5ODR
Heterodisulfide reductase / [NiFe]-hydrogenase complex from Methanothermococcus thermolithotrophicus soaked with heterodisulfide for 2 minutes.
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 | 366.219, 96.920, 134.450 |
Unit cell angles | 90.00, 108.30, 90.00 |
Refinement procedure
Resolution | 90.260 - 2.200 |
R-factor | 0.2048 |
Rwork | 0.204 |
R-free | 0.22480 |
Structure solution method | MOLECULAR REPLACEMENT |
Starting model (for MR) | 5odc |
RMSD bond length | 0.006 |
RMSD bond angle | 0.900 |
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] | 93.360 | 2.320 |
High resolution limit [Å] | 2.200 | 2.200 |
Rmerge | 0.181 | 0.897 |
Rpim | 0.096 | 0.496 |
Number of reflections | 224984 | 32472 |
<I/σ(I)> | 4.6 | 1.3 |
Completeness [%] | 99.4 | 98.7 |
Redundancy | 4.3 | 4 |
CC(1/2) | 0.983 | 0.563 |
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. Crystal was soaked for 2 minutes in the crystallization solution supplemented with 66 mM of heterodisulfide (CoM-S-S-CoB). Then, the crystal was transferred in a solution containing the mother liquor with 30% glycerol and 10 mM CoM-S-S-CoB prior freezing. |