5VKB

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

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	48.482, 81.743, 87.811
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	38.679 - 1.139
R-factor	0.1479
Rwork	0.147
R-free	0.16400
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	5v2z
RMSD bond length	0.008
RMSD bond angle	0.957
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]	48.480	1.160
High resolution limit [Å]	1.139	1.139
Rmerge	0.088	0.912
Rmeas	0.094	1.009
Rpim	0.032	0.422
Number of reflections	127532	5998
<I/σ(I)>	17.1
Completeness [%]	99.6	95.4
Redundancy	8.4	5.4
CC(1/2)	0.999	0.747

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 argininamide, 0.2 uL of 100 mM 2-oxoglutarate. 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 Polyethylene glycol 400, 75% reservoir solution before freezing it.