5CE1

Crystal Structure of Serine protease Hepsin in complex with Inhibitor

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	RIGAKU RU300
Temperature [K]	100
Detector technology	IMAGE PLATE
Collection date	2012-02-12
Detector	RIGAKU RAXIS IV++
Wavelength(s)	1.5
Spacegroup name	P 1 21 1
Unit cell lengths	59.583, 47.678, 60.784
Unit cell angles	90.00, 105.30, 90.00

Refinement procedure

Resolution	58.630 - 2.500
R-factor	0.19165
Rwork	0.187
R-free	0.28150
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1z8g
RMSD bond length	0.014
RMSD bond angle	1.695
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	MOLREP
Refinement software	REFMAC (5.7.0029)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	100.000	2.590
High resolution limit [Å]	2.500	2.500
Rmerge	0.074	0.229
Number of reflections	10137
<I/σ(I)>	2.4
Completeness [%]	87.4	89.9
Redundancy	2.6	2.2

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	6	293	Activated hepsin was incubated with 10 mM Benzamidine at 4 oC for 2 hours and concentrated to7 mg/ml using 10 KDa cut off centricon [Millipore]. 1microlit of protein was mixed with 1 microlit of reservoir buffer containing 100 mM sodium Cacodylate (6.0), 22% PEG 3350, and 200 mM ammonium fluoride. Plate clusters with a dimension of 50 micron m x 100 micron m were observed after 2 days of crystallization set up.