3STE

Crystal structure of a mutant (Q202A) of 3-deoxy-D-manno-octulosonate 8-phosphate synthase (KDO8PS) from Neisseria meningitidis

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	RIGAKU MICROMAX-007
Temperature [K]	120
Detector technology	IMAGE PLATE
Collection date	2009-12-14
Detector	RIGAKU RAXIS IV++
Wavelength(s)	1.5418
Spacegroup name	P 21 21 21
Unit cell lengths	82.002, 85.709, 163.406
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	30.600 - 2.050
R-factor	0.207
Rwork	0.205
R-free	0.24740
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2qkf
RMSD bond length	0.010
RMSD bond angle	1.102
Data reduction software	d*TREK
Data scaling software	d*TREK
Refinement software	REFMAC

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	30.600	30.560	2.120
High resolution limit [Å]	2.050	4.410	2.050
Rmerge		0.037	0.383
Number of reflections	72315
<I/σ(I)>		23.2	2.7
Completeness [%]	99.3	96.5	99.9
Redundancy	4.22	4.04	4.17

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	4.6	297	20 mg/mL protein (in 10 mM BTP pH 7.5) mixed 1:1 with reservoir liquor containing 100 mM NaOAc (pH 4.6) and 0.6-3.0 M NaCl. Immediately prior to data collection, crystals were harvested and soaked briefly in cryoprotectant solution, comprising 20% glycerol and the reservoir solution, Vapor diffusion, hanging drop, temperature 297K