6ET9

Structure of the acetoacetyl-CoA-thiolase/HMG-CoA-synthase complex from Methanothermococcus thermolithotrophicus at 2.75 A

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE ID29
Synchrotron site	ESRF
Beamline	ID29
Temperature [K]	100
Detector technology	PIXEL
Collection date	2016-02-27
Detector	DECTRIS PILATUS 6M-F
Wavelength(s)	1.38546
Spacegroup name	P 21 21 21
Unit cell lengths	106.617, 144.014, 230.585
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	48.387 - 2.750
R-factor	0.1919
Rwork	0.190
R-free	0.22470
RMSD bond length	0.008
RMSD bond angle	0.825
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]	48.900	2.900
High resolution limit [Å]	2.750	2.750
Rmerge	0.279	1.249
Rpim	0.079	0.354
Number of reflections	92965	13426
<I/σ(I)>	8.6	2.1
Completeness [%]	100.0	100
Redundancy	13.4	13.3
CC(1/2)	0.993	0.324

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 Molecular weight, about 797 Da) 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%) with 10 mM Tb-Xo4 mixed with 1 ul of precipitant.