4JTE

Crystal structure of F114A mutant 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	2007-03-05
Detector	RIGAKU RAXIS IV++
Wavelength(s)	1.5418
Spacegroup name	P 21 21 21
Unit cell lengths	81.355, 85.473, 163.057
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	39.950 - 1.900
R-factor	0.2305
Rwork	0.229
R-free	0.26690
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2qkf
RMSD bond length	0.017
RMSD bond angle	1.757
Data reduction software	d*TREK (9.4SSI)
Data scaling software	d*TREK (9.4SSI)
Refinement software	REFMAC

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	39.950	39.950	1.970
High resolution limit [Å]	1.900	4.090	1.900
Rmerge	0.065	0.030	0.601
Total number of observations		45354	33828
Number of reflections	88194
<I/σ(I)>	8.8	30.6	1.2
Completeness [%]	97.8	99.8	87
Redundancy	4.7	4.73	4.37

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