1MX5

Crystal Structure of Human Liver Carboxylesterase in complexed with homatropine, a cocaine analogue

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SSRL BEAMLINE BL7-1
Synchrotron site	SSRL
Beamline	BL7-1
Temperature [K]	200
Detector technology	IMAGE PLATE
Collection date	2002-03-31
Detector	MARRESEARCH
Wavelength(s)	1.5418
Spacegroup name	P 1 21 1
Unit cell lengths	55.400, 178.800, 199.600
Unit cell angles	90.00, 90.20, 90.00

Refinement procedure

Resolution	20.000 * - 2.800
R-factor	0.158
Rwork	0.158
R-free	0.22200 *
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.006
RMSD bond angle	22.800 *
Data reduction software	MOSFLM
Data scaling software	SCALA
Phasing software	AMoRE
Refinement software	CNS (1.0)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	20.000
High resolution limit [Å]	2.800	2.800 *
Rmerge	0.050 *	0.114 *
Total number of observations	292059 *
Number of reflections	82155
Completeness [%]	92.5	80.6 *
Redundancy	3.6 *

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.4 *	22 *	PEG3350, Glycerol, lithium sulfate, lithium chloride, sodium chloride, sodium citrate, pH 5.5, VAPOR DIFFUSION, SITTING DROP, temperature 298K

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	drop	protein	3 (mg/ml)
2	1	drop	HEPES	50 (mM)	pH7.4
3	1	drop	naloxone methiodide	10 (mM)	or 100mM homatropine
4	1	reservoir	PEG3350	10 (%(w/v))
5	1	reservoir		0.4 (M)
6	1	reservoir		0.1 (M)
7	1	reservoir		0.1 (M)
8	1	reservoir	citrate	0.1 (M)	pH5.5
9	1	reservoir	glycerol	5 (%)