6G88

Crystal structure of Enterococcus Faecium D63r Penicillin-Binding protein 5 (PBP5fm)

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SOLEIL BEAMLINE PROXIMA 1
Synchrotron site	SOLEIL
Beamline	PROXIMA 1
Temperature [K]	100
Detector technology	PIXEL
Collection date	2013-06-10
Detector	DECTRIS PILATUS3 S 6M
Wavelength(s)	0.9801
Spacegroup name	C 1 2 1
Unit cell lengths	79.194, 128.736, 238.479
Unit cell angles	90.00, 94.04, 90.00

Refinement procedure

Resolution	47.577 - 3.300
R-factor	0.2029
Rwork	0.200
R-free	0.25060
RMSD bond length	0.004
RMSD bond angle	0.913
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	AMoRE
Refinement software	PHENIX ((1.13_2998: ???))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	47.580	3.480
High resolution limit [Å]	3.300	3.300
Rmerge	0.239	0.767
Rmeas	0.277	0.891
Rpim	0.140	0.449
Number of reflections	35868	5148
<I/σ(I)>	5.4
Completeness [%]	99.8	99.6
Redundancy	3.9	3.9

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	EVAPORATION		293	Crystals were grown by the sitting-drop vapor diffusion method. Crystals were grown at 20C by mixing 1microL of PBP5fm (48 mg/ml), 2microL of reservoir (20% PEG8000 (w/v), 0.4 M Li2SO4 and 100 mM Na acetate at pH 4.5) and 0.5 microL of 0.1M ceftobiprole (kind gift of Basilea Pharmaceutica. The crystal was cryoprotected in the same buffer containing 20 % (v/v) glycerol. Crystals were soaked for 5 min in 1 microl of 0.1 M ceftobiprole before cryoprotection.