5LBX
Structure of the T175V Etr1p mutant in the trigonal form P312 in complex with NADP and crotonyl-CoA
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | SYNCHROTRON |
Source details | SLS BEAMLINE X10SA |
Synchrotron site | SLS |
Beamline | X10SA |
Temperature [K] | 100 |
Detector technology | PIXEL |
Collection date | 2015-12-01 |
Detector | DECTRIS PILATUS 6M-F |
Wavelength(s) | 1.73913 |
Spacegroup name | P 31 2 1 |
Unit cell lengths | 93.498, 93.498, 231.203 |
Unit cell angles | 90.00, 90.00, 120.00 |
Refinement procedure
Resolution | 46.749 - 2.500 |
R-factor | 0.1835 |
Rwork | 0.182 |
R-free | 0.21240 |
Structure solution method | MOLECULAR REPLACEMENT |
Starting model (for MR) | 4w99 |
RMSD bond length | 0.007 |
RMSD bond angle | 1.002 |
Data reduction software | XDS |
Data scaling software | SCALA |
Phasing software | PHASER |
Refinement software | PHENIX ((1.10.1_2155: ???)) |
Data quality characteristics
Overall | Outer shell | |
Low resolution limit [Å] | 47.040 | 2.640 |
High resolution limit [Å] | 2.500 | 2.500 |
Rmerge | 0.124 | 1.511 |
Number of reflections | 41476 | |
<I/σ(I)> | 12.2 | 1.5 |
Completeness [%] | 100.0 | 100 |
Redundancy | 8.6 | 7.7 |
CC(1/2) | 0.997 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, SITTING DROP | 6.4 | 298 | Etr1p was crystallized at 15 mg/mL with 5 mM NADP and 5 mM of crotonyl-CoA in the gel filtration buffer (100 mM NaCl, 20 mM Tris-HCl pH 7.9). Crystals appeared after several days in 2.0 M (NH4)2SO4, 100 mM ADA/NaOH pH 6.4 at room temperature. The crystals were soaked in a solution containing 30% (v/v) glycerol, 2.0 M (NH4)2SO4, 100 mM ADA pH 6.4 prior to freezing in liquid nitrogen to prevent water-crystal formation. |