8AVT

Racemic protein crystal structure of aureocin A53 from Staphylococcus aureus in the presence of glycerol 3-phosphate

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	DIAMOND BEAMLINE I03
Synchrotron site	Diamond
Beamline	I03
Temperature [K]	100
Detector technology	PIXEL
Collection date	2020-11-28
Detector	DECTRIS EIGER2 XE 16M
Wavelength(s)	0.8153
Spacegroup name	C 1 2 1
Unit cell lengths	73.718, 37.239, 114.088
Unit cell angles	90.00, 92.97, 90.00

Refinement procedure

Resolution	34.531 - 1.200
Rwork	0.176
R-free	0.20330
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2n8o
RMSD bond length	0.011
RMSD bond angle	1.703
Data reduction software	xia2
Data scaling software	Aimless
Phasing software	PHASER
Refinement software	REFMAC (5.8.0267)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	34.531	1.220
High resolution limit [Å]	1.200	1.200
Number of reflections	96953	4778
<I/σ(I)>	8.2	1.3
Completeness [%]	99.9	99.6
Redundancy	13.4	13.6
CC(1/2)	0.999	0.880

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	5.6	292	A racemic mixture of L-Aureocin A53 and D-Aureocin A53 at 40 mg/mL concentration was mixed 1:1 with 0.2 M ammonium acetate, 0.2 M sodium citrate and 29% PEG 4000 v/v precipitant condition, pH 5.6 in a 1 microlitre sitting drop. The crystal was soaked with sn-glycerol-3-phosphate (334 mM in the precipitant solution) at 1:1 v/v for 24 hours (50 equivalents). Crystals were submerged in 20% PEG 400 in the precipitant solution as cryoprotectant and flash frozen in liquid nitrogen during harvesting.