1KNK

Crystal Structure of 2-C-methyl-D-erythritol 2,4-cyclodiphosphate Synthase (ispF) from E. coli involved in Mevalonate-Independent Isoprenoid Biosynthesis

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE ID14-4
Synchrotron site	ESRF
Beamline	ID14-4
Temperature [K]	100
Detector technology	CCD
Collection date	2001-10-10
Detector	ADSC QUANTUM 4
Wavelength(s)	0.9793
Spacegroup name	I 21 3
Unit cell lengths	145.630, 145.630, 145.630
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	24.740 - 2.800
Rwork	0.232
R-free	0.25300 *
Structure solution method	Rigid body
Starting model (for MR)	1knj
RMSD bond length	0.010
RMSD bond angle	24.400 *
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	CNS
Refinement software	CNS

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	99.000 *	2.900
High resolution limit [Å]	2.800	2.800
Rmerge	0.091 *	0.415 *
Total number of observations	41619 *
Number of reflections	11929 *
<I/σ(I)>	15.7	5
Completeness [%]	93.6	96.9
Redundancy	3.55 *	3.3

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	6.5	4 *	(NH4)2SO4, PEG 400, NaI, PIPES, pH 6.5, VAPOR DIFFUSION, HANGING DROP, temperature 294K

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	drop	protein	13.5 (mg/ml)
2	1	reservoir	ammonium sulfate	0.8 (M)
3	1	reservoir	PEG400	1 (%(w/v))
4	1	reservoir	sodium iodide	0.2 (M)
5	1	reservoir	DL-dithiothreitol	2 (mM)
6	1	reservoir	PIPES	0.1 (M)	pH6.5