5V2Z

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

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	2017-02-11
Detector	DECTRIS EIGER X 9M
Wavelength(s)	1.1272
Spacegroup name	P 21 21 21
Unit cell lengths	48.450, 81.860, 87.570
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	41.690 - 1.230
R-factor	0.142
Rwork	0.141
R-free	0.16000
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	5v2t
RMSD bond length	0.011
RMSD bond angle	1.110
Data reduction software	MOSFLM
Data scaling software	Aimless (0.5.31)
Phasing software	PHASER (2.7.16)
Refinement software	PHENIX (1.11.1-2575)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	48.450	1.250
High resolution limit [Å]	1.230	1.230
Rmerge	0.083	0.623
Rmeas		0.699
Rpim		0.311
Number of reflections	100282
<I/σ(I)>	14.6
Completeness [%]	98.7	84.1
Redundancy	10.1	4.8
CC(1/2)		0.816

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	6.5	277.15	0.5 ul 64 mg/ml EFE (+1 mM manganese chloride, 25 mM HEPES pH 8.0, 1 mM TCEP, +3 mM L-arginine, +3 mM 2-oxoadipic acid) was mixed with 0.2 ul reservoir solution. The sitting drop reservoir of 200 ul contained 25% PEG 3350, 100 mM Bis-Tris, pH 6.5, 200 mM NaCl. The crystal was soaked for about a minute in 25% w/v Polyethylene glycol 400, 75% reservoir solution before freezing it.