8RJA
Crystal structure of the F420-reducing formylmethanofuran dehydrogenase complex from the ethanotroph Candidatus Ethanoperedens thermophilum
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 | 2020-03-21 |
Detector | DECTRIS PILATUS 2M-F |
Wavelength(s) | 1.00000 |
Spacegroup name | P 1 21 1 |
Unit cell lengths | 107.627, 135.636, 149.902 |
Unit cell angles | 90.00, 90.49, 90.00 |
Refinement procedure
Resolution | 57.380 - 1.970 |
R-factor | 0.1774 |
Rwork | 0.176 |
R-free | 0.21000 |
Structure solution method | MOLECULAR REPLACEMENT |
RMSD bond length | 0.010 |
RMSD bond angle | 1.339 |
Data reduction software | autoPROC |
Data scaling software | autoPROC |
Phasing software | PHENIX |
Refinement software | PHENIX ((1.20.1_4487: ???)) |
Data quality characteristics
Overall | Outer shell | |
Low resolution limit [Å] | 57.380 | 2.187 |
High resolution limit [Å] | 1.970 | 1.973 |
Rmerge | 0.251 | 1.200 |
Rmeas | 0.271 | 1.296 |
Rpim | 0.101 | 0.486 |
Number of reflections | 167948 | 8397 |
<I/σ(I)> | 7.2 | 1.6 |
Completeness [%] | 93.0 | 64.3 |
Redundancy | 7 | 6.9 |
CC(1/2) | 0.989 | 0.621 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, SITTING DROP | 6.5 | 293.15 | Crystals were obtained by initial screening at 20 degrees Celsius using the sitting drop method on a 96-Well MRC 2-Drop Crystallization Plates in polystyrene (SWISSCI). The crystallization reservoir contained 90 ul of mother liquor, the crystallization drop contained a mixture of 0.6 ul protein at 4.64 mg.ml-1 and 0.6 ul precipitant. The protein was in 25 mM Tris/HCl buffer pH 7.6, 10 % (v/v) glycerol, 2 mM DTT, 1 mM FAD and 1 mM FMN. The crystal was obtained in a Coy tent under an N2/H2 atmosphere (96:4 %). The crystallization reservoir contained 200 mM sodium acetate trihydrate, 100 mM sodium cacodylate, pH 6.5, and 18 % (w/v) polyethylene glycol 8,000. The crystal was soaked in the crystallization solution supplemented with 30 % (v/v) glycerol before freezing in liquid nitrogen. |