6ESQ
Structure of the acetoacetyl-CoA thiolase/HMG-CoA synthase complex from Methanothermococcus thermolithotrophicus soaked with acetyl-CoA
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 | 2016-06-27 |
Detector | DECTRIS PILATUS 6M |
Wavelength(s) | 1.64861 |
Spacegroup name | P 21 21 21 |
Unit cell lengths | 107.603, 145.176, 230.891 |
Unit cell angles | 90.00, 90.00, 90.00 |
Refinement procedure
Resolution | 49.290 - 2.950 |
R-factor | 0.184 |
Rwork | 0.182 |
R-free | 0.23000 |
Structure solution method | MOLECULAR REPLACEMENT |
RMSD bond length | 0.005 |
RMSD bond angle | 0.580 |
Data reduction software | XDS |
Data scaling software | SCALA |
Phasing software | PHASER |
Refinement software | BUSTER (2.10.3) |
Data quality characteristics
Overall | Outer shell | |
Low resolution limit [Å] | 49.300 | 3.110 |
High resolution limit [Å] | 2.950 | 2.950 |
Rmerge | 0.143 | 2.024 |
Rpim | 0.040 | 0.577 |
Number of reflections | 76363 | 10942 |
<I/σ(I)> | 14.9 | 1.3 |
Completeness [%] | 99.4 | 98.8 |
Redundancy | 13.3 | 13.1 |
CC(1/2) | 0.999 | 0.481 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, SITTING DROP | 8.5 | 293 | Crystallization was done under air using the sitting drop method (in a 24-well junior clover plate from Jena Bioscience). The crystallization reservoir contained 100 mM Tris/HCl pH 8.0, 25-28% v/v pentaerythritol ethoxylate (15/4 EO/OH, average Mn about 797) and 50 mM MgCl2. Crystallization drop contained 1 to 2 ul of the purified fraction containing Thiolase/HMGCS complex at 50-60 mg/ml (pure at 60%) and 1 ul of precipitant. The crystal was soaked in the crystallization condition supplemented 100 mM acetyl-CoA for 1 min 30 sec. |