7NU3

Crystal Structure of Neisseria gonorrhoeae LeuRS E169G mutant in Complex with ATP and L-leucinol

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SOLEIL BEAMLINE PROXIMA 2
Synchrotron site	SOLEIL
Beamline	PROXIMA 2
Temperature [K]	100
Detector technology	PIXEL
Collection date	2020-02-14
Detector	DECTRIS EIGER X 9M
Wavelength(s)	0.918398
Spacegroup name	P 21 21 21
Unit cell lengths	48.878, 82.214, 227.207
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	66.600 - 2.270
R-factor	0.2018
Rwork	0.200
R-free	0.24870
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	6q89
Data reduction software	XDS
Data scaling software	Aimless (0.7.4)
Phasing software	PHASER
Refinement software	PHENIX (1.17.1_3660)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	66.600	66.600	2.350
High resolution limit [Å]	2.230	7.060	2.230
Rmerge	0.151	0.049	1.582
Rmeas	0.157	0.051	1.645
Rpim	0.043	0.015	0.448
Total number of observations	604905	19275	86903
Number of reflections	45572	1621	6532
<I/σ(I)>	12.4	36.2	1.9
Completeness [%]	100.0	99.9	100
Redundancy	13.3	11.9	13.3
CC(1/2)	0.998	0.998	0.680

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 5 mM ATP and 5 mM L-leucinol in an equilvalent precipitant solution supplemented with 22% (v/v) ethylene glycol.