5ODH
Heterodisulfide reductase / [NiFe]-hydrogenase complex from Methanothermococcus thermolithotrophicus soaked with heterodisulfide for 3.5 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 | 366.381, 97.104, 133.974 |
Unit cell angles | 90.00, 108.43, 90.00 |
Refinement procedure
Resolution | 48.552 - 2.200 |
R-factor | 0.2154 |
Rwork | 0.214 |
R-free | 0.24470 |
Structure solution method | MOLECULAR REPLACEMENT |
Starting model (for MR) | 5odc |
RMSD bond length | 0.006 |
RMSD bond angle | 1.527 |
Data reduction software | XDS |
Data scaling software | SCALA (3.3.22) |
Phasing software | PHASER |
Refinement software | PHENIX ((1.10.1_2155: ???)) |
Data quality characteristics
Overall | Outer shell | |
Low resolution limit [Å] | 48.552 | 2.320 |
High resolution limit [Å] | 2.200 | 2.200 |
Rmerge | 0.141 | 0.435 |
Rpim | 0.073 | 0.237 |
Number of reflections | 225651 | 32668 |
<I/σ(I)> | 8.4 | |
Completeness [%] | 99.8 | 99.5 |
Redundancy | 4.8 | 4.2 |
CC(1/2) | 0.992 | 0.451 |
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, 3% (v/v) DMSO, 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 crystals appeared after 1-2 weeks in this condition. The cryo-cool trapping experiment was performed as following: crystals were soaked for 3min30sec in the crystallization solution supplemented with 66 mM CoM-S-S-CoB. The crystal was cryo-protected by a soak in the crystallization solution supplemented by 30% glycerol and 10 mM CoM-S-S-CoB. |