7B1S

Crystal structure of the ethyl-coenzyme M reductase from Candidatus Ethanoperedens thermophilum at 0.994-A resolution

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	2019-10-08
Detector	DECTRIS PILATUS 2M-F
Wavelength(s)	1.00003
Spacegroup name	P 1 21 1
Unit cell lengths	83.736, 146.927, 113.128
Unit cell angles	90.00, 106.98, 90.00

Refinement procedure

Resolution	39.760 - 0.992
R-factor	0.1127
Rwork	0.112
R-free	0.12750
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	5a8w
Data reduction software	XDS
Data scaling software	STARANISO
Phasing software	PHASER
Refinement software	PHENIX (1.17.1_3660)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	108.190	1.070
High resolution limit [Å]	0.992	0.992
Rmerge	0.116	1.167
Rmeas	0.125	1.262
Rpim	0.047	0.476
Number of reflections	1061996	53103
<I/σ(I)>	9.7	1.6
Completeness [%]	89.1	58.1
Redundancy	7
CC(1/2)	0.997	0.563

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	8.5	293.15	Crystals were obtained by initial screening at 20 degree celsius using the sitting drop method on a 96-Well MRC 2-Drop Crystallization Plates in polystyrene (SWISSCI). The crystallization reservoir contained 90 ul of mother liquor, crystallization drop contained a mixture of 0.6 ul protein at 16.22 mg.ml-1 and 0.6 ul precipitant. The crystal was obtained by initial screening using the JBScreen Pentaerythritol screen from Jena Bioscience in a Coy tent under an N2/H2 atmosphere (95:5 %). The crystallization reservoir contained 45 % (w/v) Pentaerythritol Propoxylate (5/4 PO/OH), 100 mM Tris pH 8.5 and 400 mM potassium chloride.