6EZQ
human Serum Albumin complexed with NBD-C12 fatty acid
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | SYNCHROTRON |
Source details | SLS BEAMLINE X06DA |
Synchrotron site | SLS |
Beamline | X06DA |
Temperature [K] | 100 |
Detector technology | PIXEL |
Collection date | 2017-04-09 |
Detector | DECTRIS PILATUS 6M-F |
Wavelength(s) | 0.99992 |
Spacegroup name | C 1 2 1 |
Unit cell lengths | 98.480, 52.720, 123.300 |
Unit cell angles | 90.00, 110.10, 90.00 |
Refinement procedure
Resolution | 115.790 - 2.390 |
R-factor | 0.187 |
Rwork | 0.182 |
R-free | 0.29800 |
RMSD bond length | 0.010 |
RMSD bond angle | 1.260 |
Data reduction software | XDS ((VERSION Jun 1) |
Data scaling software | AutoPROC ((Version 1.1.7)) |
Phasing software | PHASER |
Refinement software | BUSTER (2.11.7) |
Data quality characteristics
Overall | Outer shell | |
Low resolution limit [Å] | 115.790 | 2.725 |
High resolution limit [Å] | 2.390 | 2.393 |
Rmerge | 0.084 | 0.620 |
Number of reflections | 13145 | |
<I/σ(I)> | 8.4 | 1.6 |
Completeness [%] | 55.3 | 8.5 |
Redundancy | 3.2 | 3.4 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, HANGING DROP | 7 | 292 | Essentially defatted human serum albumin from Sigma was purified by size exclusion chromatography to obtain pure monomeric protein. The purified HSA was dissolved in 50 mM potassium phosphate, 150 mM sodium chloride (pH 7.5) and concentrated to 2 mM (140 mg/mL). The HSA solution was incubated with a six fold excess of the NBD-labelled fatty acid at 4-5 deg.C for 4 hours. The final concentration of dimethyl sulfoxide was 2% (v/v). The crystal was grown by the hanging drop vapor diffusion method using a reservoir solution containing buffer (2.5 mM potassium phosphate, 7.5 mM sodium chloride, pH 7.0), 0.3% glycerol and polyethylene glycol 3350 (~30%). For crystallization 1 uL of HSA-ligand solution was equilibrated against 1 uL of reservoir solution. |