5V31

Ethylene forming enzyme in complex with manganese 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	48.630, 81.910, 87.160
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	43.580 - 2.450
R-factor	0.1828
Rwork	0.180
R-free	0.24680
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	5v2t
RMSD bond length	0.016
RMSD bond angle	0.915
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	Inner shell	Outer shell
Low resolution limit [Å]	87.160	87.160	2.550
High resolution limit [Å]	2.450	8.830	2.450
Rmerge	0.180	0.085	0.565
Rmeas	0.210	0.098	0.660
Rpim	0.106	0.049	0.334
Total number of observations	45124
Number of reflections	12891
<I/σ(I)>	5.4
Completeness [%]	97.3	91.7	98
Redundancy	3.5	3.7	3.6
CC(1/2)	0.972	0.984	0.718

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.5	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 L-arginine) was mixed with 0.2 ul reservoir solution. The sitting drop reservoir of 50 ul contained 0.1 M HEPES/ Sodium hydroxide pH 7.5, 25% w/v Polyethylene glycol 10,000. The crystal was soaked for about a minute in 25% w/v Polyethylene glycol 400, 75% reservoir solution before freezing it.