7ZQX

Human galectin-3c in complex with a galactose derivative

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	2019-04-26
Detector	DECTRIS PILATUS 2M-F
Wavelength(s)	1
Spacegroup name	P 21 21 21
Unit cell lengths	36.404, 58.088, 62.575
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	29.060 - 1.050
R-factor	0.1097
Rwork	0.109
R-free	0.12910
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	5oax
RMSD bond length	0.020
RMSD bond angle	2.449
Data reduction software	XDS
Data scaling software	XDS
Phasing software	PHASER
Refinement software	REFMAC (5.8.0238)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	29.060	1.070
High resolution limit [Å]	1.050	1.050
Rmerge	0.037	0.545
Number of reflections	383590	62710
<I/σ(I)>	21.1	3
Completeness [%]	100.0
Redundancy	6.1
CC(1/2)	1.000	0.874

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	LIQUID DIFFUSION	7.5	289	The compound was dissolved in 100% DMSO to 50 mM concentration (2.30 mg + 86.2 uL DMSO). 15 uL galectin-3C (20 mg/ml in 10 mM Na phosphate buffer pH 7.4, 100 mM NaCl, 10 mM b-mercaptoethanol) were mixed with 0.5 uL compound solution. The sample was then centrifuged at 13 500 rpm for 5 min. A seed bead solution (Hampton Research) was used made from crushed apo galectin-3C crystals. The 40-fold diluted seed solution (diluted with reservoir solution similar to described below) was used to seed the crystallisation drops. The drops were set up in MRC 3-well plates and stored at 16C as follows: 200 nL galectin-3C complex + 30 nL app galectin-3C seed stock + 200 nL reservoir solution and equilibrated against 40 uL of reservoir containing 30% PEG 4000, 0.1 M Tris/HCl pH 7.5, 0.1 M MgCl2, 7.9 mM b- mercaptoethanol. Large crystals (0.1 - 0.2 mm) were briefly soaked in a cryoprotectant solution with the following composition: 25.5 % PEG 4000 15 % glycerol 85 mM Tris/HCl pH 7.5 85 mM MgCl2 250 mM NaSCN 8 mM b-mercaptoethanol 1 mM compound After soaking the crystals were flash-cooled in liquid N2.