7H68

THE 1.25 A CRYSTAL STRUCTURE OF HUMAN CHYMASE IN COMPLEX WITH (2R)-2-benzyl-4-[(R)-(5-fluoro-3-methyl-1H-indol-2-yl)-phenylmethyl]-3-hydroxy-2H-furan-5-one

This is a non-PDB format compatible entry.

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SLS BEAMLINE X06SA
Synchrotron site	SLS
Beamline	X06SA
Temperature [K]	100
Detector technology	CCD
Collection date	2009-03-11
Detector	MARMOSAIC 225 mm CCD
Wavelength(s)	1
Spacegroup name	P 43
Unit cell lengths	73.600, 73.600, 48.484
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	32.910 - 1.250
R-factor	0.12605
Rwork	0.125
R-free	0.14810
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.009
RMSD bond angle	1.439
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	PHASER
Refinement software	REFMAC (5.7.0029)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	32.910	32.910	1.250
High resolution limit [Å]	1.220	5.440	1.220
Rmerge	0.031	0.020	0.457
Rmeas	0.036	0.024	0.611
Total number of observations	213822
Number of reflections	67565	823	1872
<I/σ(I)>	19.65	49.47	1.38
Completeness [%]	86.6
CC(1/2)	0.999	0.999	0.612

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.