7NU7

Crystal Structure of Neisseria gonorrhoeae LeuRS in Complex with ATP in Conformation 2

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE MASSIF-3
Synchrotron site	ESRF
Beamline	MASSIF-3
Temperature [K]	100
Detector technology	PIXEL
Collection date	2018-12-02
Detector	DECTRIS EIGER X 4M
Wavelength(s)	0.967700
Spacegroup name	P 21 21 21
Unit cell lengths	49.141, 82.728, 228.468
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	48.040 - 2.310
R-factor	0.2082
Rwork	0.205
R-free	0.26400
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	6q89
Data reduction software	XDS
Data scaling software	Aimless (0.7.2)
Phasing software	PHASER
Refinement software	PHENIX (1.17.1_3660)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	82.730	82.730	2.360
High resolution limit [Å]	2.240	7.090	2.240
Rmerge	0.092	0.037	1.046
Rmeas	0.099	0.040	1.120
Rpim	0.036	0.015	0.394
Total number of observations	337512	10858	51558
Number of reflections	45696	1626	6549
<I/σ(I)>	13.6	39.3	1.7
Completeness [%]	99.9	99.8	100
Redundancy	7.4	6.7	7.9
CC(1/2)	0.998	0.999	0.656

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 incubated with 10 mM ATP on ice for half an hour. The premixture was further 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 cryo-protected in an equilvalent precipitant solution supplemented with 22% v/v ethylene glycol.