8AF7

Room temperature SSX crystal structure of CTX-M-14 (10K dataset)

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	PETRA III, DESY BEAMLINE P11
Synchrotron site	PETRA III, DESY
Beamline	P11
Temperature [K]	295
Detector technology	PIXEL
Collection date	2018-12-16
Detector	DECTRIS PILATUS 6M
Wavelength(s)	1.0332
Spacegroup name	P 32 2 1
Unit cell lengths	42.160, 42.160, 234.350
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	16.540 - 1.550
R-factor	0.1805
Rwork	0.180
R-free	0.21470
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	6gth
RMSD bond length	0.006
RMSD bond angle	0.814
Data reduction software	CrystFEL
Data scaling software	CrystFEL
Phasing software	PHENIX
Refinement software	PHENIX (1.17.1_3660)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	17.770	1.580
High resolution limit [Å]	1.550	1.550
Number of reflections	36504	3514
<I/σ(I)>	3.25	0.62
Completeness [%]	100.0	100
Redundancy	114	65
CC(1/2)	0.815	0.268

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	BATCH MODE	4.5	293	CTX-M-14 was produced, purified and crystallized as before, with a slight modification in crystallization conditions to obtain slightly bigger crystals which roughly match the X-ray focal spot. For this purpose, a 50% CTX-M-14 solution (22 mg/ml) was mixed with a 45% precipitant solution (40% PEG8000, 200 mM lithium sulfate, 100 mM sodium acetate, pH 4.5) and with a 5% undiluted seed stock in batch crystallization setups, resulting in crystals with a homogeneous size distribution of 11-15 um after 90 minutes. The crystals were centrifuged at 200xg for 5 min and the supernatant was replaced with a stabilization buffer (28% PEG8000, 140 mM lithium sulfate, 70 mM sodium acetate, pH 4.5) to stop further crystal growth.