mM ADP:AlF4 and 2 mM MgCl2 were added. chLGP2:12bp dsRNA:ADP:AlF4 complex was mixed with reservoir buffer (0.1 M Bis-Tris propane pH 6.5, 0.05-0.1 M magnesium formate) in a 2:1 ratio