9G7I

Structure of carbon monoxide dehydrogenase/acetyl-CoA synthase (CODH/ACS) in complex with acetyl-Coenyzme A from Clostridium autoethanogenum

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SLS BEAMLINE X06DA
Synchrotron site	SLS
Beamline	X06DA
Temperature [K]	100
Detector technology	PIXEL
Collection date	2021-01-29
Detector	DECTRIS PILATUS 2M-F
Wavelength(s)	1.00003
Spacegroup name	P 42 21 2
Unit cell lengths	298.077, 298.077, 127.526
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	49.680 - 2.930
R-factor	0.1917
Rwork	0.190
R-free	0.22020
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.010
RMSD bond angle	1.367
Data reduction software	autoPROC
Data scaling software	autoPROC
Phasing software	PHASER
Refinement software	PHENIX ((1.20.1_4487: ???))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	59.610	3.160
High resolution limit [Å]	2.930	2.930
Rmerge	0.207	2.423
Rmeas	0.211	2.477
Rpim	0.045	0.513
Number of reflections	99666	4983
<I/σ(I)>	14.5	1.6
Completeness [%]	96.1	73
Redundancy	22.1	23.3
CC(1/2)	0.998	0.593

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	8.5	293.15	Crystallization was performed anaerobically by initial screening at 20 degrees Celsius using the sitting drop method on 96 Well MRC 2 Drop polystyrene Crystallization Plates SWISSCI in a Coy tent containing an N2/H2 (97:3%) atmosphere. The reservoir contained 90 ul of 25 % (w/v) glycerol ethoxylate 1,000, 100 mM Tris/HCl pH 8.5, and 100 mM CaCl2. 0.6 ul of crystallization solution was mixed with 0.6 ul of the protein, containing a final concentration of 2 mM acetyl-CoA. The protein was concentrated at 15 mg/ml in 25 mM Tris/HCl pH 7.6, 10% (v/v) glycerol, and 2 mM dithiothreitol. The crystals appeared in few weeks and were soaked in the crystallization solution supplemented with 15% (v/v) Ethylene glycol for a few seconds before freezing in liquid nitrogen.