9RUO

CdmB methyltransferase involved in the anaerobic dehalogenation of haloalkanes soaked with iodomethane

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE BM07
Synchrotron site	ESRF
Beamline	BM07
Temperature [K]	100
Detector technology	PIXEL
Collection date	2024-06-19
Detector	DECTRIS PILATUS 6M
Wavelength(s)	0.97951
Spacegroup name	C 1 2 1
Unit cell lengths	69.524, 86.152, 156.069
Unit cell angles	90.00, 99.27, 90.00

Refinement procedure

Resolution	30.920 - 1.620
R-factor	0.1789
Rwork	0.178
R-free	0.19890
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.007
RMSD bond angle	0.908
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]	77.020	1.790
High resolution limit [Å]	1.620	1.620
Rmerge	0.075	1.410
Rmeas	0.080	1.504
Rpim	0.028	0.514
Number of reflections	76703	3835
<I/σ(I)>	13	1.4
Completeness [%]	91.6	59.8
Redundancy	8	8.4
CC(1/2)	0.999	0.456

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	6.2	293.15	The protein was crystallized at 17.6 mg/ml in 25 mM Tris-HCl pH 7.6, 10% v/v glycerol, 0.1 mM ZnCl2, and 2 mM dithiothreitol. Initial screening was performed on a 96-Well MRC 2-Drop polystyrene Crystallization Plate (SWISSCI). From the initial hit, crystallization was further refined and performed aerobically at 20 degrees Celsius using the sitting drop method on CombiClover Jr crystallization plates (Molecular Dimensions). The reservoir chamber was filled with 100 ul of the crystallization solution containing: 50 % (v/v) Polyethylene glycol 200, 100 mM Sodium Potassium phosphate pH 6.2, and 200 mM Sodium chloride. The crystallization drop was formed by spotting 0.9 ul of purified protein with 0.9 ul of precipitant. The crystal was soaked in the crystallization solution supplemented with 16% (v/v) iodomethane (CH3I) for 4.8 minutes before being frozen in liquid nitrogen