2P3U

Crystal structure of human factor XA complexed with 3-chloro-N-(4-chloro-2-{[(5-chloropyridin-2-yl)amino]carbonyl}-6-methoxyphenyl)-4-[(1-methyl-1H-imidazol-2-yl)methyl]thiophene-2-carboxamide {Pfizer 320663}

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SSRL BEAMLINE BL7-1
Synchrotron site	SSRL
Beamline	BL7-1
Temperature [K]	100
Detector technology	IMAGE PLATE
Collection date	2000-03-11
Detector	MAR scanner 345 mm plate
Spacegroup name	P 21 21 21
Unit cell lengths	56.014, 71.717, 78.651
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	19.800 - 1.620
R-factor	0.192
Rwork	0.191
R-free	0.21000
Structure solution method	OTHER
Starting model (for MR)	1fjs
RMSD bond length	0.004
RMSD bond angle	0.900
Data reduction software	MOSFLM
Data scaling software	SCALA
Phasing software	X-PLOR
Refinement software	CNX

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	20.000	1.660
High resolution limit [Å]	1.620	1.620
Number of reflections	40439
<I/σ(I)>	18.4	3.2
Completeness [%]	98.9	97.9
Redundancy	3.7	3.7

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	EVAPORATION	7.5	295	A THREE-FOLD EXCESS OF 3-Chloro-4-(2-methylamino-imidazol-1-ylmethyl)-thiophene-2-carboxylic acid [4-chloro-2-(5-chloro-pyridin-2-ylcarbamoyl)-6-methoxy-phenyl]-amide WAS ADDED TO THE DES-GLA-FACTOR XA. THE PROTEIN WAS THEN CONCENTRATED TO 12-17 MG/ML. CRYSTALS WERE GROWN USING 2 UL OF COMPLEX WITH 2 UL OF RESERVOIR CONTAINING 15-21% PEG1500 AND 10 MM CACL2. 30-40 UL SITTING DROPS CONTAINING SATURATED INHIBITOR (5 MM) IN 21% PEG1500, 5 MM CACL2, 20 MM NACL, 25 MM TRIS PH 7.5 (CRYSTAL SOAKING SOLUTION) WERE EQUILIBRATED OVER A 1 ML RESERVOIR CONTAINING THE CRYSTAL SOAKING SOLUTION FOR 1 TO 2 DAYS. A SINGLE FACTOR XA CRYSTAL WAS TRANSFERRED USING A MOUNTED CRYOLOOP INTO ONE OF THESE SITTING DROPS AND ALLOWED TO SOAK FOR THREE OR MORE DAYS, EVAPORATION, temperature 295K