7R3G

PROSS optimitzed variant of RhlR (75 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	2020-03-13
Detector	DECTRIS PILATUS 6M-F
Wavelength(s)	1.0332
Spacegroup name	P 61
Unit cell lengths	164.780, 164.780, 40.311
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	47.570 - 2.150
R-factor	0.2009
Rwork	0.199
R-free	0.23010
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	structure prediction of phyre2
Data reduction software	XDS
Data scaling software	Aimless (0.7.4)
Phasing software	PHASER
Refinement software	PHENIX (1.19.2)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	47.570	47.570	2.220
High resolution limit [Å]	2.150	8.860	2.150
Rmerge	0.231	0.069	2.304
Rmeas	0.238	0.071	2.365
Rpim	0.053	0.017	0.529
Total number of observations	697687	9631	58554
Number of reflections	34628	553	2961
<I/σ(I)>	11.1	36.6	1.6
Completeness [%]	100.0	99.4	100
Redundancy	20.1	17.4	19.8
CC(1/2)	0.998	0.997	0.659

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP		293	0.1 M Bis-Tris pH 6.5, 0.2 M NaCl, 25 % (w/v) PEG 3350 3.5 mg/ml protein cryoprotectant: 10 % (v/v) (2R,3R) -(-)-2,3-Butanediol