6EXY

Neutron crystal structure of perdeuterated galectin-3C in complex with glycerol

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	MAX II BEAMLINE I911-3
Synchrotron site	MAX II
Beamline	I911-3
Temperature [K]	100
Detector technology	CCD
Collection date	2016-05-15
Detector	MARMOSAIC 225 mm CCD
Wavelength(s)	1.0000
Spacegroup name	P 21 21 21
Unit cell lengths	37.314, 58.352, 63.867
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	28.200 - 1.700
R-factor	0.1522
Rwork	0.150
R-free	0.18730
Structure solution method	FOURIER SYNTHESIS
Starting model (for MR)	3zsj
RMSD bond length	0.010
RMSD bond angle	1.735
Data reduction software	HKL-2000
Data scaling software	Aimless
Phasing software	PHENIX (1.13_2998)
Refinement software	PHENIX (1.13_2998)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	43.100	1.710	1.120
High resolution limit [Å]	1.100	1.650	1.100
Rmerge	0.068	0.499	1.259
Rpim	0.020		0.543
Number of reflections	54414	1544
<I/σ(I)>	20.2	1.6	1.4
Completeness [%]	95.0	90	91.5
Redundancy	12.3	2.6	7
CC(1/2)	1.000		0.726

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.5	293	12-15% PEG 4000 OR PEG 3000, 0.1M MGCL2, 0.015M BETA MERCAPTOETHANOL, 0.1M TRIS-DCL, PD 7.9, 0.4M NaSCN. All dissolved in D2O. Crystal grown in a 15 + 15 microlitre sitting drop that was first equilibrated for 1 week. A small crystal grown at 20-28% PEG was introduced. The drop was fed with fresh protein by adding 3-4 micro litres of protein with 10 mM lactose every 3-4 days for 3 months. Then the lactose was exchanged for glycerol by dialysis for at least one month against 10% glycerol (1.37 M), 24% PEG 4000 in the same buffer. For details see Manzoni et al. (2016).
1	VAPOR DIFFUSION, SITTING DROP	7.5	293	12-15% PEG 4000 OR PEG 3000, 0.1M MGCL2, 0.015M BETA MERCAPTOETHANOL, 0.1M TRIS-DCL, PD 7.9, 0.4M NaSCN. All dissolved in D2O. Crystal grown in a 15 + 15 microlitre sitting drop that was first equilibrated for 1 week. A small crystal grown at 20-28% PEG was introduced. The drop was fed with fresh protein by adding 3-4 micro litres of protein with 10 mM lactose every 3-4 days for 3 months. Then the lactose was exchanged for glycerol by dialysis for at least one month against 10% glycerol (1.37 M), 24% PEG 4000 in the same buffer. For details see Manzoni et al. (2016).