5ODQ
Heterodisulfide reductase / [NiFe]-hydrogenase complex from Methanothermococcus thermolithotrophicus soaked with bromoethanesulfonate.
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | SYNCHROTRON |
Source details | SLS BEAMLINE X10SA |
Synchrotron site | SLS |
Beamline | X10SA |
Temperature [K] | 100 |
Detector technology | PIXEL |
Collection date | 2016-06-23 |
Detector | DECTRIS PILATUS 6M |
Wavelength(s) | 0.91941 |
Spacegroup name | C 1 2 1 |
Unit cell lengths | 366.325, 97.352, 133.276 |
Unit cell angles | 90.00, 108.37, 90.00 |
Refinement procedure
Resolution | 48.680 - 2.150 |
R-factor | 0.1973 |
Rwork | 0.196 |
R-free | 0.21860 |
Structure solution method | MOLECULAR REPLACEMENT |
Starting model (for MR) | 5odc |
RMSD bond length | 0.006 |
RMSD bond angle | 0.940 |
Data reduction software | XDS |
Data scaling software | SCALA (3.3.22) |
Phasing software | PHASER |
Refinement software | BUSTER (2.10.1) |
Data quality characteristics
Overall | Outer shell | |
Low resolution limit [Å] | 48.850 | 2.270 |
High resolution limit [Å] | 2.150 | 2.150 |
Rmerge | 0.210 | 0.606 |
Rpim | 0.086 | 0.249 |
Number of reflections | 241487 | 35106 |
<I/σ(I)> | 8 | 3.3 |
Completeness [%] | 100.0 | 100 |
Redundancy | 6.9 | 6.8 |
CC(1/2) | 0.991 | 0.327 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, SITTING DROP | 8.5 | 294 | All crystallization was performed in an anaerobic chamber (95% N2/5% H2) with anoxic solution using the sitting drop method (96-well 2-drop MRC Crystallization Plates in polystyrene, Molecular Dimensions, Suffolk, UK). The crystallization reservoir contained 100 mM Tris/HCl, pH 8.5, 30% (w/v) polyethylene glycol 4000, and 200 mM Na acetate trihydrate. Crystallization drop contained 1 ul HdrABC-MvhAGD at 25 mg/ml premixed with 2 mM FAD and 1 ul of precipitant. The soaking experiment with the bromoethanesulfonate was done by soaking the crystals in the same crystallization solution supplemented with 66 mM of bromoethanesulfonate for 7 min. The crystal has been cryo-protected using the the mother liquor solution supplemented with 30% glycerol and 10 mM bromoethanesulfonate. |