7MBL

Crystal structure of Equine Serum Albumin in complex with Cobalt (II)

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 19-BM
Synchrotron site	APS
Beamline	19-BM
Temperature [K]	100
Detector technology	CCD
Collection date	2017-12-01
Detector	ADSC QUANTUM 315r
Wavelength(s)	0.97929
Spacegroup name	P 61
Unit cell lengths	93.312, 93.312, 141.510
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	46.700 - 2.700
R-factor	0.2174
Rwork	0.213
R-free	0.28920
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	5iiu
RMSD bond length	0.005
RMSD bond angle	1.342
Data reduction software	HKL-3000
Data scaling software	HKL-3000
Phasing software	HKL-3000
Refinement software	REFMAC (5.8.0267)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	50.000	50.000	2.750
High resolution limit [Å]	2.700	7.320	2.700
Rmerge	0.089	0.027	1.124
Rmeas	0.099	0.031	1.256
Rpim	0.044	0.014	0.557
Number of reflections	19239	980	982
<I/σ(I)>	7.1		1.3
Completeness [%]	99.9	98.7	100
Redundancy	5	4.6	5.1
CC(1/2)	0.991	0.998	0.751

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	COUNTER-DIFFUSION	7.4	291	Protein: 1 uL of 34 mg/mL ESA dissolved in 10 mM Tris (pH 7.4) and 150 mM NaCl. Precipitant: 1 uL of 0.2 M lithium sulfate, 2.0 M ammonium sulfate, 0.1 M Tris pH 7.4. 15-Well hanging drop crystallization plate (Qiagen, EasyXtal). 3.3 uL of 50 mM cobalt (II) chloride dissolved in the reservoir solution were added directly to the 2 uL crystallization drop containing crystals to reach a final cobalt concentration of 31 mM and then incubated for a few hours before harvesting. Paratone was used as a cryoprotectant