7H6F

THE 1.25 A CRYSTAL STRUCTURE OF HUMAN CHYMASE IN COMPLEX WITH 3-[2-(dimethylamino)-2-oxoethyl]-1-[(5-fluoro-1-benzothiophen-3-yl)methyl]indole-2-carboxylic acid (INDOLE 2-CARBOXYLIC ACID)

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	2006-02-20
Detector	MARMOSAIC 225 mm CCD
Wavelength(s)	1.003328
Spacegroup name	P 43
Unit cell lengths	71.870, 71.870, 47.950
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	19.950 - 1.250
R-factor	0.14585
Rwork	0.144
R-free	0.17195
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.019
RMSD bond angle	1.996
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	PHASER
Refinement software	REFMAC (5.8.0425)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	19.950	19.950	1.300
High resolution limit [Å]	1.250	3.880	1.250
Rmerge	0.068	0.029	0.579
Rmeas	0.074	0.032	0.640
Total number of observations	447898
Number of reflections	63967	2334	6517
<I/σ(I)>	14.69	47.28	2.82
Completeness [%]	94.5

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.