3EYD

Structure of HCV NS3-4A Protease with an Inhibitor Derived from a Boronic Acid

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 17-ID
Synchrotron site	APS
Beamline	17-ID
Temperature [K]	100
Detector technology	CCD
Collection date	2004-04-07
Detector	ADSC QUANTUM 210
Wavelength(s)	1.000
Spacegroup name	H 3 2
Unit cell lengths	224.483, 224.483, 75.786
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	8.000 - 2.300
Rwork	0.180
R-free	0.25600
Structure solution method	FOURIER SYNTHESIS
Starting model (for MR)	HCV NS3(1-181)S139A - NS4a pdb2o8m.ent
RMSD bond length	0.009
RMSD bond angle	2.019
Data reduction software	HKL-2000
Data scaling software	SCALEPACK
Phasing software	X-PLOR
Refinement software	X-PLOR (98.1)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	50.000	2.200
High resolution limit [Å]	2.150	2.150
Rmerge	0.075	0.288
Number of reflections	28576
<I/σ(I)>	13.5	2
Completeness [%]	71.6	20.6
Redundancy	2.8

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	5.6		Crystallization was performed by the vapor diffusion method using hanging drops (4 L protein solution mixed with 4 L (0.75 - 1.00) M NaCl - 0.1 M MES - 0.1 M Na/K PO4, pH 5.6 - 6.2) suspended over 1 mL reservoir solutions of (1.25 - 1.50) M NaCl - 0.1 M MES - 0.1 M Na/K PO4 - 5 mM -mercaptoethanol, pH 5.6-6.2. The trays were set at 4oC for 5-7 days to control nucleation, followed by incubation for 3 weeks at 12oC to maximize crystal growth., VAPOR DIFFUSION, HANGING DROP, temperature 277, then 285K