2G8T

Indole-amidine Complexes with Bovine Trypsin

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 17-ID
Synchrotron site	APS
Beamline	17-ID
Temperature [K]	100
Detector technology	CCD
Collection date	2005-12-17
Detector	ADSC QUANTUM 4
Wavelength(s)	1
Spacegroup name	P 31 2 1
Unit cell lengths	54.525, 54.525, 107.807
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	43.250 - 1.410
R-factor	0.191
Rwork	0.190
R-free	0.20000
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.006
RMSD bond angle	1.400
Data reduction software	HKL-2000
Data scaling software	SCALEPACK
Phasing software	AMoRE
Refinement software	CNX (2002)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	43.250	1.500
High resolution limit [Å]	1.410	1.410
Number of reflections	36505
Completeness [%]	99.8	99.4

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	6.5	293	Trypsin, final concentration of 0.6 mM, was dissolved in 20 mM HEPES pH 7.0, 10 mM calcium chloride, 3% DMSO, and 3 mM of ligand of interest (compounds 3, 6, and 9). The well solution consisted of 100 mM Cacodylate pH 6.5, 200 mM ammonium sulfate, 50% PEG (17% to 28%). The ligands were co crystallized with protein using vapor diffusion and hanging drop method. The drops were 1:1 (protein:well solution) and the plates were stored at 20 degrees centigrade. The crystals were generally present within two days. The cryo-protectant consisted of adding 75 l of an adjusted well solution, adjusted with PEG 8000 to 0.5% higher, and 25 l of glycerol. drop 1:1, VAPOR DIFFUSION, HANGING DROP, temperature 293K