7NUC

Crystal Structure of Neisseria gonorrhoeae LeuRS L550G mutant in Complex with ATP and L-leucinol

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	48.912, 81.588, 216.971
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	76.367 - 2.770
R-factor	0.189
Rwork	0.185
R-free	0.26440
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 (1.15.2_3472)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	108.490	108.490	2.900
High resolution limit [Å]	2.750	8.710	2.750
Rmerge	0.368	0.122	1.576
Rmeas	0.391	0.130	1.670
Rpim	0.131	0.045	0.549
Total number of observations	203391	6721	30555
Number of reflections	23377	862	3349
<I/σ(I)>	6.4	14.2	2
Completeness [%]	100.0	99.9	100
Redundancy	8.7	7.8	9.1
CC(1/2)	0.979	0.987	0.715

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.