6CF3

Ethylene forming enzyme Y306A variant in complex with manganese and 2-oxoglutarate

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 21-ID-D
Synchrotron site	APS
Beamline	21-ID-D
Temperature [K]	100
Detector technology	PIXEL
Collection date	2017-06-03
Detector	DECTRIS EIGER X 9M
Wavelength(s)	0.9762
Spacegroup name	P 2 21 21
Unit cell lengths	44.370, 85.410, 87.170
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	44.370 - 1.120
R-factor	0.1488
Rwork	0.148
R-free	0.16730
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	5v2y
Data reduction software	MOSFLM
Data scaling software	Aimless (0.5.32)
Phasing software	PHASER (2.7.16)
Refinement software	PHENIX (1.13_2998)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	44.370	44.370	1.140
High resolution limit [Å]	1.120	6.130	1.120
Rmerge	0.121	0.095	0.641
Rmeas	0.133	0.105	0.725
Rpim	0.053	0.044	0.329
Number of reflections	125336	899	5884
<I/σ(I)>	7.9
Completeness [%]	98.2	99.1	94
Redundancy	5.9	5.7	4.6
CC(1/2)	0.992	0.982	0.707

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	6.5	277.15	0.5 ul 64 mg/ml EFE-Y306A (1 mM manganese chloride, 25 mM HEPES pH 8.0, 1 mM TCEP, 3 mM L-Arginine, 3 mM 2-oxoglutarate) was mixed with 0.5 ul reservoir solution. The sitting drop reservoir of 200 ul contained 25% w/v Polyethylene glycol 3,350, 0.1 M Bis-Tris pH 6.5, 0.2 M sodium chloride. The crystal was soaked for five minutes in 25% w/v Ethylene glycol, 75% reservoir solution before freezing it.