4TVW

Resorufin ligase with bound resorufin-AMP analog

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 24-ID-C
Synchrotron site	APS
Beamline	24-ID-C
Temperature [K]	100
Detector technology	PIXEL
Collection date	2012-07-15
Detector	DECTRIS PILATUS 6M
Wavelength(s)	0.9795
Spacegroup name	P 1 21 1
Unit cell lengths	91.354, 99.932, 107.284
Unit cell angles	90.00, 105.79, 90.00

Refinement procedure

Resolution	49.966 - 3.505
R-factor	0.2032
Rwork	0.201
R-free	0.24840
RMSD bond length	0.003
RMSD bond angle	0.838
Refinement software	PHENIX ((phenix.refine: 1.9_1692))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	49.980	3.630
High resolution limit [Å]	3.500	3.500
Number of reflections	21542
<I/σ(I)>	7.8	2.333
Completeness [%]	92.0	66.7
Redundancy	2.8	1.9

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	6.5	277	2 uL of 5.6 mg/mL resorufin ligase containing 1 mM resorufin sulfamoyladenosine, 1 mM Mg(OAc)2, and 1 mM dithiothreitol mixed with 2 uL of precipitant solution (11% PEG 20,000, 0.15 M MES:NaOH, pH 6.5). Dark, and red colored crystalline plates appeared after ~ 5 days. Crystals were looped and washed through a cryoprotection solution of 80% precipitant solution (12% PEG 20,000, 0.2 M MES:NaOH, pH 6.5) and 20% glycerol. Crystals were then cryocooled by direction submersion into liquid nitrogen.