9SQT

Joint X-ray/neutron room temperature structure of perdeuterated LecA lectin in complex with deuterated galactose

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE BM07
Synchrotron site	ESRF
Beamline	BM07
Temperature [K]	291
Detector technology	PIXEL
Collection date	2024-10-07
Detector	DECTRIS PILATUS 6M
Wavelength(s)	0.92
Spacegroup name	P 1 21 1
Unit cell lengths	50.799, 106.522, 83.303
Unit cell angles	90.00, 94.63, 90.00

Refinement procedure

Resolution	45.900 - 1.900
Rwork	0.215
R-free	0.27800
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.014
RMSD bond angle	1.699
Data reduction software	XDS
Data scaling software	Aimless (0.7.15)
Phasing software	PHASER (2.8.3)
Refinement software	PHENIX (1.17.1)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	44.830	2.000	1.520
High resolution limit [Å]	1.490	1.900	1.490
Rmerge	0.090	0.423	1.688
Rmeas	0.111
Rpim		0.209	1.227
Number of reflections	142031	6352	7042
<I/σ(I)>	13	2.4	0.9
Completeness [%]	98.8	62.6	99.4
Redundancy	5.4		5.3
CC(1/2)	0.998		0.725

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	5	291	Buffer Composition (prepared in D2O instead of H2O):16% PEG 6000,1 M LiCl,0.1 M sodium acetate, pD 5 Initial Sitting Drop Setup: Mixed 20 uL of a protein solution containing: 9.6 mg/mL perdeuterated LecA, 5 mM Gal-d10, 5 uM calcium chloride (CaCl2),all dissolved in D2O. With 20 uL of the well solution. Crystal Growth & Feeding Process: Crystals initially appeared after 1 week. To promote further growth, the crystal was fed three times, each time with 5 uL of a pre-equilibrated drop prepared under the same conditions. Each feeding occurred one week apart, starting one week after initial crystal formation. This feeding strategy increased the crystal volume from 0.08 mm3 to 0.74 mm3.
1	VAPOR DIFFUSION, SITTING DROP	5	291	Buffer Composition (prepared in D2O instead of H2O):16% PEG 6000,1 M LiCl,0.1 M sodium acetate, pD 5 Initial Sitting Drop Setup: Mixed 20 uL of a protein solution containing: 9.6 mg/mL perdeuterated LecA, 5 mM Gal-d10, 5 uM calcium chloride (CaCl2),all dissolved in D2O. With 20 uL of the well solution. Crystal Growth & Feeding Process: Crystals initially appeared after 1 week. To promote further growth, the crystal was fed three times, each time with 5 uL of a pre-equilibrated drop prepared under the same conditions. Each feeding occurred one week apart, starting one week after initial crystal formation. This feeding strategy increased the crystal volume from 0.08 mm3 to 0.74 mm3.