7NXG

Wax synthase 1 from Acinetobacter baylyi (AbWSD1) co-crystallized with myristic acid

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SLS BEAMLINE X10SA
Synchrotron site	SLS
Beamline	X10SA
Temperature [K]	100
Detector technology	PIXEL
Collection date	2014-05-05
Detector	DECTRIS PILATUS 6M
Wavelength(s)	0.9788
Spacegroup name	I 4 2 2
Unit cell lengths	118.320, 118.320, 142.560
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	44.100 - 1.950
R-factor	0.1896
Rwork	0.189
R-free	0.20930
Structure solution method	SAD
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	SHELXCD
Refinement software	PHENIX (1.19.1)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	44.100	50.000	2.050
High resolution limit [Å]	1.950	17.000	1.950
Rmerge	0.087	0.021	2.119
Rmeas	0.091	0.022	2.206
Total number of observations	487337
Number of reflections	36998	74	5049
<I/σ(I)>	19.91	78.77	1.46
Completeness [%]	99.7	97.4	99.4
Redundancy	13.172	10.095	12.699
CC(1/2)	1.000	1.000	0.659

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP		277.15	Native crystal: Crystals were grown by in situ proteolysis with trypsin. Trypsin was added in a 1:200 (w/w) ration to 20 mg/mL native His6-TF-AbWSD1 before set up of the crystallization screens. 100 nL precipitant (17.3 % ethanol, 0.2 M MgCl2, 0.1 M TRIS pH 8.0) were added to 200 nL protein using 96-well plates with a Cartesian pipetting robot (Zinsser Analytic). Plates were stored at 277.15 K. Crystals were transferred into a cryoprotectant consisting of mother liquor supplemented with 30 % ethylene glycol and then flash cooled in liquid nitrogen.
2	VAPOR DIFFUSION, SITTING DROP		277.15	SeMet crystal: Trypsin was added in a 1:1000 ratio (w/w) to 15 mg/mL selenomethionine-labelled His6-TF-AbWSD1. Sitting drops consisting of 100 nL protein and 100 nL precipitant (14.1 % ethanol, 0.1 M MgCl2, 0.1 M HEPES pH 7.5) were pipetted with a Gryphon robot (Art Robbins Instruments). The 96-well plates were stored at 277.15 K. For cryoprotection of the crystals, 2-methyl-2,4-pentanediol was added to the crystallization drop by transferring it five times with a 0.4 mm LithoLoop (Molecular Dimensions) at 277.15 K. Crystals were then harvested from the drop and flash-cooled in liquid nitrogen.