4OBC

Crystal structure of HCV polymerase NS5b genotype 2a JFH-1 isolate with the S15G, C223H, V321I resistance mutations against the guanosine analog GS-0938 (PSI-3529238)

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SSRL BEAMLINE BL9-2
Synchrotron site	SSRL
Beamline	BL9-2
Temperature [K]	100
Detector technology	CCD
Collection date	2011-06-30
Detector	MARMOSAIC 325 mm CCD
Wavelength(s)	0.97946
Spacegroup name	P 65
Unit cell lengths	140.208, 140.208, 92.567
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	50.000 - 2.500
R-factor	0.1899
Rwork	0.188
R-free	0.22340
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1yuy
RMSD bond length	0.013
RMSD bond angle	1.421
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	PHASER (2.1.4)
Refinement software	REFMAC (5.5.0109)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	50.000	50.000	2.570
High resolution limit [Å]	2.500	5.720	2.500
Rmerge	0.085	0.046	0.638
Number of reflections	35841
<I/σ(I)>	21.9		2.3
Completeness [%]	99.7	99.7	97.3
Redundancy	7.8	8.8	3.7

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	8	289	NS5b VCID 5854 at 4.25 mg/mL in 5 mM Tris pH 7.5, 200 mM NH4OAc, 1 mM EDTA 1 mM DTT against PACT screen condition D5, 25% PEG 1500, 0.1M MMT Malic Acid, MES, Tris Buffer pH 8.0 supplemented with 20% glycerol as cryo-protectant, crystal tracking ID 223572d5, puck ID cps0237-1, VAPOR DIFFUSION, SITTING DROP, temperature 289K