5LEJ

The Transcriptional Regulator PrfA from Listeria Monocytogenes in complex with a 30-bp operator PrfA-box motif

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE ID23-2
Synchrotron site	ESRF
Beamline	ID23-2
Temperature [K]	100
Detector technology	PIXEL
Collection date	2016-05-04
Detector	DECTRIS PILATUS 6M-F
Wavelength(s)	0.8729
Spacegroup name	P 43 21 2
Unit cell lengths	79.088, 79.088, 265.772
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	44.116 - 2.700
R-factor	0.2313
Rwork	0.230
R-free	0.26570
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2bgc
RMSD bond length	0.003
RMSD bond angle	0.511
Data reduction software	XDS
Data scaling software	Aimless
Phasing software	PHASER
Refinement software	PHENIX ((1.10.1_2155: ???))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	47.300	2.830
High resolution limit [Å]	2.700	2.700
Rmerge	0.089	1.184
Number of reflections	24171
<I/σ(I)>	16.8	1.9
Completeness [%]	99.8	99.4
Redundancy	8.6	8.6

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	5.5	291	Protein and duplex DNA were incubated together at a ratio of 1:1.3 (PrfA dimer:hly DNA) at final concentrations of 50 microM and 70 microM respectively in 20 mM Tris-HCl pH 8.0, 150 mM NaCl, 1 mM DTT for 60 min at room temperature, before being used for crystal setups. Crystals were obtained after 24 h by mixing 4 microL protein-DNA solution with 2 microL reservoir solution consisting of 8% PEG 8000, 100 mM sodium acetate pH 4.6, 100 mM magnesium acetate, 20% glycerol. Prior to vitrification the soaking of PrfAWT-DNA crystals were soaked in a reservoir solution containing 30% glycerol for 24 h.