1XG3

Crystal structure of the C123S 2-methylisocitrate lyase mutant from Escherichia coli in complex with the reaction product, Mg(II)-pyruvate and succinate

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	RIGAKU RU300
Temperature [K]	100
Detector technology	IMAGE PLATE
Collection date	2004-01-18
Detector	RIGAKU RAXIS IV
Wavelength(s)	1.5418
Spacegroup name	C 1 2 1
Unit cell lengths	159.358, 85.993, 99.597
Unit cell angles	90.00, 108.28, 90.00

Refinement procedure

Resolution	25.850 - 1.900
Rwork	0.161
R-free	0.19500
Structure solution method	FOURIER SYNTHESIS
Starting model (for MR)	structure of the C123S mutant enzyme in complex with isocitrate
RMSD bond length	0.007
RMSD bond angle	1.300
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	CNS
Refinement software	CNS

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	25.850	1.970
High resolution limit [Å]	1.900	1.900
Rmerge	0.025	0.084
Number of reflections	89039
<I/σ(I)>	20.3
Completeness [%]	88.5	67.5
Redundancy	2.82

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	6	298	30% PEG3350, 0.1M (NH)2SO4, 0.1M MES (pH6.0). 5mM Mg(II), 5mM isocitrate, followed by overnight soaking of a crystal in 35% PEG3350, 100 mM succinate, 100 mM pyruvate, 5mM Mg(II), VAPOR DIFFUSION, HANGING DROP, temperature 298.0K