7NU2

Crystal Structure of Neisseria gonorrhoeae LeuRS E169G mutant in Complex with the Reaction Intermediate Leu-AMP

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	48.703, 82.410, 227.571
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	77.490 - 2.100
R-factor	0.1854
Rwork	0.183
R-free	0.23720
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]	77.490	77.490	2.180
High resolution limit [Å]	2.060	6.530	2.060
Rmerge	0.136	0.054	1.587
Rmeas	0.141	0.056	1.647
Rpim	0.038	0.016	0.435
Total number of observations	777663	25638	116426
Number of reflections	57491	2048	8269
<I/σ(I)>	12.4	37	1.8
Completeness [%]	100.0	100	100
Redundancy	13.5	12.5	14.1
CC(1/2)	0.998	0.998	0.737

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-leucine in an equilvalent precipitant solution supplemented with 22% (v/v) ethylene glycol.