4E7A

Crystal structure of a product state assembly of HCV NS5B genotype 2a JFH-1 isolate with beta hairpin deletion bound to primer-template RNA with a 2',3'-ddC

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 21-ID-G
Synchrotron site	APS
Beamline	21-ID-G
Temperature [K]	100
Detector technology	CCD
Collection date	2011-12-16
Detector	MARMOSAIC 300 mm CCD
Wavelength(s)	0.97856
Spacegroup name	P 65
Unit cell lengths	142.730, 142.730, 91.500
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	50.000 - 3.000
R-factor	0.197
Rwork	0.194
R-free	0.24100
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4e76
RMSD bond length	0.009
RMSD bond angle	1.340
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	PHASER
Refinement software	REFMAC

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	50.000		3.080
High resolution limit [Å]	3.000	13.420	3.000
Rmerge	0.099	0.020	0.745
Number of reflections	21289	235	1572
<I/σ(I)>	16.76	54.99	2.27
Completeness [%]	99.5	89.7	99.6
Redundancy	5.3

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	6	289	4.37 mg/mL NS5B in 20 mM Tris, pH 8, 200 mM sodium chloride, 20% glycerol, 2 mM TCEP, 200 mM imidazole against 30% PEG3350, 0.1 M Bis-Tris propane, pH 6.0, 200 mM ammonium acetate soaked overnight in precipitant supplemented with 15% ethylene glycol as cryoprotectant and 0.2 mM CUAGGC(DOC), crystal tracking ID 228018H11, VAPOR DIFFUSION, SITTING DROP, temperature 289K