5BUH

Influenza PB2 bound to a hydroxymethyl azaindole inhibitor

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ALS BEAMLINE 5.0.2
Synchrotron site	ALS
Beamline	5.0.2
Temperature [K]	100
Detector technology	CCD
Collection date	2010-04-21
Detector	ADSC QUANTUM 315
Wavelength(s)	1.0
Spacegroup name	P 65
Unit cell lengths	81.801, 81.801, 55.598
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	26.780 - 2.550
R-factor	0.1716
Rwork	0.170
R-free	0.19830
Structure solution method	FOURIER SYNTHESIS
RMSD bond length	0.010
RMSD bond angle	1.110
Data scaling software	autoPROC
Refinement software	BUSTER-TNT

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	26.780	26.780	2.850
High resolution limit [Å]	2.550	8.060	2.550
Rmerge	0.051	0.025	0.386
Total number of observations	29301	921	4101
Number of reflections	6918
<I/σ(I)>	17.1
Completeness [%]	98.5	96.9	98
Redundancy	4.2	4.1	4.2

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	4.7	293	1 microL 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 small-molecule inhibitor. 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.