7NUB

Crystal Structure of Neisseria gonorrhoeae LeuRS L550G mutant in Complex with the Reaction Intermediate Leu-AMP

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SOLEIL BEAMLINE PROXIMA 1
Synchrotron site	SOLEIL
Beamline	PROXIMA 1
Temperature [K]	100
Detector technology	PIXEL
Collection date	2019-07-26
Detector	DECTRIS EIGER X 16M
Wavelength(s)	0.978565
Spacegroup name	P 21 21 21
Unit cell lengths	49.309, 81.748, 217.950
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	65.390 - 3.020
R-factor	0.224
Rwork	0.221
R-free	0.28710
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]	108.970	108.970	3.180
High resolution limit [Å]	3.020	9.540	3.020
Rmerge	0.236	0.062	1.520
Rmeas	0.247	0.065	1.581
Rpim	0.070	0.019	0.432
Total number of observations	220750	7648	34354
Number of reflections	17973	676	2582
<I/σ(I)>	12	29.8	2.7
Completeness [%]	98.9	99.9	100
Redundancy	12.3	11.3	13.3
CC(1/2)	0.996	0.999	0.765

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.