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.