1KNJ

Co-Crystal Structure of 2-C-methyl-D-erythritol 2,4-cyclodiphosphate Synthase (ispF) from E. coli Involved in Mevalonate-Independent Isoprenoid Biosynthesis, Complexed with CMP/MECDP/Mn2+

Experimental procedure

Experimental method	MAD
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE BM30A
Synchrotron site	ESRF
Beamline	BM30A
Temperature [K]	100
Detector technology	CCD
Collection date	2001-03-26
Detector	MARRESEARCH
Wavelength(s)	0.9795, 0.9800, 0.9724
Spacegroup name	I 21 3
Unit cell lengths	144.254, 144.254, 144.254
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	14.710 - 2.800
Rwork	0.241
R-free	0.25800 *
Structure solution method	MAD
RMSD bond length	0.010
RMSD bond angle	23.600 *
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	2.870
High resolution limit [Å]	2.800 *	2.800
Rmerge	0.066 *	0.660
Total number of observations	96785 *
Number of reflections	24118 *
<I/σ(I)>	19	482
Completeness [%]	99.3	100
Redundancy	4.1	4.1

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