6CBA

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

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 PILATUS3 S 6M
Wavelength(s)	0.9762
Spacegroup name	P 21 21 21
Unit cell lengths	48.430, 81.570, 87.630
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	32.491 - 1.130
R-factor	0.1426
Rwork	0.141
R-free	0.16290
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	5v2z
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]	48.430	48.430	1.150
High resolution limit [Å]	1.130	6.190	1.130
Rmerge	0.081	0.037	0.862
Rmeas	0.087	0.040	0.938
Rpim	0.032	0.015	0.362
Total number of observations	924312
Number of reflections	128283	914	6221
<I/σ(I)>	11.3
Completeness [%]	98.5	99.4	97.5
Redundancy	7.2	6.9	6.5
CC(1/2)	0.999	0.999	0.804

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 canavanine, 2 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 about a minute in 25% w/v Polyethylene glycol 400, 75% reservoir solution before freezing it.