8QQ6

CRYSTAL STRUCTURE OF FVIIA IN COMPLEX WITH A BENZAMIDINE-BASED INHIBITOR

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE ID14-2
Synchrotron site	ESRF
Beamline	ID14-2
Temperature [K]	100
Detector technology	CCD
Collection date	2004-07-09
Detector	ADSC QUANTUM 4
Wavelength(s)	0.933
Spacegroup name	P 21 21 21
Unit cell lengths	70.537, 82.605, 126.367
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	19.940 - 1.870
R-factor	0.1953
Rwork	0.192
R-free	0.25920
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.008
RMSD bond angle	1.010
Data reduction software	XDS
Data scaling software	STARANISO
Phasing software	PHASER
Refinement software	BUSTER (2.11.8)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	61.591	2.119
High resolution limit [Å]	1.870	1.870
Rmerge	0.076	0.826
Rmeas		0.939
Rpim		0.439
Number of reflections	34451	1725
<I/σ(I)>	15.5	1.7
Completeness [%]	55.8	9
Redundancy	4.7	4.3
CC(1/2)		0.724

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	6.5	293	The VIIa/sTF complex was crystallized using the sitting drop or the hanging drop vapor diffusion method. Specifically, the protein drop contained 4 mg/ml VIIa/sTF complex, 20 mm Tris-HCl, pH 7.5, 200 mm NaCl, 10 mm CaCl2, and 10 mm pAB or 10 mm benzamidine, whereas the reservoir solution contained 16-22% PEG 4000, 100 mm MgCl2, and 20 mm BisTris, pH 6.5. Drops were prepared by mixing 2 ul of protein solution with 2 ul of reservoir solution at 20 C. Crystals appeared within 7 days and were allowed to grow up to 14-20 days before being flash-frozen without additional cryoprotectant.