5V32

Ethylene forming enzyme in complex with manganese and malic acid

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 21-ID-G
Synchrotron site	APS
Beamline	21-ID-G
Temperature [K]	100
Detector technology	CCD
Collection date	2017-02-01
Detector	MARMOSAIC 300 mm CCD
Wavelength(s)	0.9786
Spacegroup name	I 2 2 2
Unit cell lengths	79.391, 97.856, 97.971
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	38.332 - 1.486
R-factor	0.1435
Rwork	0.142
R-free	0.16940
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	5v2t
RMSD bond length	0.008
RMSD bond angle	0.911
Data reduction software	XDS
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]	38.350	38.350	1.510
High resolution limit [Å]	1.486	8.140	1.490
Rmerge	0.078	0.029	0.948
Rmeas	0.086	0.032	1.042
Rpim	0.035	0.014	0.426
Number of reflections	62625
<I/σ(I)>	15.7
Completeness [%]	99.5	94.1	91
Redundancy	5.7	4.5	5.4
CC(1/2)	0.999	0.999	0.844

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7	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 2-oxoglutarate) was mixed with 0.2 ul reservoir solution. The sitting drop reservoir of 50 ul contained 0.15 M DL-Malic acid pH 7.0, 20% w/v Polyethylene glycol 3,350. The crystal was soaked for about a minute in 25% w/v Polyethylene glycol 400, 75% reservoir solution before freezing it.