2YI9

Structure of the RNA polymerase VP1 from Infectious Pancreatic Necrosis Virus in complex with magnesium

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	DIAMOND BEAMLINE I02
Synchrotron site	Diamond
Beamline	I02
Temperature [K]	100
Detector technology	CCD
Collection date	2009-06-29
Detector	ADSC CCD
Spacegroup name	P 21 21 21
Unit cell lengths	134.048, 183.874, 244.521
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	32.210 - 2.200
R-factor	0.1605
Rwork	0.160
R-free	0.18120
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2pgg
RMSD bond length	0.011
RMSD bond angle	0.980
Data reduction software	MOSFLM
Data scaling software	SCALA
Phasing software	PHASER
Refinement software	BUSTER (2.9.2)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	101.810	2.260
High resolution limit [Å]	2.200	2.200
Rmerge	0.170	0.750
Number of reflections	304685
<I/σ(I)>	10.6	2.7
Completeness [%]	100.0	100
Redundancy	7.3	7.4

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP		293.65	CRYSTALS WERE GROWN IN SITTING DROPS CONTAINING 200 NL PROTEIN (5.8-6.8 MG/ML) AND 100 NL RESERVOIR SOLUTION (20-18% W/V PEG 3350, 0.10-0.09 M BIS-TRIS PROPANE PH 7.5, 0.2-0.18 M SODIUM CITRATE) EQUILIBRATED AGAINST 95 UL RESERVOIRS AT 20.5 DEGREES CELSIUS. CRYSTALS WERE SOAKED BY DILUTING THE MOTHER-LIQUOR WITH APPROX. 0.5 UL 20% W/V PEG 3350, 0.1 M BIS-TRIS PROPANE PH 8.25 AND 20% V/V GLYCEROL, TRANSFERRING THE CRYSTALS TO A FRESH DROP CONTAINING 20% W/V PEG 3350, 0.1 M BIS-TRIS PROPANE PH 8.25, 20% V/V GLYCEROL, 50 MM MGCL2 AND 10 UM GTP, AND INCUBATING FOR 10 MIN.