8ESI

Bile Salt Hydrolase from B. longum with covalent inhibitor bound

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 23-ID-D
Synchrotron site	APS
Beamline	23-ID-D
Temperature [K]	293
Detector technology	PIXEL
Collection date	2021-10-08
Detector	DECTRIS PILATUS3 6M
Wavelength(s)	1.00
Spacegroup name	P 1 21 1
Unit cell lengths	93.072, 166.801, 103.077
Unit cell angles	90.00, 116.60, 90.00

Refinement procedure

Resolution	49.100 - 2.350
R-factor	0.2103
Rwork	0.210
R-free	0.24360
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2hf0
RMSD bond length	0.002
RMSD bond angle	0.500
Data reduction software	XDS
Data scaling software	XDS
Phasing software	PHASER
Refinement software	PHENIX (1.20_4459)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	49.100	2.430
High resolution limit [Å]	2.350	2.350
Rmerge	0.177	1.889
Rmeas	0.208	2.199
Rpim	0.108	1.112
Number of reflections	116293	11592
<I/σ(I)>	5.4	0.88
Completeness [%]	99.6	99.6
Redundancy	3.8	3.9
CC(1/2)	0.986	0.341

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP		293	0.2 M MgCl2, 0.1 M Tris:HCl pH 8.5, 16% (w/v) PEG 4000. Crystals formed in 1:2 ratio of protein (11.8 mg/mL) to mother liquor. 2.5 uM protein was incubated with 50 uM inhibitor for 1h at 37oC. Mixture was washed 3x with buffer in a spin concentrator and then concentrated to 11.8 mg/mL final concentration.