6Y78

Structure of galectin-3C in complex with lactose determined by serial crystallography using a silicon nitride membrane support

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	MAX IV BEAMLINE BioMAX
Synchrotron site	MAX IV
Beamline	BioMAX
Temperature [K]	298
Detector technology	PIXEL
Collection date	2019-04-04
Detector	DECTRIS EIGER X 16M
Wavelength(s)	0.9800
Spacegroup name	P 21 21 21
Unit cell lengths	35.670, 56.360, 61.170
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	41.450 - 1.700
R-factor	0.1672
Rwork	0.166
R-free	0.19700
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	6eym
RMSD bond length	0.020
RMSD bond angle	1.593
Data reduction software	CrystFEL
Data scaling software	CrystFEL
Phasing software	PHENIX (1.16_3549)
Refinement software	PHENIX (1.17.1_3660)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	61.000	1.750
High resolution limit [Å]	1.700	1.700
Number of reflections	14154	1369
<I/σ(I)>	5.3	1.41
Completeness [%]	100.0	100
Redundancy	195.2	133.1

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7.5	277	Crystals were grown on XtalTool supports (Jena Bioscience) over a 24 well VDX-plate with 1 ml reservoir. The drop was made up of 1 microlitre 20 mg/ml protein in 10 mM sodium phosphate pH 7.4, 100 mM NaCl, 10 mM beta-mercaptoethanol, 2 mM lactose, 0.25 microlitres of galectin-3C seed crystals in a stabilization solution containing 0.1 M Tris pH 7.5, 33% (w/v) PEG 4000, 0.1 M MgCl2, 0.2 M NaSCN and 0.75 microlitres reservoir solution containing 0.1 M Tris pH 7.5, 34% (w/v) PEG 4000, 0.2 M NaSCN and 8 mM beta-mercapthoethanol. Crystals grew to a size of approximately 10-15 micrometer.