1NX6

Crystal Structure of Aspartate Semialdehyde Dehydrogenase from Haemophilus influenzae as a Tetrahedral Hemithiocetal Reaction intermediate with Phosphate at 2.15 A

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 14-BM-C
Synchrotron site	APS
Beamline	14-BM-C
Temperature [K]	298
Detector technology	CCD
Collection date	2002-06-01
Detector	ADSC QUANTUM 4
Wavelength(s)	1.000
Spacegroup name	P 21 21 2
Unit cell lengths	113.748, 54.853, 57.691
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	40.500 - 2.150
R-factor	0.228
Rwork	0.228
R-free	0.28400
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1nwh
RMSD bond length	0.012
RMSD bond angle	23.800 *
Data reduction software	HKL-2000
Data scaling software	SCALEPACK
Phasing software	CNS
Refinement software	CNS (1.0)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	100.000	2.230
High resolution limit [Å]	2.150	2.150
Rmerge	0.068	0.353
Total number of observations	143627 *
Number of reflections	19724
<I/σ(I)>	27.8	5.6
Completeness [%]	99.9 *	100 *
Redundancy	7.06

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7 *	20 *	20-24% PEG 3350, 0.2 M Ammonium Acetate, 0.1 M Tris-HCl, pH 8.5, VAPOR DIFFUSION, HANGING DROP, temperature 293K

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	drop	protein	15 (mg/ml)
2	1	drop	HEPES	10 (mM)
3	1	drop	EDTA	1 (mM)
4	1	drop	dithiothreitol	1 (mM)	pH7.0
5	1	reservoir	PEG3350	24-28 (%)
6	1	reservoir	ammonium acetate	0.2 (M)
7	1	reservoir	Tris	0.1 (M)	pH8.5