5V2Y

Ethylene forming enzyme in complex with manganese, 2-oxoglutarate and L-arginine

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 21-ID-G
Synchrotron site	APS
Beamline	21-ID-G
Temperature [K]	100
Detector technology	CCD
Collection date	2017-02-01
Detector	MARMOSAIC 300 mm CCD
Wavelength(s)	0.9786
Spacegroup name	P 21 21 21
Unit cell lengths	48.479, 82.116, 87.593
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	43.797 - 1.428
R-factor	0.1334
Rwork	0.132
R-free	0.16070
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	5v2t
RMSD bond length	0.007
RMSD bond angle	0.949
Data reduction software	XDS
Data scaling software	Aimless (0.5.31)
Phasing software	PHASER (2.7.16)
Refinement software	PHENIX (1.11.1-2575)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	43.800	43.800	1.450
High resolution limit [Å]	1.428	7.690	1.430
Rmerge	0.065	0.035	0.602
Rmeas	0.072	0.040	0.695
Rpim	0.029	0.018	0.341
Number of reflections	65410
<I/σ(I)>	15.9
Completeness [%]	99.7	96.9	96.3
Redundancy	5.8	4.8	3.9
CC(1/2)	0.999	0.998	0.773

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	6.5	277.15	0.2 ul 64 mg/ml EFE (+4 mM manganese chloride, 25 mM HEPES pH 8.0, 1 mM TCEP, 2.5 mM 2-oxoglutarate, 2.5 mM L-arginine) was mixed with 0.2 ul reservoir solution. The sitting drop reservoir of 50 ul contained 0.2 M sodium chloride, 0.1 M BIS-TRIS pH 6.5, 25% w/v Polyethylene glycol 3,350. The crystal was soaked for about a minute in 25% w/v Polyethylene glycol 400, 75% reservoir solution before freezing it.