7H6B

THE 2.17 A CRYSTAL STRUCTURE OF HUMAN CHYMASE IN COMPLEX WITH methyl 4-[(5-fluoro-3-methyl-1-benzothiophen-2-yl)sulfonylamino]-3-methylsulfonylbenzoate (TOA EIYO INHIBITOR)

This is a non-PDB format compatible entry.

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	BRUKER AXS MICROSTAR
Temperature [K]	100
Detector technology	IMAGE PLATE
Collection date	2004-11-24
Detector	MAR scanner 345 mm plate
Wavelength(s)	1.5418
Spacegroup name	P 21 21 21
Unit cell lengths	46.580, 53.520, 65.260
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	18.960 - 2.170
R-factor	0.2019
Rwork	0.200
R-free	0.23890
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.002
RMSD bond angle	0.513
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	PHASER
Refinement software	PHENIX ((1.21_5207: ???))

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	18.960	18.960	2.310
High resolution limit [Å]	2.180	6.200	2.180
Rmerge	0.077	0.028	0.404
Rmeas	0.088	0.033	0.465
Total number of observations	36488
Number of reflections	8882	389	1161
<I/σ(I)>	13.69	33.57	3.6
Completeness [%]	96.3

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	MICROBATCH	8.5	293	Sample of human Chymase in 50 mM MES/NaOH pH 5.5, 150mM NaCl, 1mM TCEP, 10% Glycerol) at a concentration of 11mg/ml to 14mg/ml.Add [2-[(4-methylpyridin-2-yl)amino]-2-oxoethyl] 2-methylquinoline-4-carboxylate at 10x molar ratio. The compound helps to obtain crystals but is not visible in the structures. Add 0.5 mM ZnCl2. Add inhibitor, incubate for 16h on ice. Crystallize using microbatch setups with Al's oil (Hampton Research), with total drop size 1ul with 50% protein sample, using crystallization reagent of 0.1M Tris/HCl pH 8.5, 0.2M NaCl, 25% PEG 3350.