6NIE

BesD with Fe(II), chloride, and alpha-ketoglutarate

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ALS BEAMLINE 8.3.1
Synchrotron site	ALS
Beamline	8.3.1
Temperature [K]	194
Detector technology	PIXEL
Collection date	2018-05-20
Detector	DECTRIS PILATUS3 S 6M
Wavelength(s)	1.11
Spacegroup name	P 1
Unit cell lengths	61.358, 68.916, 79.702
Unit cell angles	115.36, 91.57, 112.95

Refinement procedure

Resolution	70.027 - 1.950
R-factor	0.1791
Rwork	0.177
R-free	0.21590
Structure solution method	SAD
RMSD bond length	0.010
RMSD bond angle	0.982
Data reduction software	XDS
Data scaling software	Aimless
Phasing software	CRANK2
Refinement software	PHENIX ((1.13_2998: ???))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	70.030	2.020
High resolution limit [Å]	1.950	1.950
Number of reflections	149915	7341
<I/σ(I)>	10.7
Completeness [%]	97.3	95.7
Redundancy	13.6
CC(1/2)	0.998	0.780

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP		277	1:1 volume of protein solution (10 mg/mL BesD, lysine (1.5 mM), alpha-ketoglutarate (3 mM, pH 7)) and reservoir solution (MES pH 6.5 (100 mM), sodium chloride (0.6 mM), containing 20% (v/v) PEG 4000. Soaked anaerobically in a solution containing (NH4)2Fe(SO4)2 * 6H2O (1mM), alpha-ketoglutarate (1 mM), sodium ascorbate (1 mM), lysine (1 mM), and glycerol (20% (v/v) for 30 min before flash freezing in liquid nitrogen