7SXM

Structure of Xenon-derivatized Methyl-Coenzyme M Reductase from Methanothermobacter marburgensis

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ALS BEAMLINE 8.2.2
Synchrotron site	ALS
Beamline	8.2.2
Temperature [K]	100
Detector technology	CCD
Collection date	2017-04-07
Detector	ADSC QUANTUM 315r
Wavelength(s)	1.5498
Spacegroup name	P 1 21 1
Unit cell lengths	81.956, 115.741, 123.400
Unit cell angles	90.00, 92.53, 90.00

Refinement procedure

Resolution	69.630 - 2.503
R-factor	0.1803
Rwork	0.178
R-free	0.21800
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	3m1v
RMSD bond length	0.002
RMSD bond angle	0.531
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	PHENIX
Refinement software	PHENIX (1.18_3845)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	69.633	2.590
High resolution limit [Å]	2.503	2.503
Number of reflections	151627	14815
<I/σ(I)>	9.8
Completeness [%]	97.1
Redundancy	3
CC(1/2)		0.731

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP		293	1 uL of 24 mg/mL MCR was mixed with 1 uL of well solution (27% (w/v) PEG400, 0.18 M magnesium acetate, 0.25 M sodium chloride, and 0.10 M HEPES pH 7.5) to make a 2-uL sitting drop in a sealed well with 30 uL well solution. Yellowish green rod MCR crystals grew in two to four hrs. The entire crystallization plate was shipped to Advanced Light Source Beamline 8.2.2 for Xe- pressurization and data collection. The crystals used to determine Xe-derivatized structure were transferred from the sitting drop into 2-5 uL of paraffin oil briefly and then sealed in a steel chamber pressurized with xenon at 180 psi for 10 mins. The Xe-derivatized crystals were flash-cooled in liquid nitrogen for data collection immediately.