6T5I

The Transcriptional Regulator PrfA from Listeria Monocytogenes in complex with inhibitor of WNT production (IWP)-2

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	PETRA III, EMBL c/o DESY BEAMLINE P13 (MX1)
Synchrotron site	PETRA III, EMBL c/o DESY
Beamline	P13 (MX1)
Temperature [K]	100
Detector technology	PIXEL
Collection date	2019-09-20
Detector	DECTRIS PILATUS 6M
Wavelength(s)	0.9720
Spacegroup name	P 21 21 21
Unit cell lengths	47.951, 92.667, 101.447
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	46.334 - 2.000
R-factor	0.2012
Rwork	0.199
R-free	0.25230
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	6eut
Data reduction software	XDS
Data scaling software	Aimless
Phasing software	PHASER
Refinement software	PHENIX (1.14_3260)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	46.400	2.070
High resolution limit [Å]	2.000	2.000
Rmerge	0.135	2.078
Rpim	0.040	0.609
Number of reflections	30998	3033
<I/σ(I)>	12.6	1.2
Completeness [%]	99.2	98.8
Redundancy	12.9	13.1
CC(1/2)	0.999	0.435

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	5.5	291	Purified PrfA, in 200 mM NaCl and 20 mM NaP buffer at pH 6.5 was mixed with 1 mM IWP2 and 3.7 mM DTT prior to crystallization. Droplets of 1 microL protein solution at 3.1 mg ml-1 were mixed with 1 microL reservoir solution consisting of 29 % (w/v) PEG 4000, 100 mM sodium citrate pH 5.5, and 18 % (v/v) isopropanol. The dimethyl sulfoxide (DMSO) concentration was kept at 10% (v/v). Prior to vitrification, crystals were equilibrated for 2 days in a solution containing 35 % (w/v) PEG 4000, 100 mM sodium citrate pH 5.5 and 1 mM IWP-2.