8ESG

Bile Salt Hydrolase B from Lactobacillus gasseri 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]	100
Detector technology	PIXEL
Collection date	2021-08-01
Detector	DECTRIS PILATUS3 6M
Wavelength(s)	1.03
Spacegroup name	P 61 2 2
Unit cell lengths	157.362, 157.362, 140.718
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	48.950 - 2.180
R-factor	0.2366
Rwork	0.236
R-free	0.25510
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	7svh
RMSD bond length	0.002
RMSD bond angle	0.466
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]	48.950	2.258
High resolution limit [Å]	2.180	2.180
Rmerge	0.125	6.367
Rmeas	0.128	6.518
Rpim	0.028	1.380
Number of reflections	100941	5296
<I/σ(I)>	16.7	0.57
Completeness [%]	99.8	98.6
Redundancy	20.8	21.7
CC(1/2)	1.000	0.209

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP		293	0.2 M Lithium Sulfate, 0.1 M CAPS:NaOH, pH 10.5, 2M Ammonium Sulfate. Crystals formed in a 2:1 ratio of protein: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 8 mg/mL final concentration.