5V2V

Ethylene forming enzyme in complex with nickel

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	2016-12-14
Detector	DECTRIS EIGER X 9M
Wavelength(s)	1.4777
Spacegroup name	P 2 21 21
Unit cell lengths	41.790, 79.310, 87.750
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	43.875 - 3.040
R-factor	0.2086
Rwork	0.199
R-free	0.26750
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	5v2t
RMSD bond length	0.005
RMSD bond angle	0.533
Data reduction software	iMOSFLM
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]	87.750	87.750	3.220
High resolution limit [Å]	3.040	9.120	3.040
Rmerge	0.141	0.069	0.382
Rmeas	0.157	0.078	0.424
Rpim	0.069	0.036	0.181
Total number of observations	28385
Number of reflections	5959
<I/σ(I)>	7
Completeness [%]	99.3	99.4	99.7
Redundancy	4.8	4.6	5.1
CC(1/2)	0.990	0.990	0.941

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	8	277.15	0.5 ul 72 mg/ml EFE (25 mM HEPES pH 8.0, 1 mM TCEP) was mixed with 0.5 ul reservoir solution. The sitting drop reservoir of 200 ul contained 0.2 M lithium chloride, 0.1 M Tris-HCl pH 8.0, 20% w/v Polyethylene glycol 6,000. A grown crystal was soaked for 2 hours in a sitting drop containing 0.1 M nickel chloride, 0.2 M lithium chloride, 0.1 M Tris-HCl pH 8.0, 20% w/v Polyethylene glycol 6,000, the reservoir of 100 ul contained 0.2 M lithium chloride, 0.1 M Tris-HCl pH 8.0, 20% w/v Polyethylene glycol 6,000.The crystal was direcly frozen.