8OOO
Glutamine synthetase from Methanothermococcus thermolithotrophicus in complex with 2-oxoglutarate and MgATP at 2.15 A resolution
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 | 2020-08-27 |
Detector | DECTRIS PILATUS 2M-F |
Wavelength(s) | 1.30511 |
Spacegroup name | P 1 |
Unit cell lengths | 112.337, 131.773, 131.507 |
Unit cell angles | 60.04, 87.72, 67.34 |
Refinement procedure
Resolution | 42.880 - 2.149 |
R-factor | 0.1832 |
Rwork | 0.182 |
R-free | 0.21400 |
Structure solution method | MOLECULAR REPLACEMENT |
RMSD bond length | 0.009 |
RMSD bond angle | 0.930 |
Data reduction software | autoPROC |
Data scaling software | STARANISO |
Phasing software | PHASER |
Refinement software | BUSTER (2.10.4 (21-NOV-2022)) |
Data quality characteristics
Overall | Outer shell | |
Low resolution limit [Å] | 110.034 | 2.297 |
High resolution limit [Å] | 2.148 | 2.148 |
Rmerge | 0.110 | 0.950 |
Rmeas | 0.130 | 1.122 |
Rpim | 0.068 | 0.590 |
Number of reflections | 248419 | 12420 |
<I/σ(I)> | 9.3 | 1.3 |
Completeness [%] | 92.2 | 58.8 |
Redundancy | 3.6 | 3.4 |
CC(1/2) | 0.995 | 0.569 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, SITTING DROP | 7.6 | 293.15 | The enzyme was crystallized at 3.7 mg/ml with a final concentration of 2 mM sodium 2-oxoglutarate, 2 mM ATP and 2 mM MgCl in 25 mM Tris/HCl pH 7.6, 10% v/v glycerol, 150 mM NaCl, 2 mM dithiothreitol. 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 crystallization reservoir contained 90 ul of mother liquor (20 % w/v polyethylene glycol 3,350 and 200 mM sodium fluoride). The crystallization drop contained 0.6 ul protein with ligands and 0.6 ul precipitant. Crystals were soaked in the mother liquor supplemented with 20 % v/v glycerol prior to freezing in liquid nitrogen. |