3FYO

Crystal structure of the triple mutant (N23C/D247E/P249A) 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]	110
Detector technology	IMAGE PLATE
Collection date	2006-11-15
Detector	RIGAKU RAXIS IV++
Wavelength(s)	1.5418
Spacegroup name	P 21 21 21
Unit cell lengths	82.740, 86.197, 163.891
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	33.940 - 1.900
R-factor	0.218
Rwork	0.216
R-free	0.25700
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2qkf
RMSD bond length	0.010
RMSD bond angle	1.075
Data reduction software	d*TREK
Data scaling software	d*TREK
Phasing software	AMoRE
Refinement software	REFMAC

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	33.950	1.860
High resolution limit [Å]	1.800	1.800
Rmerge	0.065	0.472
Total number of observations		44603
Number of reflections	108582
<I/σ(I)>	9	2.3
Completeness [%]	99.5	100
Redundancy	4.15	4.09

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	5	297	21.0 mg/mL protein mixed 1:1 with reservoir liquor containing 100 mM NaOAc (pH 5.0), 2 mM PEP, 200 microM MnSO4 and 2.0 M NaCl. Immediately prior to data collection, crystal were harvested and soaked briefly in cryoprotectant solution, comprising 20% glycerol and 100 mM NaOAc (pH 5.0) and 2.0 M NaCl, VAPOR DIFFUSION, HANGING DROP, temperature 297K