5EHU
sfGFP mutant with unnatural amino acid 4-azidoethoxy-L-phenylalanine incorporated at the 149 site
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | SYNCHROTRON |
Source details | APS BEAMLINE 24-ID-E |
Synchrotron site | APS |
Beamline | 24-ID-E |
Temperature [K] | 100 |
Detector technology | CCD |
Collection date | 2015-04-02 |
Detector | ADSC QUANTUM 315 |
Wavelength(s) | 0.979 |
Spacegroup name | C 1 2 1 |
Unit cell lengths | 129.917, 37.492, 91.848 |
Unit cell angles | 90.00, 106.31, 90.00 |
Refinement procedure
Resolution | 45.262 - 1.450 |
R-factor | 0.1756 |
Rwork | 0.175 |
R-free | 0.20830 |
Structure solution method | MOLECULAR REPLACEMENT |
Starting model (for MR) | 2b3p |
RMSD bond length | 0.009 |
RMSD bond angle | 1.357 |
Data reduction software | HKL-2000 |
Data scaling software | HKL-2000 |
Phasing software | PHASER |
Refinement software | PHENIX (1.9_1692) |
Data quality characteristics
Overall | Outer shell | |
Low resolution limit [Å] | 50.000 | 1.480 |
High resolution limit [Å] | 1.450 | 1.450 |
Rmerge | 0.089 | 0.385 |
Number of reflections | 73332 | |
<I/σ(I)> | 16.5 | 1.9 |
Completeness [%] | 96.4 | 94.3 |
Redundancy | 3.8 | 3.9 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, SITTING DROP | 7.5 | 298 | A 40 mg/mL solution of sfGFP-149-AePhe in a 20 mM Hepes buffer pH 7.5 was combined with a precipitation solution (20% PEG 8000, 100 mM Hepes pH 7.5) in a 1:1 ratio to form crystals in a sitting drop well at room temperature |