1C5X

STRUCTURAL BASIS FOR SELECTIVITY OF A SMALL MOLECULE, S1-BINDING, SUB-MICROMOLAR INHIBITOR OF UROKINASE TYPE PLASMINOGEN ACTIVATOR

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Temperature [K]	298
Detector technology	IMAGE PLATE
Collection date	1998-03-22
Detector	RIGAKU RAXIS IV++
Spacegroup name	C 1 2 1
Unit cell lengths	82.090, 49.780, 66.650
Unit cell angles	90.00, 113.29, 90.00

Refinement procedure

Resolution	7.500 - 1.750
R-factor	0.202 *
Rwork	0.195
R-free	0.24400
Structure solution method	DIFFERENCE FOURIER PLUS REFINEMENT
Starting model (for MR)	PDB1LMW
RMSD bond length	0.017
RMSD bond angle	4.000
Data reduction software	bioteX ((MSC))
Data scaling software	bioteX
Phasing software	X-PLOR (3.1)
Refinement software	X-PLOR (3.1)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	38.490	1.830
High resolution limit [Å]	1.320	1.750
Rmerge	0.088	0.235
<I/σ(I)>	4	1.6
Completeness [%]	68.0	38.9
Redundancy	2.8

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	Vapor diffusion, hanging drop *	7.4 *		LMW human uPA/A145 was concentrated to 10 mg/ml and incubated in 50 mM HEPES, 5.0 mM NaCl. pH 7.4, 1.4 mM 4-iodobenzo[b]thiophene-2-carboxamidine for 15 min on ice. The complex was crystallized by vapor diffusion in hanging drops containing equal volumes of protein-inhibitor solution (0.28 mM uPA/A145, 1.4 mM inhibitor) and well solution (20 % 2-propanol, 20 % PEG 4K, 100 mM sodium citrate, pH 6.5) sealed over the well.

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	drop	protein	0.28 (mM)
2	1	drop	inhibitor	1.4 (mM)
3	1	reservoir	2-propanol	20 (%)
4	1	reservoir	PEG4000	20 (%)
5	1	reservoir	sodium citrate	100 (mM)