4RX2

A triple mutant in the omega-loop of TEM-1 beta-lactamase changes the substrate profile via a large conformational change and an altered general base for catalysis

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ALS BEAMLINE 5.0.1
Synchrotron site	ALS
Beamline	5.0.1
Temperature [K]	200
Detector technology	CCD
Collection date	2013-12-20
Detector	ADSC QUANTUM 315r
Wavelength(s)	0.997
Spacegroup name	P 1
Unit cell lengths	60.320, 83.300, 95.920
Unit cell angles	90.06, 89.97, 90.02

Refinement procedure

Resolution	62.890 - 2.315
R-factor	0.23667
Rwork	0.235
R-free	0.26795
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1btl
RMSD bond length	0.012
RMSD bond angle	1.519
Data reduction software	MOSFLM
Data scaling software	SCALA
Phasing software	MOLREP
Refinement software	REFMAC (5.8.0049)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	62.890	48.870	2.400
High resolution limit [Å]	2.315	8.980	2.315
Rmerge		0.099	0.481
Number of reflections	21750
<I/σ(I)>		7.5	3.1
Completeness [%]	75.0	99.6	99.3

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	6.5	298	0.1M HEPES pH6.5, 30% W/V PEG 6,000, VAPOR DIFFUSION, HANGING DROP, temperature 298K