8OON

Glutamine synthetase from Methanothermococcus thermolithotrophicus at a resolution of 2.43 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	2021-09-05
Detector	DECTRIS PILATUS 2M-F
Wavelength(s)	1.00004
Spacegroup name	C 2 2 21
Unit cell lengths	132.647, 230.244, 205.586
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	58.890 - 2.430
R-factor	0.233
Rwork	0.229
R-free	0.26820
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.004
RMSD bond angle	0.654
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]	76.675	2.567
High resolution limit [Å]	2.430	2.430
Rmerge	0.269	1.795
Rmeas	0.279	1.855
Rpim	0.073	0.464
Number of reflections	102291	5117
<I/σ(I)>	7.4	1.5
Completeness [%]	93.3	49.5
Redundancy	14.5	15.6
CC(1/2)	0.998	0.782

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.6	293.15	Protein was crystallized fresh without any freezing step by using 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 15 mg/ml 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 (20 % w/v polyethylene glycol 3,350 and 100 mM potassium sodium tartrate), the crystallization drop contained a mixture of 0.55 uL protein and 0.55 uL mother liquor. Crystals were soaked in the mother liquor supplemented with 25% v/v ethylene glycol prior to freezing in liquid nitrogen.