5TRX

Room temperature structure of an extradiol ring-cleaving dioxygenase from B.fuscum determined using serial femtosecond crystallography

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	FREE ELECTRON LASER
Source details	SLAC LCLS BEAMLINE CXI
Synchrotron site	SLAC LCLS
Beamline	CXI
Temperature [K]	293
Detector technology	PIXEL
Collection date	2015-06-27
Detector	CS-PAD CXI-2
Wavelength(s)	1.5498
Spacegroup name	P 21 21 2
Unit cell lengths	111.570, 154.860, 101.800
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	20.000 - 2.380
R-factor	0.1869
Rwork	0.184
R-free	0.23210
Structure solution method	FOURIER SYNTHESIS
Starting model (for MR)	3ojt
RMSD bond length	0.011
RMSD bond angle	1.358
Data reduction software	CrystFEL (0.6.2-984ba3a8)
Data scaling software	CrystFEL (0.6.2-984ba3a8)
Refinement software	REFMAC (5.8.0069)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	34.500	2.470
High resolution limit [Å]	2.380	2.380
Number of reflections	71326	6975
<I/σ(I)>	2.43	3.42
Completeness [%]	99.9	99.5
Redundancy	26.5	13
CC(1/2)	0.810	0.450

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	BATCH MODE	5.8	293	Precipitant added to enzyme solution at 1:1 ratio: 12-14% PEG6000, 0.1-0.14 M calcium chloride, and 0.1 M MES pH 5.8. Crystallization carried out in a tube to produce saturated slurry of micro crystals