8CNR

Hybrid Cluster Protein from the thermophilic methanogen Methanothermococcus thermolithotrophicus as isolated in a reduced state at 1.45-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	2020-02-25
Detector	DECTRIS PILATUS 2M-F
Wavelength(s)	1.23984
Spacegroup name	P 21 21 2
Unit cell lengths	97.557, 102.454, 58.896
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	51.060 - 1.450
R-factor	0.1341
Rwork	0.133
R-free	0.15240
Structure solution method	SAD
RMSD bond length	0.009
RMSD bond angle	1.288
Data reduction software	autoPROC
Data scaling software	autoPROC
Phasing software	SHELXDE
Refinement software	PHENIX (1.20.1-4487)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	51.061	1.476
High resolution limit [Å]	1.450	1.451
Rmerge	0.119	1.193
Rmeas	0.132	1.361
Rpim	0.056	0.642
Number of reflections	103697	4833
<I/σ(I)>	9.5	1.4
Completeness [%]	98.8	92.4
Redundancy	5.3
CC(1/2)	0.996	0.510

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.6	293.15	Crystallisation was performed anaerobically at 20 degree Celsius using the sitting drop method on 96-Well MRC 2-Drop polystyrene Crystallisation Plates (SWISSCI) in a Coy tent containing a N2/H2 (97:3%) atmosphere. The reservoir chamber was filled with 90 ul of crystallisation condition and the drop was formed by spotting 0.55 ul protein with 0.55 ul of 20% (w/v) PEG 3,350 and 200 mM Magnesium formate. The protein was crystallized at 9.9 mg/ml in 25 mM Tris/HCl pH 7.6, 2 mM dithiothreitol and 10% (v/v) glycerol. Densities in the electron density map suggest a contamination from another crystallisation condition spatially close and containing 30% (v/v) 2-Methyl-2,4-pentanediol, 20 mM Calcium chloride and 100 mM Sodium acetate, pH 4.6. Crystals were cryo-protected by soaking in 20% (w/v) PEG 3,350 and 200mM Magnesium formate supplemented with 20% v/v glycerol for a few seconds.