7OT3

Human Prolyl-tRNA Synthetase in Complex with L-proline and Compound 3b

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SLS BEAMLINE X06SA
Synchrotron site	SLS
Beamline	X06SA
Temperature [K]	100
Detector technology	PIXEL
Collection date	2021-04-22
Detector	DECTRIS EIGER X 16M
Wavelength(s)	1.000021
Spacegroup name	P 1 21 1
Unit cell lengths	71.871, 92.422, 87.260
Unit cell angles	90.00, 108.88, 90.00

Refinement procedure

Resolution	54.770 - 2.530
R-factor	0.2208
Rwork	0.218
R-free	0.27610
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	5vad
Data reduction software	XDS
Data scaling software	Aimless (0.7.4)
Phasing software	PHASER
Refinement software	PHENIX (1.17.1_3660)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	54.770	54.770	2.610
High resolution limit [Å]	2.480	7.840	2.480
Rmerge	0.047	0.029	0.985
Rmeas	0.051	0.032	1.064
Rpim	0.019	0.012	0.398
Total number of observations	264382	8573	38617
Number of reflections	38157	1267	5511
<I/σ(I)>	17.1	46.4	1.7
Completeness [%]	99.2	99.7	99.2
Redundancy	6.9	6.8	7
CC(1/2)	0.999	0.999	0.749

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	MICROBATCH	7.5	293	The purified protein at 30 mg/mL in 10 mM Tris pH 7, 100 mM NaCl, 2.5 mM 2-mercaptoethanol was incubated with 10 mM L-proline, 2 mM compound and 12% (v/v) DMSO on ice for 1 hr. Crystals were grown in a Terasaki Microbatch plate by mixing 1 uL the pre-mix with 1 uL of reservoir solution containing 0.25-0.4 M SrCl2, 15-20% (v/v) PEG3350 and 100 mM HEPES pH 7.5. The drops were covered with paraffin oil. Crystals were flash frozen in liquid nitrogen directly from the plate.