4WT9

APO CRYSTAL STRUCTURE OF HCV NS5B GENOTYPE 2A JFH-1 ISOLATE WITH E86Q E87Q S15G C223H V321I AND DELTA8 MUTATIONS

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 21-ID-D
Synchrotron site	APS
Beamline	21-ID-D
Temperature [K]	100
Detector technology	CCD
Collection date	2012-03-28
Detector	MARMOSAIC 300 mm CCD
Wavelength(s)	1.07809
Spacegroup name	P 65
Unit cell lengths	140.370, 140.370, 92.230
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	50.000 - 2.500
R-factor	0.195
Rwork	0.193
R-free	0.24500
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4e76
RMSD bond length	0.012
RMSD bond angle	1.529
Data scaling software	XSCALE
Phasing software	PHASER
Refinement software	REFMAC (5.6.0117)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	50.000	2.560
High resolution limit [Å]	2.500	2.500
Rmerge	0.084	0.706
Rmeas	0.099	0.021
Total number of observations	127140
Number of reflections	35345	394
<I/σ(I)>	16.62	2.6
Completeness [%]	98.5	98.6
Redundancy	3.6	3.7

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	6.9	289	NS5B AT 4.08 MG/ML IN 20 MM TRIS PH 8, 200 MM NACL, 20% GLYCEROL, 2 MM TCEP, 200 MM IMIDAZOLE AGAINST 30% PEG 550 MME, 0.1 M HEPES PH 7.5, 50 MM MAGNESIUM CHLORIDE WITH 15% ETHYLENE GLYCOL AS CRYO-PROTECTANT, CRYSTAL TRACKING ID 232769E7