7NTZ

Crystal Structure of Neisseria gonorrhoeae LeuRS L550A 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	2019-06-14
Detector	DECTRIS EIGER X 9M
Wavelength(s)	0.980118
Spacegroup name	P 21 21 21
Unit cell lengths	48.935, 81.446, 217.410
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	54.350 - 2.100
R-factor	0.2025
Rwork	0.202
R-free	0.22850
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]	108.710	108.710	2.160
High resolution limit [Å]	2.050	6.480	2.050
Rmerge	0.088	0.033	1.221
Rmeas	0.093	0.035	1.290
Rpim	0.030	0.011	0.412
Total number of observations	522825	17253	77713
Number of reflections	55849	1985	8013
<I/σ(I)>	14.1	46.7	1.7
Completeness [%]	100.0	99.8	100
Redundancy	9.4	8.7	9.7
CC(1/2)	0.999	0.999	0.701

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	8	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 8.0, 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.