6HR8
HMG-CoA reductase from Methanothermococcus thermolithotrophicus in complex with NADPH at 2.9 A resolution
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | SYNCHROTRON |
Source details | ESRF BEAMLINE ID23-1 |
Synchrotron site | ESRF |
Beamline | ID23-1 |
Temperature [K] | 100 |
Detector technology | PIXEL |
Collection date | 2017-02-12 |
Detector | DECTRIS PILATUS 6M |
Wavelength(s) | 0.97662 |
Spacegroup name | P 4 21 2 |
Unit cell lengths | 231.790, 231.790, 98.714 |
Unit cell angles | 90.00, 90.00, 90.00 |
Refinement procedure
Resolution | 24.987 - 2.900 |
R-factor | 0.2127 |
Rwork | 0.211 |
R-free | 0.24260 |
Structure solution method | MOLECULAR REPLACEMENT |
Starting model (for MR) | 6hr7 |
RMSD bond length | 0.003 |
RMSD bond angle | 0.653 |
Data reduction software | XDS (3.3.22) |
Data scaling software | SCALA |
Phasing software | PHASER (2.6.0) |
Refinement software | PHENIX ((1.10.1_2155: ???)) |
Data quality characteristics
Overall | Outer shell | |
Low resolution limit [Å] | 25.000 | 3.060 |
High resolution limit [Å] | 2.900 | 2.900 |
Rmerge | 0.095 | 1.285 |
Rmeas | 0.102 | 1.377 |
Rpim | 0.037 | 0.491 |
Number of reflections | 59867 | 8592 |
<I/σ(I)> | 13.3 | 1.6 |
Completeness [%] | 99.8 | 100 |
Redundancy | 7.3 | 7.7 |
CC(1/2) | 0.999 | 0.427 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, SITTING DROP | 8.5 | 291 | HMGR at 10 mg/ml was supplemented with a final concentration of 2 mM NADPH. The sample was centrifuged for 5 minute at 13.000 RPM to remove any aggregates and dust. 0.7 ul of protein sample was spotted on a 96-well 2-drop MRC Crystallization Plates (Molecular Dimensions, Suffolk, UK) and 0.7 ul of reservoir solution was mixed. The best crystals were obtained after several month using a solution containing 40% v/v PEG 400, 100 mM Tris PH 8.5 and 200 mM Li2SO4. |