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Glutamine synthetase from Methermicoccus shengliensis at a resolution of 3.09 A

This is a non-PDB format compatible entry.

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SLS BEAMLINE X06DA
Synchrotron site	SLS
Beamline	X06DA
Temperature [K]	100
Detector technology	PIXEL
Collection date	2021-10-17
Detector	DECTRIS PILATUS 2M-F
Wavelength(s)	1.00002
Spacegroup name	P 43 3 2
Unit cell lengths	226.465, 226.465, 226.465
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	65.370 - 3.090
R-factor	0.1878
Rwork	0.186
R-free	0.21620
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.004
RMSD bond angle	0.678
Data reduction software	autoPROC
Data scaling software	STARANISO
Phasing software	PHASER
Refinement software	PHENIX ((1.20.1_4487: ???))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	130.750	3.172
High resolution limit [Å]	3.088	3.088
Rmerge	0.345	3.099
Rmeas	0.352	3.159
Rpim	0.068	0.607
Number of reflections	35711	1793
<I/σ(I)>	12.2	1.4
Completeness [%]	96.6	63.8
Redundancy	26.6	26.8
CC(1/2)	0.998	0.486

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.6	293.15	The protein was crystallized fresh without any freezing step, and obtained through the sitting drop method on a 96-Well MRC 2-Drop Crystallization Plates in polystyrene (SWISSCI, United Kingdom) under anaerobic conditions (N2:H2, gas ratio of 97:3). Crystallization was performed at 9 mg/ml glutamine synthetase in 25 mM Tris/HCl pH 7.6, 10% glycerol, and 2 mM dithiothreitol. The reservoir contained 90 ul precipitant (1.6 M sodium citrate tribasic dihydrate). 0.55 uL protein was mixed with 0.55 uL precipitant. Crystals were soaked in the precipitant supplemented with 30% v/v glycerol before freezing in liquid nitrogen.