6G0K

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	79.842, 128.852, 236.250
Unit cell angles	90.00, 93.88, 90.00

Refinement procedure

Resolution	47.142 - 2.900
R-factor	0.2007
Rwork	0.198
R-free	0.24680
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.016
RMSD bond angle	1.400
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	AMoRE
Refinement software	PHENIX ((1.10.1_2155: ???))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	47.142	3.060
High resolution limit [Å]	2.900	2.900
Rmerge	0.159	0.793
Rmeas	0.186	0.930
Rpim	0.095	0.481
Number of reflections	52908	7679
<I/σ(I)>	8.5	2.5
Completeness [%]	100.0	100
Redundancy	3.8	3.8

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	EVAPORATION		292	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.