8ESL

Bile Salt Hydrolase from a Bacteroidales species with covalent inhibitor bound

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 23-ID-B
Synchrotron site	APS
Beamline	23-ID-B
Temperature [K]	100
Detector technology	PIXEL
Collection date	2022-02-27
Detector	DECTRIS EIGER X 16M
Wavelength(s)	1.00
Spacegroup name	P 21 21 21
Unit cell lengths	63.142, 66.812, 321.831
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	45.070 - 3.110
R-factor	0.2769
Rwork	0.277
R-free	0.30510
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	AlphaFold2 model of the protein
RMSD bond length	0.002
RMSD bond angle	0.410
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]	45.070	3.221
High resolution limit [Å]	3.110	3.110
Rmerge	0.240	1.710
Rmeas	0.287	2.057
Rpim	0.155	1.115
Number of reflections	24835	2378
<I/σ(I)>	3.81	0.74
Completeness [%]	94.8	80.39
Redundancy	3.4	3.3
CC(1/2)	0.978	0.121

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP		293	0.1 M HEPES: NaOH, pH 7.5, 25% (w/v) PEG 1000. 50 mL protein at 2.5 uM was incubated with 50 uM inhibitor for 1h at 37oC. Mixture was washed 3x with buffer in a spin concentrator and then concentrated to 6.77 mg/mL final concentration. Crystals formed in conditions with 1:2 ratio of protein:mother liquor.