1RF4

Structural Studies of Streptococcus pneumoniae EPSP Synthase, Tetrahedral intermediate Bound State

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SSRL BEAMLINE BL9-2
Synchrotron site	SSRL
Beamline	BL9-2
Temperature [K]	100
Detector technology	CCD
Collection date	2001-12-05
Detector	ADSC QUANTUM 315
Wavelength(s)	0.98
Spacegroup name	P 1 21 1
Unit cell lengths	82.495, 149.705, 88.011
Unit cell angles	90.00, 91.22, 90.00

Refinement procedure

Resolution	10.000 - 2.200
R-factor	0.214
Rwork	0.196
R-free	0.23600
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.021
RMSD bond angle	1.810
Data scaling software	SCALA
Phasing software	AMoRE
Refinement software	X-PLOR (3.851)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	30.000	2.300
High resolution limit [Å]	2.200	2.200
Rmerge	0.041 *	0.180 *
Total number of observations	822394 *
Number of reflections	103059
Completeness [%]	97.2	91

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7.5 *	4 *	PEG, EG, NaCl, Na/K phosphate, pH 6.2, VAPOR DIFFUSION, HANGING DROP, temperature 277K

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	reservoir	PEG8000	10 (%)
2	1	reservoir	EG	8 (%)
3	1	reservoir		0.2 (M)
4	1	reservoir	sodium pootassium phosphate	0.1 (M)	pH6.2
5	1	reservoir	ligand	5 (mM)
6	1	drop	protein	60 (mg/ml)
7	1	drop	Tris-HCl	10 (mM)	pH7.5
8	1	drop		100 (mM)