7AP4

Thermus thermophilus Aspartyl-tRNA Synthetase in Complex with Compound AspS7HMDDA

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	82.277, 112.531, 88.253
Unit cell angles	90.00, 104.86, 90.00

Refinement procedure

Resolution	79.526 - 2.150
R-factor	0.1995
Rwork	0.199
R-free	0.24050
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1l0w
Data reduction software	XDS
Data scaling software	Aimless
Phasing software	PHASER
Refinement software	PHENIX (1.15.2_3472)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	79.530	79.530	2.170
High resolution limit [Å]	2.060	6.510	2.060
Rmerge	0.071	0.051	0.699
Rmeas	0.082	0.059	0.808
Rpim	0.041	0.030	0.401
Total number of observations	356281	11041	53328
Number of reflections	94888	3083	13811
<I/σ(I)>	9.5	21.4	1.7
Completeness [%]	99.0	98.7	98.6
Redundancy	3.8	3.6	3.9
CC(1/2)	0.997	0.996	0.741

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION	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.