7H67

THE 1.35 A CRYSTAL STRUCTURE OF HUMAN CHYMASE IN COMPLEX WITH 2-[(3,5-dimethyl-1-benzothiophen-2-yl)-phenylmethyl]-3-hydroxy-4-methyl-4-phenylcyclobut-2-en-1-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	2006-09-27
Detector	MARMOSAIC 225 mm CCD
Wavelength(s)	0.99985
Spacegroup name	P 43
Unit cell lengths	73.800, 73.800, 48.850
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	73.720 - 1.350
R-factor	0.15861
Rwork	0.158
R-free	0.17551
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.011
RMSD bond angle	1.698
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	PHASER
Refinement software	REFMAC (5.3.0008)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	73.720	73.720	1.430
High resolution limit [Å]	1.350	4.000	1.350
Rmerge	0.047	0.024	0.282
Rmeas	0.054	0.028	0.363
Total number of observations	241878
Number of reflections	56943	2224	8357
<I/σ(I)>	21.14	40.23	5.65
Completeness [%]	98.5

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.