1B4E

X-ray structure of 5-aminolevulinic acid dehydratase complexed with the inhibitor levulinic acid

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SRS BEAMLINE PX9.6
Synchrotron site	SRS
Beamline	PX9.6
Temperature [K]	100
Detector technology	IMAGE PLATE
Collection date	1994-03-15
Detector	MAR scanner 300 mm plate
Spacegroup name	I 4 2 2
Unit cell lengths	126.700, 126.700, 141.600
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	7.700 * - 2.000
Rwork	0.188
R-free	0.25500
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	YEAST ALAD
RMSD bond length	0.016
RMSD bond angle	0.032
Data reduction software	MOSFLM
Data scaling software	CCP4 ((AGROVATA)
Phasing software	CCP4 ((ALMN)
Refinement software	RESTRAIN

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	7.700 *	2.100
High resolution limit [Å]	2.000	2.000
Rmerge	0.063	0.297
Number of reflections	33275
<I/σ(I)>	8.8	2.1
Completeness [%]	85.8	70.8
Redundancy	2.5	1.6

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	8.2		7MG/ML ENZYME AND 2 - 5 % SATURATED AMMONIUM SULPHATE AS PRECIPITANT. 0.2 M TRIS-HCL USED TO BUFFER THE PH BETWEEN 8.1 AND 8.4. 15MM LEVULINIC ACID, 40UM ZINC SULPHATE AND 4MM BETA- MERCAPTOETHANOL WERE PRESENT., pH 8.2, VAPOR DIFFUSION, HANGING DROP

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	drop	enzyme	7 (mg/ml)
2	1	drop	Tris-HCl	0.2 (M)
3	1	drop	levulinic acid	15 (mM)
4	1	drop	zinc sulfate	0.04 (M)
5	1	drop	mercaptoethanol	4 (mM)
6	1	reservoir	ammonium sulfate	2-5 (%sat)