8OOQ

Glutamine synthetase from Methanothermococcus thermolithotrophicus in complex with 2-oxoglutarate and Mg at 2.91 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	2020-12-10
Detector	DECTRIS PILATUS 2M-F
Wavelength(s)	0.99999
Spacegroup name	P 1
Unit cell lengths	131.808, 131.927, 203.544
Unit cell angles	89.95, 89.86, 60.05

Refinement procedure

Resolution	43.740 - 2.910
R-factor	0.2553
Rwork	0.254
R-free	0.27800
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.010
RMSD bond angle	1.274
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]	203.543	3.124
High resolution limit [Å]	2.908	2.908
Rmerge	0.122	0.501
Rmeas	0.144	0.611
Rpim	0.076	0.344
Number of reflections	169224	8461
<I/σ(I)>	5.9	1.6
Completeness [%]	91.7	69.3
Redundancy	3.5	3
CC(1/2)	0.993	0.548

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	5	293.15	The protein was crystallized fresh without any freezing step, and crystallization was performed 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). The enzyme was crystallized at 3.4 mg/ml with a final concentration of 2 mM 2-oxoglutarate and 2 mM MgCl in 25 mM Tris/HCl pH 7.6, 10% v/v glycerol, 150 mM NaCl, and 2 mM dithiothreitol. The crystallization reservoir contained 90 ul of mother liquor (25 % w/v Polyethylene glycol 1,500 and 100 mM SPG (succinic acid, sodium dihydrogen phosphate, and glycine) buffer pH 5.0), and the crystallization drop contained 0.6 ul protein with ligands and 0.6 ul precipitant. Crystals were soaked in the mother liquor supplemented with 15 % v/v glycerol prior to freezing in liquid nitrogen.