5ODI
Heterodisulfide reductase / [NiFe]-hydrogenase complex from Methanothermococcus thermolithotrophicus cocrystallized with CoM-SH
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) | 0.97916 |
Spacegroup name | C 1 2 1 |
Unit cell lengths | 370.528, 97.077, 135.149 |
Unit cell angles | 90.00, 109.43, 90.00 |
Refinement procedure
Resolution | 48.540 - 2.400 |
R-factor | 0.2035 |
Rwork | 0.202 |
R-free | 0.24080 |
Structure solution method | MOLECULAR REPLACEMENT |
Starting model (for MR) | 5odc |
RMSD bond length | 0.006 |
RMSD bond angle | 1.050 |
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] | 48.540 | 2.530 |
High resolution limit [Å] | 2.400 | 2.400 |
Rmerge | 0.189 | 0.507 |
Rpim | 0.071 | 0.188 |
Number of reflections | 176807 | 25737 |
<I/σ(I)> | 7.2 | 3.6 |
Completeness [%] | 100.0 | 100 |
Redundancy | 8 | 8.2 |
CC(1/2) | 0.992 | 0.332 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, SITTING DROP | 8.5 | 294 | 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, 1% (w/v) polyethylene glycol 3350, 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, 10 mM CoM-SH and 1 ul of precipitant. The crystals appear after 1-2 weeks and are cryoprotected in the crystallization solution supplemented with 30% glycerol |