1OZF

The crystal structure of Klebsiella pneumoniae acetolactate synthase with enzyme-bound cofactors

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 14-BM-D
Synchrotron site	APS
Beamline	14-BM-D
Temperature [K]	100
Detector technology	CCD
Collection date	2002-02-22
Detector	ADSC QUANTUM 4
Spacegroup name	C 2 2 21
Unit cell lengths	116.822, 160.573, 129.374
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	100.000 - 2.300
R-factor	0.165
Rwork	0.165
R-free	0.21500 *
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1jsc
RMSD bond length	0.005
RMSD bond angle	1.240 *
Data scaling software	SCALEPACK
Phasing software	AMoRE
Refinement software	CNS (1.0)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	100.000	2.380
High resolution limit [Å]	2.300	2.300
Rmerge	0.055 *	0.113 *
Total number of observations	195326 *
Number of reflections	48069	2370 *
<I/σ(I)>	21.5	5.6
Completeness [%]	87.5	43.7
Redundancy	4.1	2

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	Vapor diffusion, hanging drop *	7 *	17 *	PEG 8000, ethylene glycol, sodium HEPES, pH 7.6, VAPOR DIFFUSION, HANGING DROP, temperature 290K, pH 7.60

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	drop	enzyme	9 (mg/ml)
2	1	drop	potassium phosphate	50 (mM)	pH7.0
3	1	drop	ThDP	1 (mM)
4	1	drop		1 (mM)
5	1	drop	dithiothreitol	1 (mM)
6	1	reservoir	sodium HEPES	0.1 (M)	pH7.5-7.7
7	1	reservoir	PEG8000	6-8 (%(w/v))
8	1	reservoir	ethylene glycol	6-9 (%(v/v))