5KMW

TOHO1 Beta lactamase mutant E166A/R274N/R276N -benzyl penicillin complex

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 19-ID
Synchrotron site	APS
Beamline	19-ID
Temperature [K]	15
Detector technology	CCD
Collection date	2015-05-01
Detector	ADSC QUANTUM 315r
Wavelength(s)	0.67
Spacegroup name	P 32 2 1
Unit cell lengths	72.262, 72.262, 98.193
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	10.000 - 1.100
R-factor	0.136
R-free	0.17000
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.015
Data scaling software	XDS (BUILT=20160617)
Phasing software	SHELX
Refinement software	SHELXL-97

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	38.620	1.160
High resolution limit [Å]	1.100	1.100
Rmerge	0.093	0.400
Number of reflections	118234
<I/σ(I)>	5.6	2.1
Completeness [%]	99.7	98.6
Redundancy	5.5	5.6

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	BATCH MODE	6.1	293.15	30 microliters of 10 mg/ml protein concentration was added to a solution containing 2.0 M ammonium sulfate and 0.1 M sodium citrate (pH 6.1). For ligand soaking, crystals were placed for 2-3 h in a reservoir solution containing 2.7 M ammonium sulfate, 0.1 M sodium citrate (pH 6.1), and 5.0 mM benzyl penicillin. The crystals were then placed momentarily in a reservoir solution containing a cryoprotectant (30% w/v trehalose) and subsequently flash-frozen in liquid nitrogen