4JTG

Crystal structure of F114R/R117A mutant of 3-deoxy-D-manno-octulosonate 8-phosphate synthase (KDO8PS) from Neisseria meningitidis

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	AUSTRALIAN SYNCHROTRON BEAMLINE MX1
Synchrotron site	Australian Synchrotron
Beamline	MX1
Temperature [K]	100
Detector technology	CCD
Collection date	2010-09-01
Detector	ADSC QUANTUM 210r
Wavelength(s)	0.95368
Spacegroup name	P 21 21 21
Unit cell lengths	81.590, 85.370, 163.350
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	33.576 - 1.850
R-factor	0.1624
Rwork	0.161
R-free	0.19090
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2qkf
RMSD bond length	0.020
RMSD bond angle	1.895
Data reduction software	MOSFLM
Data scaling software	SCALA (3.3.15)
Refinement software	REFMAC

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	33.576	33.576	1.950
High resolution limit [Å]	1.850	5.850	1.850
Rmerge		0.032	0.412
Total number of observations		21352	97126
Number of reflections	95642
<I/σ(I)>	20.7	16.5	1.9
Completeness [%]	97.9	98.6	96.8
Redundancy	7.4	6.5	7.1

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 1.6 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