7NU1

Crystal Structure of Neisseria gonorrhoeae LeuRS E169G mutant

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	2020-07-24
Detector	DECTRIS EIGER X 9M
Wavelength(s)	0.980114
Spacegroup name	P 21 21 21
Unit cell lengths	49.049, 81.876, 226.159
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	19.940 - 2.510
R-factor	0.2064
Rwork	0.203
R-free	0.26170
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]	19.940	19.940	2.650
High resolution limit [Å]	2.510	7.950	2.510
Rmerge	0.153	0.056	1.041
Rmeas	0.164	0.061	1.109
Rpim	0.057	0.022	0.378
Total number of observations	258540	7639	38354
Number of reflections	31960	1061	4573
<I/σ(I)>	9.5	25	2.2
Completeness [%]	99.8	93.6	100
Redundancy	8.1	7.2	8.4
CC(1/2)	0.994	0.993	0.756

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 cryoprotected in an equilvalent precipitant solution supplemented with 22% v/v ethylene glycol.