7OT1

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

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE MASSIF-3
Synchrotron site	ESRF
Beamline	MASSIF-3
Temperature [K]	100
Detector technology	PIXEL
Collection date	2018-12-03
Detector	DECTRIS EIGER X 4M
Wavelength(s)	0.967700
Spacegroup name	P 1 21 1
Unit cell lengths	70.273, 88.968, 84.749
Unit cell angles	90.00, 110.34, 90.00

Refinement procedure

Resolution	65.890 - 2.710
R-factor	0.2169
Rwork	0.214
R-free	0.28320
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]	65.890	65.890	2.860
High resolution limit [Å]	2.710	8.570	2.710
Rmerge	0.129	0.045	0.777
Rmeas	0.151	0.052	0.899
Rpim	0.078	0.027	0.447
Total number of observations	95934	3024	14991
Number of reflections	26108	869	3865
<I/σ(I)>	8.3	18.1	2.3
Completeness [%]	97.9	98.2	99.5
Redundancy	3.7	3.5	3.9
CC(1/2)	0.992	0.996	0.707

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.