7R3I

PROSS optimitzed variant of RhlR (61 mutations) in complex with the synthetic antagonist mBTL

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	PETRA III, DESY BEAMLINE P11
Synchrotron site	PETRA III, DESY
Beamline	P11
Temperature [K]	100
Detector technology	PIXEL
Collection date	2020-10-22
Detector	DECTRIS PILATUS 6M-F
Wavelength(s)	1.0332
Spacegroup name	P 41 21 2
Unit cell lengths	85.904, 85.904, 202.593
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	43.630 - 3.100
R-factor	0.2492
Rwork	0.248
R-free	0.27240
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	structure prediction of RhlR with phyre2
Data reduction software	XDS
Data scaling software	Aimless (0.7.7)
Phasing software	PHASER
Refinement software	PHENIX (1.19.2)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	43.630	43.630	3.310
High resolution limit [Å]	3.100	8.770	3.100
Rmerge	0.129	0.048	0.846
Rmeas	0.131	0.050	0.862
Rpim	0.026	0.011	0.166
Total number of observations	365564	14467	68188
Number of reflections	14516	745	2557
<I/σ(I)>	21.9	52.7	5.3
Completeness [%]	100.0	99.4	100
Redundancy	25.2	19.4	26.7
CC(1/2)	0.999	0.999	0.944

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP		293	50 mM BICINE pH 8.8, 8.6 % (w/v) PEG 2K, 17.1 % (v/v) PEG MME 350 3 mg/ml 10 % (v/v) (2R,3R) -(-)-2,3-Butanediol