8SBQ

FphE, Staphylococcus aureus fluorophosphonate-binding serine hydrolases E, fluorophosphonate JB101 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	46.936, 74.574, 73.862
Unit cell angles	90.00, 91.53, 90.00

Refinement procedure

Resolution	40.090 - 1.500
R-factor	0.1672
Rwork	0.166
R-free	0.18910
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.006
RMSD bond angle	0.914
Data reduction software	XDS
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]	46.700	1.520
High resolution limit [Å]	1.490	1.490
Rmerge	0.144	1.190
Rmeas	0.150	1.262
Rpim	0.041	0.406
Total number of observations	1076479	30839
Number of reflections	81242	3577
<I/σ(I)>	8	1.2
Completeness [%]	99.4
Redundancy	13.3	8.6
CC(1/2)	0.997	0.643

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	8	289.15	20uL 25.0 mg/ml FphE (10mM HEPES pH 7.5, 100mM NaCl) were mixed with 3uL fluorophosphonate molecule (10mM in DMSO) and incubated at 4C overnight. 0.2 ul 21.7 mg/ml FphE-fluorophosphonate were mixed with 0.2 ul of reservoir solution. Sitting drop reservoir contained 25 ul of 180mM Magnesium chloride hexahydrate, 100mM Tris pH 8.0, 22.5% PEG 2000 MME. Crystal appeared within a day at 16C and grew until day 3. It was frozen in a solution of ~25% glycerol, 75% reservoir.