6HHX

Structure of T. thermophilus AspRS in Complex with 5'-O-(N-(L-aspartyl)-sulfamoyl)cytidine

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE ID29
Synchrotron site	ESRF
Beamline	ID29
Temperature [K]	100
Detector technology	PIXEL
Collection date	2018-02-16
Detector	DECTRIS PILATUS 6M
Wavelength(s)	0.96863
Spacegroup name	P 1 21 1
Unit cell lengths	82.123, 112.706, 88.679
Unit cell angles	90.00, 104.40, 90.00

Refinement procedure

Resolution	57.780 - 2.100
R-factor	0.1888
Rwork	0.188
R-free	0.21780
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1l0w
Data reduction software	XDS
Data scaling software	Aimless (0.6.2)
Phasing software	PHASER
Refinement software	PHENIX (1.13_2998)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	68.320	68.320	2.140
High resolution limit [Å]	2.030	6.430	2.030
Rmerge	0.059	0.038	0.886
Rmeas	0.064	0.041	0.957
Rpim	0.025	0.016	0.360
Total number of observations	681569
Number of reflections	100487	3266	14665
<I/σ(I)>	14.9
Completeness [%]	100.0	99.7	100
Redundancy	6.8	6.4	7
CC(1/2)	0.999	0.999	0.767

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7	293	A 10 mg/ml protein solution was prepared in 10 mM TRIS pH 7.5, 100 mM NaCl, 2.5 mM DTT and 0.4% w/v low melting point agarose, maintaining the sample temperature at 315 kelvin. Crystals were grown by mixing an equal volume of the protein solution with 8-12% PEG 4000, 0.1 M Morpheus buffer system 1 (MES/imidazole) pH 7, 100 mM KCl, 20 v/v % glycerol. For soaking a 4 mM solution of compound in DMSO was used. A one third volume of the initial drop size was pipetted carefully onto the crystal containing drop. The sample was then placed back over the reservoir and incubated for approximately 2 hr. Crystals we caught in cryoloops and directly flash frozen in liquid nitrogen.