6SJC

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

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SOLEIL BEAMLINE PROXIMA 1
Synchrotron site	SOLEIL
Beamline	PROXIMA 1
Temperature [K]	100
Detector technology	PIXEL
Collection date	2019-02-15
Detector	DECTRIS EIGER X 16M
Wavelength(s)	0.978565
Spacegroup name	P 1 21 1
Unit cell lengths	82.089, 112.764, 88.073
Unit cell angles	90.00, 104.17, 90.00

Refinement procedure

Resolution	57.581 - 2.230
R-factor	0.188
Rwork	0.187
R-free	0.23240
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1low
Data reduction software	XDS
Data scaling software	Aimless (0.7.4)
Phasing software	PHASER
Refinement software	PHENIX (1.15.2_3472)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	79.590	79.590	2.270
High resolution limit [Å]	2.160	6.820	2.160
Rmerge	0.064	0.034	1.095
Rmeas	0.069	0.037	1.184
Rpim	0.026	0.014	0.448
Total number of observations	583963	18595	83169
Number of reflections	83261	2720	12066
<I/σ(I)>	13.4	41.6	1.6
Completeness [%]	99.7	99.7	99.3
Redundancy	7	6.8	6.9
CC(1/2)	0.999	0.999	0.720

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 were caught in cryoloops and directly flash frozen in liquid nitrogen.