5VKA
Ethylene forming enzyme in complex with manganese, 2-oxoglutarate and N-omega-hydroxy-L-arginine
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 | 48.583, 82.098, 87.766 |
Unit cell angles | 90.00, 90.00, 90.00 |
Refinement procedure
Resolution | 38.701 - 1.169 |
R-factor | 0.1732 |
Rwork | 0.172 |
R-free | 0.18760 |
Structure solution method | MOLECULAR REPLACEMENT |
Starting model (for MR) | 5v2z |
RMSD bond length | 0.009 |
RMSD bond angle | 0.961 |
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] | 48.580 | 1.190 |
High resolution limit [Å] | 1.169 | 1.169 |
Rmerge | 0.094 | 0.593 |
Rmeas | 0.103 | 0.687 |
Rpim | 0.042 | 0.339 |
Number of reflections | 118142 | 5146 |
<I/σ(I)> | 12.3 | |
Completeness [%] | 99.2 | 87.9 |
Redundancy | 5.7 | 3.6 |
CC(1/2) | 0.997 | 0.752 |
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 N-omega-hydroxy-L-arginine, and 0.2 uL of 100 mM 2OG. 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 ethylene glycol, 75% reservoir solution before freezing it. |