6EYM

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

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]	298
Detector technology	CCD
Collection date	2015-05-31
Detector	MARMOSAIC 325 mm CCD
Wavelength(s)	1.0000
Spacegroup name	P 21 21 21
Unit cell lengths	37.010, 58.360, 63.820
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	28.000 - 1.700
R-factor	0.17
Rwork	0.168
R-free	0.21100
Structure solution method	FOURIER SYNTHESIS
Starting model (for MR)	3zsj
RMSD bond length	0.011
RMSD bond angle	1.735
Data reduction software	LAUEGEN
Data scaling software	LSCALE
Phasing software	PHENIX
Refinement software	PHENIX ((1.12_2829: ???))

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	40.000	1.800	1.100
High resolution limit [Å]	1.070	1.700	1.070
Rmerge	0.032	0.334	0.817
Rpim		0.139
Number of reflections	61354	2172
<I/σ(I)>	24.7	3.9	2.1
Completeness [%]	99.4	87.9	96.8
Redundancy	6.6	4.9
CC(1/2)	1.000		0.772

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.5	295	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 drops that was first equilibrated for 1 week. A 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. For details see Manzoni et al. (2016).
1	VAPOR DIFFUSION, SITTING DROP	7.5	295	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 drops that was first equilibrated for 1 week. A 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. For details see Manzoni et al. (2016).
1	VAPOR DIFFUSION, SITTING DROP	7.5	295	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 drops that was first equilibrated for 1 week. A 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. For details see Manzoni et al. (2016).