7RON

Crystal structure of the Friedel-Crafts alkylating enzyme CylK from Cylindospermum licheniforme

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 21-ID-F
Synchrotron site	APS
Beamline	21-ID-F
Temperature [K]	80
Detector technology	CCD
Collection date	2020-02-14
Detector	MARMOSAIC 300 mm CCD
Wavelength(s)	0.97872
Spacegroup name	C 2 2 21
Unit cell lengths	93.667, 138.613, 99.438
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	42.410 - 1.680
R-factor	0.1573
Rwork	0.156
R-free	0.18930
Structure solution method	SAD
RMSD bond length	0.016
RMSD bond angle	1.921
Data reduction software	HKL-2000
Data scaling software	SCALEPACK
Phasing software	PHASER
Refinement software	REFMAC (5.8.0258)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	50.000	50.000	1.710
High resolution limit [Å]	1.680	4.560	1.680
Rmerge	0.097	0.032	1.279
Rmeas	0.101	0.033	1.326
Rpim	0.026	0.009	0.346
Total number of observations	1088178
Number of reflections	73688	3915	3677
<I/σ(I)>	7.3
Completeness [%]	99.8	99.9	99.3
Redundancy	14.8	14.3	14.3
CC(1/2)		0.999	0.780

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7	298	Prior to crystallization trials, purified CylK protein in storage buffer (20 mM HEPES pH 7.8, 50 mM NaCl, 10 mM MgCl2, 10 mM CaCl2, 10% glycerol) was combined with 5-undecylbenzene-1,3-diol (UDR), a substrate analog dissolved in protein storage buffer containing 22% DMSO. The final protein solution contained 10 mg/mL CylK and 1.66 mM UDR. Crystals of undecylresorcinol-bound CylK were obtained by using the hanging drop vapor diffusion method in 2 uL drops mixed in 1:1 ratio with a precipitant solution of 1.8 M sodium malonate, pH 7.0. Crystals formed within about 1 week at room temperature