7H63

THE 1.65 A CRYSTAL STRUCTURE OF HUMAN CHYMASE IN COMPLEX WITH 4-[(5-fluoro-3-propan-2-yl-1H-indol-2-yl)-phenylmethyl]-3-hydroxy-2-propan-2-yl-1,2-dihydropyrrol-5-one (VINYLOGOUS ACID)

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	2005-11-25
Detector	MAR scanner 345 mm plate
Wavelength(s)	1.5418
Spacegroup name	P 43
Unit cell lengths	74.810, 74.810, 49.520
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	19.910 - 1.650
R-factor	0.17314
Rwork	0.172
R-free	0.19317
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.004
RMSD bond angle	1.174
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]	19.910	19.910	1.770
High resolution limit [Å]	1.650	4.860	1.650
Rmerge	0.062	0.033	0.540
Rmeas	0.066	0.035	0.588
Total number of observations	231352
Number of reflections	33005	1340	6173
<I/σ(I)>	22.18	52.33	3.5
Completeness [%]	99.6

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.