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.