8OOZ

Glutamine synthetase from Methermicoccus shengliensis in complex with MgATP at 2.7 A resolution

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	2021-11-27
Detector	DECTRIS PILATUS 2M-F
Wavelength(s)	0.97949
Spacegroup name	P 1 21 1
Unit cell lengths	131.548, 197.399, 135.171
Unit cell angles	90.00, 94.89, 90.00

Refinement procedure

Resolution	49.100 - 2.700
R-factor	0.1972
Rwork	0.196
R-free	0.21720
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.011
RMSD bond angle	1.348
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]	78.845	2.878
High resolution limit [Å]	2.700	2.700
Rmerge	0.062	0.688
Rmeas	0.070	0.784
Rpim	0.033	0.370
Number of reflections	128572	6429
<I/σ(I)>	17.4	2
Completeness [%]	88.1	89.8
Redundancy	4.3	4.4
CC(1/2)	0.999	0.701

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% v/v 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 for 4 min in the crystallization solution supplemented with 10 mM ATP/Mg2+/sodium glutamate and then back soaked in the crystallization solution supplemented with 25% glycerol prior to freezing in liquid nitrogen.