7JWN

Crystal structure of Human Serum Albumin in complex with ketoprofen

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 19-ID
Synchrotron site	APS
Beamline	19-ID
Temperature [K]	100
Detector technology	PIXEL
Collection date	2020-02-11
Detector	DECTRIS PILATUS3 6M
Wavelength(s)	0.979
Spacegroup name	C 1 2 1
Unit cell lengths	170.499, 38.878, 98.501
Unit cell angles	90.00, 104.47, 90.00

Refinement procedure

Resolution	41.920 - 2.600
R-factor	0.1855
Rwork	0.183
R-free	0.23070
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4k2c
RMSD bond length	0.002
RMSD bond angle	1.139
Data reduction software	HKL-3000
Data scaling software	HKL-3000
Phasing software	MOLREP
Refinement software	REFMAC (5.8.0266)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	50.000	50.000	2.640
High resolution limit [Å]	2.600	7.050	2.600
Rmerge	0.080	0.062	0.803
Rmeas	0.092	0.071	0.925
Rpim	0.044	0.034	0.447
Number of reflections	18925	1022	851
<I/σ(I)>	9.3		1.3
Completeness [%]	96.4	96.3	88.5
Redundancy	4.2	4.2	3.5
CC(1/2)		0.990	0.612

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7	310	Prior to crystallization, the protein at concentration of 162 mg/ml (dissolved in 50 mM Tris pH 7.4 and 20 mM NaCl) was mixed with 100 mM ketoprofen in 100% DMSO in ratio 9:1 (final ketoprofen concentration 10 mM) and incubated for several hours at 37 oC. Then, 0.2 ul of the protein solution was mixed with 0.2 ul of the well condition (24% PEG 3350, 50 mM K2HPO4 at pH 7.0). The crystallization plate was incubated at RT for 3 months, then for several days at 37 oC, and after the growth of the first HSA crystals, the plate was transferred to RT.