5VKA

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

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 21-ID-F
Synchrotron site	APS
Beamline	21-ID-F
Temperature [K]	100
Detector technology	CCD
Collection date	2017-04-16
Detector	MARMOSAIC 300 mm CCD
Wavelength(s)	0.9787
Spacegroup name	P 21 21 21
Unit cell lengths	48.583, 82.098, 87.766
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	38.701 - 1.169
R-factor	0.1732
Rwork	0.172
R-free	0.18760
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	5v2z
RMSD bond length	0.009
RMSD bond angle	0.961
Data reduction software	XDS
Data scaling software	Aimless (0.5.32)
Phasing software	PHASER (2.7.16)
Refinement software	PHENIX ((1.11.1_2575: ???))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	48.580	1.190
High resolution limit [Å]	1.169	1.169
Rmerge	0.094	0.593
Rmeas	0.103	0.687
Rpim	0.042	0.339
Number of reflections	118142	5146
<I/σ(I)>	12.3
Completeness [%]	99.2	87.9
Redundancy	5.7	3.6
CC(1/2)	0.997	0.752

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	8	277.15	0.5 ul 72 mg/ml selenomethionine containing EFE (9 mM manganese chloride, 25 mM HEPES pH 8.0, 1 mM TCEP) was mixed with 0.5 ul reservoir solution of 10 mM N-omega-hydroxy-L-arginine, and 0.2 uL of 100 mM 2OG. The sitting drop reservoir of 200 ul contained 20% w/v Polyethylene glycol 6,000, 0.1 M Tris pH 8.0, 0.2 M Lithium chloride. The crystal was soaked for about a minute in 25% w/v ethylene glycol, 75% reservoir solution before freezing it.