8CNR
Hybrid Cluster Protein from the thermophilic methanogen Methanothermococcus thermolithotrophicus as isolated in a reduced state at 1.45-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 | 2020-02-25 |
Detector | DECTRIS PILATUS 2M-F |
Wavelength(s) | 1.23984 |
Spacegroup name | P 21 21 2 |
Unit cell lengths | 97.557, 102.454, 58.896 |
Unit cell angles | 90.00, 90.00, 90.00 |
Refinement procedure
Resolution | 51.060 - 1.450 |
R-factor | 0.1341 |
Rwork | 0.133 |
R-free | 0.15240 |
Structure solution method | SAD |
RMSD bond length | 0.009 |
RMSD bond angle | 1.288 |
Data reduction software | autoPROC |
Data scaling software | autoPROC |
Phasing software | SHELXDE |
Refinement software | PHENIX (1.20.1-4487) |
Data quality characteristics
Overall | Outer shell | |
Low resolution limit [Å] | 51.061 | 1.476 |
High resolution limit [Å] | 1.450 | 1.451 |
Rmerge | 0.119 | 1.193 |
Rmeas | 0.132 | 1.361 |
Rpim | 0.056 | 0.642 |
Number of reflections | 103697 | 4833 |
<I/σ(I)> | 9.5 | 1.4 |
Completeness [%] | 98.8 | 92.4 |
Redundancy | 5.3 | |
CC(1/2) | 0.996 | 0.510 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, SITTING DROP | 7.6 | 293.15 | Crystallisation was performed anaerobically at 20 degree Celsius using the sitting drop method on 96-Well MRC 2-Drop polystyrene Crystallisation Plates (SWISSCI) in a Coy tent containing a N2/H2 (97:3%) atmosphere. The reservoir chamber was filled with 90 ul of crystallisation condition and the drop was formed by spotting 0.55 ul protein with 0.55 ul of 20% (w/v) PEG 3,350 and 200 mM Magnesium formate. The protein was crystallized at 9.9 mg/ml in 25 mM Tris/HCl pH 7.6, 2 mM dithiothreitol and 10% (v/v) glycerol. Densities in the electron density map suggest a contamination from another crystallisation condition spatially close and containing 30% (v/v) 2-Methyl-2,4-pentanediol, 20 mM Calcium chloride and 100 mM Sodium acetate, pH 4.6. Crystals were cryo-protected by soaking in 20% (w/v) PEG 3,350 and 200mM Magnesium formate supplemented with 20% v/v glycerol for a few seconds. |