6NCS

Crystal structure of N-acetylneuraminic acid (Sialic acid) synthetase from Leptospira borgpetersenii serovar Hardjo-bovis in complex with citrate

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	RIGAKU FR-E+ SUPERBRIGHT
Temperature [K]	100
Detector technology	CCD
Collection date	2017-11-10
Detector	RIGAKU SATURN 944+
Wavelength(s)	1.5418
Spacegroup name	P 1 21 1
Unit cell lengths	69.880, 49.120, 88.850
Unit cell angles	90.00, 112.41, 90.00

Refinement procedure

Resolution	43.375 - 1.800
R-factor	0.1573
Rwork	0.156
R-free	0.19830
Structure solution method	SAD
RMSD bond length	0.006
RMSD bond angle	0.824
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	PHASER
Refinement software	PHENIX (dev_3339)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	43.375	43.375	1.850
High resolution limit [Å]	1.800	8.050	1.800
Rmerge	0.057	0.037	0.398
Rmeas	0.060	0.039	0.428
Total number of observations	517998
Number of reflections	50583	632	3381
<I/σ(I)>	25	62.58	4.23
Completeness [%]	97.1	98.8	88.2
Redundancy	10.241	12.704	6.728
CC(1/2)	0.999	0.999	0.941

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.5	285	Molecular Dimensions Morpheus screen, condition G8: 12.5% w/v PEG 1000, 12.5% w/v PEG 3350, 12.5% v/v MPD: 20mM each: sodium formate, ammonium acetate, trisodium citrate, sodium potassium L-tartrate, sodium oxamate: 100mM MOPS/HEPES-Na pH 7.5: LpboA.19655.b.B1.PS38488 at 22.9mg/ml. For phasing, the crystal was incubated for 20sec in reservoir solution with additional 10% of 5M NaI in ethylene glycol (500mM final concentration). Cryo: direct: tray: 303852g8: puck hyz8-5.