6YXH

Cryogenic human alkaline ceramidase 3 (ACER3) at 2.6 A resolution determined by Serial Crystallography (SSX) using CrystalDirect

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SLS BEAMLINE X06SA
Synchrotron site	SLS
Beamline	X06SA
Temperature [K]	100
Detector technology	PIXEL
Collection date	2020-02-23
Detector	DECTRIS EIGER X 16M
Wavelength(s)	0.988
Spacegroup name	C 2 2 21
Unit cell lengths	61.470, 69.970, 259.490
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	46.180 - 2.600
R-factor	0.234
Rwork	0.232
R-free	0.28000
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	6g7o
RMSD bond length	0.010
RMSD bond angle	1.060
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	PHASER
Refinement software	BUSTER (2.10.3 (6-FEB-2020))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	50.000	2.620
High resolution limit [Å]	2.600	2.600
Number of reflections	17701	1275
<I/σ(I)>	8.4	0.98
Completeness [%]	100.0
Redundancy	36.01
CC(1/2)	0.990	0.330

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	LIPIDIC CUBIC PHASE		293	30 or 50 nl bolus overlaid with 600 nl precipitant solution in CrystalDirect plate-2. 41% PEG 400, 100 mM HEPES pH 7.5, 75 mM magnesium sulfate and 5% DMSO. Crystallisation experiments were carried out at the HTX facility of EMBL Grenoble. LCP bolus were harvested automatically at cryogenic condition using CrystalDirect technology.