5N1Q

METHYL-COENZYME M REDUCTASE III FROM METHANOTHERMOCOCCUS THERMOLITHOTROPHICUS AT 1.9 A RESOLUTION

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SLS BEAMLINE X10SA
Synchrotron site	SLS
Beamline	X10SA
Temperature [K]	100
Detector technology	PIXEL
Collection date	2015-11-30
Detector	DECTRIS PILATUS 6M
Wavelength(s)	0.99979
Spacegroup name	P 1 21 1
Unit cell lengths	111.811, 77.224, 145.458
Unit cell angles	90.00, 107.01, 90.00

Refinement procedure

Resolution	45.630 - 1.900
R-factor	0.1704
Rwork	0.169
R-free	0.19020
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	MCR III from Methanotorris formicicus
RMSD bond length	0.009
RMSD bond angle	1.040
Data reduction software	XDS
Data scaling software	SCALA (3.6.22)
Phasing software	MOLREP (11.2.08)
Refinement software	BUSTER (2.10.1)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	46.360	2.000
High resolution limit [Å]	1.900	1.900
Rmerge	0.087	0.577
Rpim	0.053	0.346
Number of reflections	185011	26774
<I/σ(I)>	8.1	1.9
Completeness [%]	99.2	98.7
Redundancy	3.7	3.7
CC(1/2)	0.994	0.804

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.6	291.15	1 ul of MCR III from M. thermolithotrophicus with a concentration of 35 mg/ml was mixed with 1 ul of reservoir solution. Best crystals with a yellow brick morphology appeared after a few days in 19% (w/v) polyethylene glycol 3350 and 200 mM MgCl2 in the absence of buffer. The crystals were immersed in a solution containing 19% (w/v) PEG 3350 and 200 mM MgCl2, 30% glycerol (v/v) prior to freezing in liquid nitrogen.