8UGM

FphE, Staphylococcus aureus fluorophosphonate-binding serine hydrolases E, boronic acid-based compound Z27 bound

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	AUSTRALIAN SYNCHROTRON BEAMLINE MX2
Synchrotron site	Australian Synchrotron
Beamline	MX2
Temperature [K]	100
Detector technology	PIXEL
Collection date	2022-12-01
Detector	DECTRIS EIGER X 16M
Wavelength(s)	0.9537
Spacegroup name	P 1 21 1
Unit cell lengths	46.723, 73.971, 73.183
Unit cell angles	90.00, 91.43, 90.00

Refinement procedure

Resolution	39.820 - 1.650
R-factor	0.1516
Rwork	0.150
R-free	0.18930
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.010
RMSD bond angle	1.128
Data reduction software	XDS (20220220)
Data scaling software	Aimless (0.7.8)
Phasing software	PHASER (2.8.3)
Refinement software	PHENIX (1.20.1_4487)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	46.710	1.680
High resolution limit [Å]	1.650	1.650
Rmerge	0.112	0.944
Rmeas	0.123	1.072
Rpim	0.050	0.494
Total number of observations	350545	11116
Number of reflections	59687	2508
<I/σ(I)>	9.4	1.7
Completeness [%]	99.1
Redundancy	5.9	4.4
CC(1/2)	0.996	0.536

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	6.5	289.15	20uL 25.0 mg/mL FphE (10mM HEPES pH 7.5, 100mM NaCl) were mixed with 3uL Z27 (50mM in DMSO) and incubated at 4C overnight. 0.3 uL FphE-Z27 solution was mixed with 0.15 uL of reservoir solution. Sitting drop reservoir contained 25 uL of 180mM Calcium acetate, 100mM MES pH 6.5, 22.5% PEG 2000 MME. Crystal appeared within 2.5 days at 16C and grew until 14.5 days when it was frozen in a solution of ~25% glycerol, 75% reservoir.