6Q8A

Neisseria gonorrhoeae Leucyl-tRNA Synthetase in Complex with 5'-O-(N-(L-Leucyl)-Sulfamoyl)Cytidine

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE ID23-1
Synchrotron site	ESRF
Beamline	ID23-1
Temperature [K]	100
Detector technology	PIXEL
Collection date	2018-10-03
Detector	DECTRIS PILATUS 6M
Wavelength(s)	0.976254
Spacegroup name	P 21 21 21
Unit cell lengths	49.151, 81.695, 225.429
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	112.710 - 2.110
R-factor	0.2019
Rwork	0.200
R-free	0.24730
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	A model generated from 3ZJU
Data reduction software	XDS
Data scaling software	Aimless (0.7.2)
Phasing software	PHASES
Refinement software	PHENIX (1.13_2998)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	112.710	112.710	2.200
High resolution limit [Å]	2.090	6.610	2.090
Rmerge	0.076	0.048	0.596
Rmeas	0.083	0.052	0.667
Rpim	0.033	0.021	0.290
Number of reflections	53482	1922	7011
<I/σ(I)>	13
Completeness [%]	97.5	98.5	89
Redundancy	5.8	5.7	4.8
CC(1/2)	0.998	0.997	0.724

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	8.5	293	Holo protein at 10 mg/mL in 10 mM Tris pH 7, 100 mM NaCl, 2.5 mM 2-mercaptoethanol was mixed with 0.1 M bis-tris propane pH 8.5, 0.1 M MgCl2, 20% w/v PEG 3350 and a crystal seed stock in a 0.75:1.0:0.25 (v/v) ratio. The seed stock was prepared in the same crystallization buffer. Suitable crystals were soaked with 2 mM 5'-O-(N-(L-Leucyl)-Sulfamoyl)Cytidine iin an equilvalent precipitant solution supplemented with 22% v/v ethylene glycol.