9RUI

Apo state of the CdmB methyltransferase involved in the anaerobic dehalogenation of haloalkanes

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	69.511, 85.792, 155.797
Unit cell angles	90.00, 99.05, 90.00

Refinement procedure

Resolution	27.880 - 1.570
R-factor	0.1744
Rwork	0.173
R-free	0.19530
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.008
RMSD bond angle	1.000
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]	153.860	1.720
High resolution limit [Å]	1.570	1.570
Rmerge	0.062	1.230
Rmeas	0.066	1.329
Rpim	0.025	0.498
Number of reflections	82269	4113
<I/σ(I)>	15.7	1.5
Completeness [%]	90.6	57.2
Redundancy	7	6.8
CC(1/2)	0.999	0.530

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.