6CF3
Ethylene forming enzyme Y306A variant in complex with manganese and 2-oxoglutarate
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 EIGER X 9M |
Wavelength(s) | 0.9762 |
Spacegroup name | P 2 21 21 |
Unit cell lengths | 44.370, 85.410, 87.170 |
Unit cell angles | 90.00, 90.00, 90.00 |
Refinement procedure
Resolution | 44.370 - 1.120 |
R-factor | 0.1488 |
Rwork | 0.148 |
R-free | 0.16730 |
Structure solution method | MOLECULAR REPLACEMENT |
Starting model (for MR) | 5v2y |
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] | 44.370 | 44.370 | 1.140 |
High resolution limit [Å] | 1.120 | 6.130 | 1.120 |
Rmerge | 0.121 | 0.095 | 0.641 |
Rmeas | 0.133 | 0.105 | 0.725 |
Rpim | 0.053 | 0.044 | 0.329 |
Number of reflections | 125336 | 899 | 5884 |
<I/σ(I)> | 7.9 | ||
Completeness [%] | 98.2 | 99.1 | 94 |
Redundancy | 5.9 | 5.7 | 4.6 |
CC(1/2) | 0.992 | 0.982 | 0.707 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, SITTING DROP | 6.5 | 277.15 | 0.5 ul 64 mg/ml EFE-Y306A (1 mM manganese chloride, 25 mM HEPES pH 8.0, 1 mM TCEP, 3 mM L-Arginine, 3 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 five minutes in 25% w/v Ethylene glycol, 75% reservoir solution before freezing it. |