4P1U

Influenza A (flu) virus polymerase basic protein 2 (PB2) bound to VX787, an azaindole inhibitor

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ALS BEAMLINE 5.0.1
Synchrotron site	ALS
Beamline	5.0.1
Temperature [K]	100
Detector technology	CCD
Collection date	2010-07-30
Detector	ADSC QUANTUM 210
Wavelength(s)	0.9774
Spacegroup name	P 65
Unit cell lengths	81.174, 81.174, 54.427
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	25.380 - 2.520
R-factor	0.1808
Rwork	0.178
R-free	0.24260
Structure solution method	FOURIER SYNTHESIS
RMSD bond length	0.010
RMSD bond angle	1.240
Refinement software	BUSTER

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	26.570	2.800
High resolution limit [Å]	2.520	2.520
Rmerge	0.045	0.388
Number of reflections	13202
<I/σ(I)>	22.8	3.4
Completeness [%]	97.3	98.8
Redundancy	4.3	4.3

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	4.7	293	1 ?L protein solution (2.8 mg/ml protein, 50 mM Tris buffer pH 8, 200 mM sodium chloride, 2 mM dithiothreitol, 1 mM anthraquinone-2,6-disulfonic acid disodium salt, 7.5 mM GTP) and 0.4 ?L well solution (1.5 M sodium formate, 100 mM sodium citrate buffer pH 4.7, 10 mM dithiothreitol) was suspended over 1 mL of well solution. The crystals were transferred to a soaking solution (3.25 M sodium formate, 100 mM sodium citrate buffer pH 4.7) containing 1 mM VRT-0928787. Crystals were incubated approximately 15 hours t room temperature, and then transferred to a cryo-preservative solution (soaking solution with 25 % v/v glycerol) prior to freezing in liquid nitrogen.